bm mscs  (Thermo Fisher)


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    Name:
    StemPro BM Mesenchymal Stem Cells
    Description:
    StemPro BM Mesenchymal Stem Cells are cryopreserved human bone marrow mesenchymal stem cells BM MSCs These Off the Shelf cells are manufactured to meet Good Manufacturing Practice GMP manufacturing standards in a California licensed facility and available for Research Use Only They are isolated and expanded in culture to passage 4 under a low oxygen ischemia tolerant it proprietary clinical scale manufacturing process using Bovine Growth Serum BGS supplemented media and cryopreserved in an animal origin free cryopreservation medium The cells are supplied at 1x106 and 5x106 viable cells vial A Certificate of Analysis is available for each lot which documents the quality control specifications test results and donor information StemPro BM Mesenchymal Stem Cells allow you to • Minimize assay variability with itMSCs manufactured in compliance with GMP manufacturing standards• Improve functionality studies with immature high potency MSCs• No observed tumorigenicity and toxicity in GLP compliant animal studies• Accommodate experiments requiring one large lot of MSCs• Minimize your chance of assay failure when using tested cells reagents and protocolsImmature High Potency Low Oxygen Cultured MSCsStemPro BM Mesenchymal Stem Cells are derived from human bone marrow aspirates from qualified traceable donors The cells are isolated cultured and expanded in compliance with GMP manufacturing standards in a California licensed facility using a proprietary clinical scale Reduced Oxygen Tension manufacturing process Cells manufactured in reduced oxygen tension environments result in higher cell yields of highly potent immature stem cells compared to cells expanded in normal oxygen culture conditions Proven Non toxic ProfilesCells have demonstrated to be non toxic and non tumorigenic in GLP animal studies which were performed according to FDA guidelines A large lot can be made available for those requiring assay standardization Protocols using tested reagents will be provided to help minimize your chance of assay failure when using reagents and cells that have not been evaluated to work together View our tested media and reagents High Post Cryopreservation ViabilityStemPro Mesenchymal Stem Cells are supplied in CryoStor cryopreservation medium at 1x106 and 5x106 viable cells vial This medium is animal origin free and shown to support high cell viabilities post thawing Controlled processes are used for cryopreservation handling and shipping to help minimize risk of adverse effects to the quality of the cells Made in USA by Stemedica for Thermo Fisher Scientific
    Catalog Number:
    A15652
    Price:
    None
    Category:
    Eukaryotic Cells
    Applications:
    Cell Culture|Mesenchymal Stem Cell Culture|Stem Cell Culture|Stem Cell Research
    Buy from Supplier


    Structured Review

    Thermo Fisher bm mscs
    Sdf-1 impact on bone marrow mesenchymal stem cells. (A) Western blotting analysis of CXCR4, CD9, and tubulin in the whole population of bone marrow-derived mesenchymal stem cells <t>(BM-MSCs)</t> as well as of CXCR4 + and CXCR4 – BM-MSCs fractions fractions. (B) Quantitative RT-PCR analysis of CXCR4 and CD9 <t>mRNA</t> in BM-MSCs in control and Sdf-1-treated BM-MSCs. (C) Western blotting analysis of CD9 and tubulin in control and Sdf-1-treated (Sdf-1) BM-MSCs. (D) Migration of BM-MSCs in Sdf-1 gradient. The number of cells that migrated from the inserts was counted. (E) Percent of hybrid myotubes formed in co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs. (F) Co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs (green, localisation of β-galactosidase; blue, nuclei). Bar = 50 μm. CXCR, CXC chemokine receptor. * P
    StemPro BM Mesenchymal Stem Cells are cryopreserved human bone marrow mesenchymal stem cells BM MSCs These Off the Shelf cells are manufactured to meet Good Manufacturing Practice GMP manufacturing standards in a California licensed facility and available for Research Use Only They are isolated and expanded in culture to passage 4 under a low oxygen ischemia tolerant it proprietary clinical scale manufacturing process using Bovine Growth Serum BGS supplemented media and cryopreserved in an animal origin free cryopreservation medium The cells are supplied at 1x106 and 5x106 viable cells vial A Certificate of Analysis is available for each lot which documents the quality control specifications test results and donor information StemPro BM Mesenchymal Stem Cells allow you to • Minimize assay variability with itMSCs manufactured in compliance with GMP manufacturing standards• Improve functionality studies with immature high potency MSCs• No observed tumorigenicity and toxicity in GLP compliant animal studies• Accommodate experiments requiring one large lot of MSCs• Minimize your chance of assay failure when using tested cells reagents and protocolsImmature High Potency Low Oxygen Cultured MSCsStemPro BM Mesenchymal Stem Cells are derived from human bone marrow aspirates from qualified traceable donors The cells are isolated cultured and expanded in compliance with GMP manufacturing standards in a California licensed facility using a proprietary clinical scale Reduced Oxygen Tension manufacturing process Cells manufactured in reduced oxygen tension environments result in higher cell yields of highly potent immature stem cells compared to cells expanded in normal oxygen culture conditions Proven Non toxic ProfilesCells have demonstrated to be non toxic and non tumorigenic in GLP animal studies which were performed according to FDA guidelines A large lot can be made available for those requiring assay standardization Protocols using tested reagents will be provided to help minimize your chance of assay failure when using reagents and cells that have not been evaluated to work together View our tested media and reagents High Post Cryopreservation ViabilityStemPro Mesenchymal Stem Cells are supplied in CryoStor cryopreservation medium at 1x106 and 5x106 viable cells vial This medium is animal origin free and shown to support high cell viabilities post thawing Controlled processes are used for cryopreservation handling and shipping to help minimize risk of adverse effects to the quality of the cells Made in USA by Stemedica for Thermo Fisher Scientific
    https://www.bioz.com/result/bm mscs/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bm mscs - by Bioz Stars, 2021-07
    99/100 stars

    Images

    1) Product Images from "Sdf-1 (CXCL12) induces CD9 expression in stem cells engaged in muscle regeneration"

    Article Title: Sdf-1 (CXCL12) induces CD9 expression in stem cells engaged in muscle regeneration

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-015-0041-1

    Sdf-1 impact on bone marrow mesenchymal stem cells. (A) Western blotting analysis of CXCR4, CD9, and tubulin in the whole population of bone marrow-derived mesenchymal stem cells (BM-MSCs) as well as of CXCR4 + and CXCR4 – BM-MSCs fractions fractions. (B) Quantitative RT-PCR analysis of CXCR4 and CD9 mRNA in BM-MSCs in control and Sdf-1-treated BM-MSCs. (C) Western blotting analysis of CD9 and tubulin in control and Sdf-1-treated (Sdf-1) BM-MSCs. (D) Migration of BM-MSCs in Sdf-1 gradient. The number of cells that migrated from the inserts was counted. (E) Percent of hybrid myotubes formed in co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs. (F) Co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs (green, localisation of β-galactosidase; blue, nuclei). Bar = 50 μm. CXCR, CXC chemokine receptor. * P
    Figure Legend Snippet: Sdf-1 impact on bone marrow mesenchymal stem cells. (A) Western blotting analysis of CXCR4, CD9, and tubulin in the whole population of bone marrow-derived mesenchymal stem cells (BM-MSCs) as well as of CXCR4 + and CXCR4 – BM-MSCs fractions fractions. (B) Quantitative RT-PCR analysis of CXCR4 and CD9 mRNA in BM-MSCs in control and Sdf-1-treated BM-MSCs. (C) Western blotting analysis of CD9 and tubulin in control and Sdf-1-treated (Sdf-1) BM-MSCs. (D) Migration of BM-MSCs in Sdf-1 gradient. The number of cells that migrated from the inserts was counted. (E) Percent of hybrid myotubes formed in co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs. (F) Co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs (green, localisation of β-galactosidase; blue, nuclei). Bar = 50 μm. CXCR, CXC chemokine receptor. * P

    Techniques Used: Western Blot, Derivative Assay, Quantitative RT-PCR, Migration, Co-Culture Assay

    2) Product Images from "Increased Migration of Human Mesenchymal Stromal Cells by Autocrine Motility Factor (AMF) Resulted in Enhanced Recruitment towards Hepatocellular Carcinoma"

    Article Title: Increased Migration of Human Mesenchymal Stromal Cells by Autocrine Motility Factor (AMF) Resulted in Enhanced Recruitment towards Hepatocellular Carcinoma

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095171

    rAMF increases the in vitro chemotaxis of MSCs towards HCC and their adhesion to endothelial cells. A) Pretreatment of BM-MSCs with 1 µg/ml rAMF (black bars) increases chemotaxis towards TCM derived from HuH7 or HC-PT-5 cells compared to untreated cells (white bars). B) Wound-healing assay of MSCs after pretreatment with rAMF or control (DMEM). Representative images were taken 24 hours after scratching. C) Adhesion to HMEC-1 endothelial cells was increased in BM-MSCs exposed to rAMF. D) Expression of AMF receptor (AMFR), GDP dissociation inhibitor 2 (GDI-1), caveolin-1 (CAV-1) and caveolin-2 (CAV-2) by qRT-PCR. *p
    Figure Legend Snippet: rAMF increases the in vitro chemotaxis of MSCs towards HCC and their adhesion to endothelial cells. A) Pretreatment of BM-MSCs with 1 µg/ml rAMF (black bars) increases chemotaxis towards TCM derived from HuH7 or HC-PT-5 cells compared to untreated cells (white bars). B) Wound-healing assay of MSCs after pretreatment with rAMF or control (DMEM). Representative images were taken 24 hours after scratching. C) Adhesion to HMEC-1 endothelial cells was increased in BM-MSCs exposed to rAMF. D) Expression of AMF receptor (AMFR), GDP dissociation inhibitor 2 (GDI-1), caveolin-1 (CAV-1) and caveolin-2 (CAV-2) by qRT-PCR. *p

    Techniques Used: In Vitro, Chemotaxis Assay, Derivative Assay, Wound Healing Assay, Expressing, Quantitative RT-PCR

    rAMF increases the in vivo migration and anchorage of MSCs to HCC tumors. BM-MSCs prestimulated with 1 µg/ml of rAMF were labeled with DiR and CMDiI cell trackers and IV injected in SC HuH7 tumor-bearing mice. After 3 days, tumors were removed and exposed to FI. A) Total FI was calculated by measuring the region of interest (ROI) for all the tissues isolated and the results were expressed as total radiant efficiency. ns, non significant. B) Representative tumor images of mice inoculated with rAMF-prestimulated BM-MSCs (MSC-rAMF) or unstimulated cells (MSCs). Images represent the average radiant efficiency. Region of interest (ROI) was calculated for the isolated tumor (C), liver (D), lung (E) and spleen (F) and the results were expressed as the average radiant efficiency. **p
    Figure Legend Snippet: rAMF increases the in vivo migration and anchorage of MSCs to HCC tumors. BM-MSCs prestimulated with 1 µg/ml of rAMF were labeled with DiR and CMDiI cell trackers and IV injected in SC HuH7 tumor-bearing mice. After 3 days, tumors were removed and exposed to FI. A) Total FI was calculated by measuring the region of interest (ROI) for all the tissues isolated and the results were expressed as total radiant efficiency. ns, non significant. B) Representative tumor images of mice inoculated with rAMF-prestimulated BM-MSCs (MSC-rAMF) or unstimulated cells (MSCs). Images represent the average radiant efficiency. Region of interest (ROI) was calculated for the isolated tumor (C), liver (D), lung (E) and spleen (F) and the results were expressed as the average radiant efficiency. **p

    Techniques Used: In Vivo, Migration, Labeling, Injection, Mouse Assay, Isolation

    AMF potently stimulates in vitro chemotaxis of MSCs from different sources. A) Detection of AMF (55 kDa) by western blot in CCM derived from HCC cells and TCM from ex vivo HCC SC tumors (upper panel). Colloidal Coomassie staining was performed as loading control (lower panel). MSC migration was analyzed with a Boyden chamber assay using rAMF as chemoattractant for BM-MSCs (B), HUCPVCs (C) or AT-MSCs (D). Results were expressed as percentage of control (DMEM) ±SEM. *p
    Figure Legend Snippet: AMF potently stimulates in vitro chemotaxis of MSCs from different sources. A) Detection of AMF (55 kDa) by western blot in CCM derived from HCC cells and TCM from ex vivo HCC SC tumors (upper panel). Colloidal Coomassie staining was performed as loading control (lower panel). MSC migration was analyzed with a Boyden chamber assay using rAMF as chemoattractant for BM-MSCs (B), HUCPVCs (C) or AT-MSCs (D). Results were expressed as percentage of control (DMEM) ±SEM. *p

    Techniques Used: In Vitro, Chemotaxis Assay, Western Blot, Derivative Assay, Ex Vivo, Staining, Migration, Boyden Chamber Assay

    MMP3 expression and MMP2 activity is induced in MSCs by rAMF. A) Analysis of MMP3 expression by qRT-PCR in BM-MSCs (black bars), HUCPVCs (white bars) or AT-MSCs (gray bars) stimulated with 1 µg/ml of rAMF. **p
    Figure Legend Snippet: MMP3 expression and MMP2 activity is induced in MSCs by rAMF. A) Analysis of MMP3 expression by qRT-PCR in BM-MSCs (black bars), HUCPVCs (white bars) or AT-MSCs (gray bars) stimulated with 1 µg/ml of rAMF. **p

    Techniques Used: Expressing, Activity Assay, Quantitative RT-PCR

    3) Product Images from "Adipose Stem Cells Display Higher Regenerative Capacities and More Adaptable Electro-Kinetic Properties Compared to Bone Marrow-Derived Mesenchymal Stromal Cells"

    Article Title: Adipose Stem Cells Display Higher Regenerative Capacities and More Adaptable Electro-Kinetic Properties Compared to Bone Marrow-Derived Mesenchymal Stromal Cells

    Journal: Scientific Reports

    doi: 10.1038/srep37801

    ASCs are more resistant to in vitro hypoxia apoptosis compared to BM-MSCs: BM-MSCs and ASCs were exposed to 1% O 2 for 24 hours and then measured for morphological changes and apoptotic resistance. ( A ) BM-MSCs showed significant changes in cell morphology as cells became pyknotic while ASCs maintained their elongated, spindle fibroblast-like phenotype. ( B ) Cellular apoptosis after hypoxia conditioning was measured by FACS analysis using Annexin-V and PI staining ( n = 3).
    Figure Legend Snippet: ASCs are more resistant to in vitro hypoxia apoptosis compared to BM-MSCs: BM-MSCs and ASCs were exposed to 1% O 2 for 24 hours and then measured for morphological changes and apoptotic resistance. ( A ) BM-MSCs showed significant changes in cell morphology as cells became pyknotic while ASCs maintained their elongated, spindle fibroblast-like phenotype. ( B ) Cellular apoptosis after hypoxia conditioning was measured by FACS analysis using Annexin-V and PI staining ( n = 3).

    Techniques Used: In Vitro, FACS, Staining

    ASCs are more resistant to oxidative stress-induced senescence than BM-MSCs: BM-MSCs and ASCs were exposed to oxidative stress by treating cells with a dose of 600 μM H 2 O 2 . ( A ) Control cells, ( B ) H 2 O 2 treated cells showing more than 90% of BM-MSCs positive for SA-β-gal, and ASCs negative for SA-β-gal. ( C ) Representative images are displayed and data are shown as mean ± S.D. (error bars) of counted SA-Gal positive cells from 5 microscopic fields of 4 independent replicates. *****p
    Figure Legend Snippet: ASCs are more resistant to oxidative stress-induced senescence than BM-MSCs: BM-MSCs and ASCs were exposed to oxidative stress by treating cells with a dose of 600 μM H 2 O 2 . ( A ) Control cells, ( B ) H 2 O 2 treated cells showing more than 90% of BM-MSCs positive for SA-β-gal, and ASCs negative for SA-β-gal. ( C ) Representative images are displayed and data are shown as mean ± S.D. (error bars) of counted SA-Gal positive cells from 5 microscopic fields of 4 independent replicates. *****p

    Techniques Used:

    ( A ) ASCs have a significantly higher telomerase activity than BM-MSCs: Quantification of telomerase activity in BM-MSCs and ASCs was assayed using a fluorescent PCR-based telomerase activity detection method, TRAP (Telomeric Repeat Amplification Protocol). Telomerase activity is shown as amole of product/mg protein. ( B ) mRNA expression levels in ASCs with reference to BM-MSCs: qPCR analysis showed a higher expression of Oct4, VEGF, FGF, and TGF-β in ASCs while MMPII expression was higher in BM-MSCs. Data are shown as mean ± S.D. (error bars). ****p
    Figure Legend Snippet: ( A ) ASCs have a significantly higher telomerase activity than BM-MSCs: Quantification of telomerase activity in BM-MSCs and ASCs was assayed using a fluorescent PCR-based telomerase activity detection method, TRAP (Telomeric Repeat Amplification Protocol). Telomerase activity is shown as amole of product/mg protein. ( B ) mRNA expression levels in ASCs with reference to BM-MSCs: qPCR analysis showed a higher expression of Oct4, VEGF, FGF, and TGF-β in ASCs while MMPII expression was higher in BM-MSCs. Data are shown as mean ± S.D. (error bars). ****p

    Techniques Used: Activity Assay, Polymerase Chain Reaction, Amplification, Expressing, Real-time Polymerase Chain Reaction

    Traveling wave velocity of ( A ) ASCs and ( B ) BM-MSCs at 8 MHz during different time points: Snap shots of ( C ) ASCs and ( D ) BM-MSCs at frequency 8 MHz, 10 Vpp square wave.
    Figure Legend Snippet: Traveling wave velocity of ( A ) ASCs and ( B ) BM-MSCs at 8 MHz during different time points: Snap shots of ( C ) ASCs and ( D ) BM-MSCs at frequency 8 MHz, 10 Vpp square wave.

    Techniques Used:

    Representative histological analysis of hind limb muscles: Gastrocnemius muscles were collected after 4 weeks of cell therapy. Tissue samples were stained with: ( a ) H E showing muscle degeneration in the ischemic control group and infiltration of lymphocytes (*) compared to normal looking muscles in the BM-MSCs and ASCs treated groups ( b ) Positive staining for-CD31, in transplanted mice, especially In the ASCs-transplanted group (c) CD34 expression is pronounced in the BM-MSC-transplanted group ( d ) Increased expression of VEGF especially in the ASC-treated group (e) Staining with anti-αSMA is more pronounced in the ASCs group ( f ) staining of both tissues with anti-MMP9. Quantitative evaluation of the expression levels of CD31 ( g ), CD34 ( h ) and αSMA ( i ) was evaluated by counting the number of positive cells in each group. Data are shown as mean ± S.D. (error bars). Scale bars, 200 μm.
    Figure Legend Snippet: Representative histological analysis of hind limb muscles: Gastrocnemius muscles were collected after 4 weeks of cell therapy. Tissue samples were stained with: ( a ) H E showing muscle degeneration in the ischemic control group and infiltration of lymphocytes (*) compared to normal looking muscles in the BM-MSCs and ASCs treated groups ( b ) Positive staining for-CD31, in transplanted mice, especially In the ASCs-transplanted group (c) CD34 expression is pronounced in the BM-MSC-transplanted group ( d ) Increased expression of VEGF especially in the ASC-treated group (e) Staining with anti-αSMA is more pronounced in the ASCs group ( f ) staining of both tissues with anti-MMP9. Quantitative evaluation of the expression levels of CD31 ( g ), CD34 ( h ) and αSMA ( i ) was evaluated by counting the number of positive cells in each group. Data are shown as mean ± S.D. (error bars). Scale bars, 200 μm.

    Techniques Used: Staining, Mouse Assay, Expressing

    ( A ) Velocity of ASCs and BM-MSCs versus frequency. Snap shots of rotation of ( B ) ASCs and ( C ) BM-MSCs at 8 MHz, 10 Vpp.
    Figure Legend Snippet: ( A ) Velocity of ASCs and BM-MSCs versus frequency. Snap shots of rotation of ( B ) ASCs and ( C ) BM-MSCs at 8 MHz, 10 Vpp.

    Techniques Used:

    4) Product Images from "Potential Use of Amniotic Membrane - Derived Scaffold for Cerebrospinal Fluid Applications"

    Article Title: Potential Use of Amniotic Membrane - Derived Scaffold for Cerebrospinal Fluid Applications

    Journal: International Journal of Molecular and Cellular Medicine

    doi: 10.22088/IJMCM.BUMS.7.2.91

    BM-MSCs cultured on decellularized AM. A: scanning electron microscopy micrograph; B: hematoxylin and eosin staining, Arrows show BM-MSCs-seeded on AM scaffold
    Figure Legend Snippet: BM-MSCs cultured on decellularized AM. A: scanning electron microscopy micrograph; B: hematoxylin and eosin staining, Arrows show BM-MSCs-seeded on AM scaffold

    Techniques Used: Cell Culture, Electron Microscopy, Staining

    5) Product Images from "Engineered Stem Cells Improve Neurogenic Bladder by Overexpressing SDF-1 in a Pelvic Nerve Injury Rat Model"

    Article Title: Engineered Stem Cells Improve Neurogenic Bladder by Overexpressing SDF-1 in a Pelvic Nerve Injury Rat Model

    Journal: Cell Transplantation

    doi: 10.1177/0963689720902466

    (A) Representative images of cystometry in each group. Sham control is the normal control group. NB means neurogenic bladder. ImMSCs/eSDF-1 + are high SDF-1-expressing engineered MSCs, and NB+imMSCs/eSDF-1 − are engineered MSCs transfected by empty vectors. (B) Mean pressure of the voiding contractions compared in each group. * P
    Figure Legend Snippet: (A) Representative images of cystometry in each group. Sham control is the normal control group. NB means neurogenic bladder. ImMSCs/eSDF-1 + are high SDF-1-expressing engineered MSCs, and NB+imMSCs/eSDF-1 − are engineered MSCs transfected by empty vectors. (B) Mean pressure of the voiding contractions compared in each group. * P

    Techniques Used: Expressing, Transfection

    6) Product Images from "A Member of the Nuclear Receptor Superfamily, Designated as NR2F2, Supports the Self-Renewal Capacity and Pluripotency of Human Bone Marrow-Derived Mesenchymal Stem Cells"

    Article Title: A Member of the Nuclear Receptor Superfamily, Designated as NR2F2, Supports the Self-Renewal Capacity and Pluripotency of Human Bone Marrow-Derived Mesenchymal Stem Cells

    Journal: Stem Cells International

    doi: 10.1155/2016/5687589

    Osteogenic and adipogenic differentiation of transfected BM-MSCs. (a) Osteogenic differentiation of transfected BM-MSCs was detected by Alizarin Red staining. Scale bar = 100 μ m. (b) Adipogenic differentiation of transfected BM-MSCs was demonstrated via Oil red O staining. Scale bar = 100 μ m. (c) Mean relative values (±SD) of ALPL , BSP , RUNX2 , LPL , and PPAR-γ mRNA expression in Day 7 and Day 14 of induction culture processes.
    Figure Legend Snippet: Osteogenic and adipogenic differentiation of transfected BM-MSCs. (a) Osteogenic differentiation of transfected BM-MSCs was detected by Alizarin Red staining. Scale bar = 100 μ m. (b) Adipogenic differentiation of transfected BM-MSCs was demonstrated via Oil red O staining. Scale bar = 100 μ m. (c) Mean relative values (±SD) of ALPL , BSP , RUNX2 , LPL , and PPAR-γ mRNA expression in Day 7 and Day 14 of induction culture processes.

    Techniques Used: Transfection, Staining, Expressing

    Senescence analysis of transfected BM-MSCs. (a) Representative SA- β -Gal activity in different groups. Scale bar = 100 μ m. (b) Percentage of SA- β -Gal positive cells was quantified in different groups. Data was presented as mean ± SD. (c) Mean relative values (±SD) of P21 and P16 mRNA expression in different groups. (d) The confirmation of P21 upregulation in knock-down group by western blot analysis.
    Figure Legend Snippet: Senescence analysis of transfected BM-MSCs. (a) Representative SA- β -Gal activity in different groups. Scale bar = 100 μ m. (b) Percentage of SA- β -Gal positive cells was quantified in different groups. Data was presented as mean ± SD. (c) Mean relative values (±SD) of P21 and P16 mRNA expression in different groups. (d) The confirmation of P21 upregulation in knock-down group by western blot analysis.

    Techniques Used: Transfection, Activity Assay, Expressing, Western Blot

    Characteristics of transfected BM-MSCs. (a) Representative morphology of transfected BM-MSCs in negative control group. (b) More than 90% of BM-MSCs expressed GFP in negative control group. (c) Representative morphology of transfected BM-MSCs in knock-down group. (d) More than 90% of BM-MSCs expressed GFP in knock-down group. (e) Representative flow cytometric characterization of cell surface markers expressed on transfected BM-MSCs. Isotypic controls were represented by black line. The red line represented the negative control group and the blue line represented the knock-down group. (f) The knock-down of NR2F2 was confirmed by western blot analysis.
    Figure Legend Snippet: Characteristics of transfected BM-MSCs. (a) Representative morphology of transfected BM-MSCs in negative control group. (b) More than 90% of BM-MSCs expressed GFP in negative control group. (c) Representative morphology of transfected BM-MSCs in knock-down group. (d) More than 90% of BM-MSCs expressed GFP in knock-down group. (e) Representative flow cytometric characterization of cell surface markers expressed on transfected BM-MSCs. Isotypic controls were represented by black line. The red line represented the negative control group and the blue line represented the knock-down group. (f) The knock-down of NR2F2 was confirmed by western blot analysis.

    Techniques Used: Transfection, Negative Control, Flow Cytometry, Western Blot

    Self-renewal of transfected BM-MSCs. (a) Representative CFU-F of transfected BM-MSCs in a 6-well plate. (b) Mean value (±SD) of CFU-F number in different groups. (c) Mean relative values (±SD) of NANOG , OCT4 , and SOX2 mRNA expression in different groups. (d) Mean value (±SD) of OD values in different groups detected by MTT assay.
    Figure Legend Snippet: Self-renewal of transfected BM-MSCs. (a) Representative CFU-F of transfected BM-MSCs in a 6-well plate. (b) Mean value (±SD) of CFU-F number in different groups. (c) Mean relative values (±SD) of NANOG , OCT4 , and SOX2 mRNA expression in different groups. (d) Mean value (±SD) of OD values in different groups detected by MTT assay.

    Techniques Used: Transfection, Expressing, MTT Assay

    Apoptosis analysis of transfected BM-MSCs. (a) Representative flow cytometric results of apoptosis analysis in different groups. (b) Percentage of early and late apoptosis cells in different groups. Data was presented as mean ± SD. (c) Western blot analysis of Caspase-3 in different groups. The 17 kD and 19 kD bands represented cleaved Caspase-3.
    Figure Legend Snippet: Apoptosis analysis of transfected BM-MSCs. (a) Representative flow cytometric results of apoptosis analysis in different groups. (b) Percentage of early and late apoptosis cells in different groups. Data was presented as mean ± SD. (c) Western blot analysis of Caspase-3 in different groups. The 17 kD and 19 kD bands represented cleaved Caspase-3.

    Techniques Used: Transfection, Flow Cytometry, Western Blot

    7) Product Images from "Targeting ectodysplasin promotor by CRISPR/dCas9-effector effectively induces the reprogramming of human bone marrow-derived mesenchymal stem cells into sweat gland-like cells"

    Article Title: Targeting ectodysplasin promotor by CRISPR/dCas9-effector effectively induces the reprogramming of human bone marrow-derived mesenchymal stem cells into sweat gland-like cells

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-017-0758-0

    The effect of Dox-induced dCas9-E BM-MSCs on wound healing in vivo. a The mice were treated with Dox-induced dCas9-E BM-MSCs after full thickness scald injuries. b Sirius Red staining and Masson staining were used to detect the fibrosis, and the arrangement of collagen fibers was more regular than in the sham-treated group. Immunostaining was performed for c Ki67 (green) and d CD31 (red) after treatment with Dox-induced dCas9-E BM-MSCs. Scale bars = 20 μm. DAPI 4’6-diamidino-2-phenylindole, sgRNA single-guide RNA
    Figure Legend Snippet: The effect of Dox-induced dCas9-E BM-MSCs on wound healing in vivo. a The mice were treated with Dox-induced dCas9-E BM-MSCs after full thickness scald injuries. b Sirius Red staining and Masson staining were used to detect the fibrosis, and the arrangement of collagen fibers was more regular than in the sham-treated group. Immunostaining was performed for c Ki67 (green) and d CD31 (red) after treatment with Dox-induced dCas9-E BM-MSCs. Scale bars = 20 μm. DAPI 4’6-diamidino-2-phenylindole, sgRNA single-guide RNA

    Techniques Used: In Vivo, Mouse Assay, Staining, Immunostaining

    Involvement of NF-κB pathways in dCas9-E-induced EDA expression. The cells transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h exhibited a significance difference in NF-κB downstream Sonic Hedgehog (Shh) and cyclin D1 mRNA ( a ) and protein ( b ) expression. Immunostaining was also performed to detect the activity of the NF-κB pathway. dCas9-E BM-MSCs induced more NF-κB translocated into the nuclei and then activated the expression of Shh and cyclin D1 ( c ). All data are normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and calibrated based on the BM-MSC group. *** p
    Figure Legend Snippet: Involvement of NF-κB pathways in dCas9-E-induced EDA expression. The cells transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h exhibited a significance difference in NF-κB downstream Sonic Hedgehog (Shh) and cyclin D1 mRNA ( a ) and protein ( b ) expression. Immunostaining was also performed to detect the activity of the NF-κB pathway. dCas9-E BM-MSCs induced more NF-κB translocated into the nuclei and then activated the expression of Shh and cyclin D1 ( c ). All data are normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and calibrated based on the BM-MSC group. *** p

    Techniques Used: Expressing, Transfection, Incubation, Immunostaining, Activity Assay

    The effect of dCas9-E BM-MSCs on the regeneration of sweat glands in vivo. a The saline-treated paw (Sham) showed a negative result for the perspiration test, whereas paws implanted with Dox-induced dCas9-E BM-MSCs showed a positive result (sgRNA2). b Hematoxylin and eosin staining was used to detect the duct structure of sweat gland tissue after injection with Dox-induced dCas9-E BM-MSCs. c The expression of carcinoembryonic antigen (CEA; red), cytokeratin 19 (CK19; green) and nucleus (DAPI) in BM-MSCs after implantation for 20 days. d The quantification of each paw (0.5 cm × 0.5 cm, treated with collagenase I for 1 h at 37 °C) was also conducted after treatment with Dox-induced dCas9-E BM-MSCs. ** p
    Figure Legend Snippet: The effect of dCas9-E BM-MSCs on the regeneration of sweat glands in vivo. a The saline-treated paw (Sham) showed a negative result for the perspiration test, whereas paws implanted with Dox-induced dCas9-E BM-MSCs showed a positive result (sgRNA2). b Hematoxylin and eosin staining was used to detect the duct structure of sweat gland tissue after injection with Dox-induced dCas9-E BM-MSCs. c The expression of carcinoembryonic antigen (CEA; red), cytokeratin 19 (CK19; green) and nucleus (DAPI) in BM-MSCs after implantation for 20 days. d The quantification of each paw (0.5 cm × 0.5 cm, treated with collagenase I for 1 h at 37 °C) was also conducted after treatment with Dox-induced dCas9-E BM-MSCs. ** p

    Techniques Used: In Vivo, Staining, Injection, Expressing

    dCas9-E expression in BM-MSCs. a Bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with pLKO.1-puro-U6 and dCas9-E, and stained with PE-labeled anti-HA and DAPI 72 h post-transfection. b The expression of designed dCas9-E nucleases. Scale bar = 50 μm. DAPI 4’6-diamidino-2-phenylindole, GAPDH glyceraldehyde-3-phosphate dehydrogenase, HA hemagglutinin, sgRNA single-guide RNA
    Figure Legend Snippet: dCas9-E expression in BM-MSCs. a Bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with pLKO.1-puro-U6 and dCas9-E, and stained with PE-labeled anti-HA and DAPI 72 h post-transfection. b The expression of designed dCas9-E nucleases. Scale bar = 50 μm. DAPI 4’6-diamidino-2-phenylindole, GAPDH glyceraldehyde-3-phosphate dehydrogenase, HA hemagglutinin, sgRNA single-guide RNA

    Techniques Used: Expressing, Derivative Assay, Transfection, Staining, Labeling

    Characteristics of dCas9-E BM-MSCs after Dox induction. a The sweat gland markers, carcinoembryonic antigen (CEA) and cytokeratin (CK)19, were detected by qRT-PCR after bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h. b Proteins were collected from BM-MSCs transfected with dCas9-E. The sweat glands biomarkers CEA, CK7, CK14, and CK19 were detected by Western blotting using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for calibration of sample loading. c Immunofluorescence staining was used to detect CEA (red) and CK19 (green) in BM-MSCs (scale bar = 50 μm). d Villi ultrastructure changes after dCas9-E BM-MSCs were treated with Dox (scale bar = 1 μm). ** p
    Figure Legend Snippet: Characteristics of dCas9-E BM-MSCs after Dox induction. a The sweat gland markers, carcinoembryonic antigen (CEA) and cytokeratin (CK)19, were detected by qRT-PCR after bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h. b Proteins were collected from BM-MSCs transfected with dCas9-E. The sweat glands biomarkers CEA, CK7, CK14, and CK19 were detected by Western blotting using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for calibration of sample loading. c Immunofluorescence staining was used to detect CEA (red) and CK19 (green) in BM-MSCs (scale bar = 50 μm). d Villi ultrastructure changes after dCas9-E BM-MSCs were treated with Dox (scale bar = 1 μm). ** p

    Techniques Used: Quantitative RT-PCR, Derivative Assay, Transfection, Incubation, Western Blot, Immunofluorescence, Staining

    dCas9-E mediated activation of EDA in BM-MSCs. The cells were harvested and total mRNA and protein were extracted. Ectodysplasin (EDA) gene and protein expression were identified by qPCR ( a ) and Western blotting ( b ) after doxycycline (Dox) induction for 48 h. c Fluorescence microscopy detection of EDA expression activated by dCas9-E. After supplementing with 2 μg/ml Dox for 48 h, enhanced expression of EDA (red) was detected in dCas9-E transfected bone marrow-derived mesenchymal stem cells (BM-MSCs). The data are expressed as the mean ± SD. *** p
    Figure Legend Snippet: dCas9-E mediated activation of EDA in BM-MSCs. The cells were harvested and total mRNA and protein were extracted. Ectodysplasin (EDA) gene and protein expression were identified by qPCR ( a ) and Western blotting ( b ) after doxycycline (Dox) induction for 48 h. c Fluorescence microscopy detection of EDA expression activated by dCas9-E. After supplementing with 2 μg/ml Dox for 48 h, enhanced expression of EDA (red) was detected in dCas9-E transfected bone marrow-derived mesenchymal stem cells (BM-MSCs). The data are expressed as the mean ± SD. *** p

    Techniques Used: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Fluorescence, Microscopy, Transfection, Derivative Assay

    8) Product Images from "miR-146a-5p circuitry uncouples cell proliferation and migration, but not differentiation, in human mesenchymal stem cells"

    Article Title: miR-146a-5p circuitry uncouples cell proliferation and migration, but not differentiation, in human mesenchymal stem cells

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkt666

    Knocking down CXCL12 and SIKE1 inhibits WJ-MSC migratory ability, and CXCL12 silencing increases miR-146a-5p levels. ( A and B ) CXCL12 and SIKE1 control MSC motility. A total of 5 × 10 4 WJ-MSCs were transduced with CXCL12 (A) or SIKE1 (B) shRNA and then assessed by Transwell migration assay. Migrated cells were counted after 12 h. Results are shown as mean ± SD from two duplicates. * P
    Figure Legend Snippet: Knocking down CXCL12 and SIKE1 inhibits WJ-MSC migratory ability, and CXCL12 silencing increases miR-146a-5p levels. ( A and B ) CXCL12 and SIKE1 control MSC motility. A total of 5 × 10 4 WJ-MSCs were transduced with CXCL12 (A) or SIKE1 (B) shRNA and then assessed by Transwell migration assay. Migrated cells were counted after 12 h. Results are shown as mean ± SD from two duplicates. * P

    Techniques Used: Transduction, shRNA, Transwell Migration Assay

    miR-146a-5p regulates MSC migration and proliferation. ( A and B ) miR-146a-5p suppresses MSC motility. WJ-MSCs transfected with siRNAs against GFP or miR-146a-5p (siControl or si146a; A), and BM-MSCs transduced with lentiviruses expressing miR-146a-5p or empty lentivirus vector control (B) were used in Transwell assays. miR-146a-5p levels were detected by RT-qPCR (left panels). Results are shown as mean ± SD from four experiments. * P
    Figure Legend Snippet: miR-146a-5p regulates MSC migration and proliferation. ( A and B ) miR-146a-5p suppresses MSC motility. WJ-MSCs transfected with siRNAs against GFP or miR-146a-5p (siControl or si146a; A), and BM-MSCs transduced with lentiviruses expressing miR-146a-5p or empty lentivirus vector control (B) were used in Transwell assays. miR-146a-5p levels were detected by RT-qPCR (left panels). Results are shown as mean ± SD from four experiments. * P

    Techniques Used: Migration, Transfection, Transduction, Expressing, Plasmid Preparation, Quantitative RT-PCR

    9) Product Images from "Non-viral Gene Delivery of Interleukin-1 Receptor Antagonist Using Collagen-Hydroxyapatite Scaffold Protects Rat BM-MSCs From IL-1β-Mediated Inhibition of Osteogenesis"

    Article Title: Non-viral Gene Delivery of Interleukin-1 Receptor Antagonist Using Collagen-Hydroxyapatite Scaffold Protects Rat BM-MSCs From IL-1β-Mediated Inhibition of Osteogenesis

    Journal: Frontiers in Bioengineering and Biotechnology

    doi: 10.3389/fbioe.2020.582012

    Assessment of IL-1Ra produced by rat BM-MSCs transfected with PEI-pIL-1Ra nanoparticles. Quantification of IL-1Ra produced by BM-MSCs, collected in culture media at 3, 7, 10, and 14 days post-transfection. (A) Data is normalized to μg of dsDNA present in cultures at each timepoint. (B) Data is expressed as pg/ml. *** and **** denotes p
    Figure Legend Snippet: Assessment of IL-1Ra produced by rat BM-MSCs transfected with PEI-pIL-1Ra nanoparticles. Quantification of IL-1Ra produced by BM-MSCs, collected in culture media at 3, 7, 10, and 14 days post-transfection. (A) Data is normalized to μg of dsDNA present in cultures at each timepoint. (B) Data is expressed as pg/ml. *** and **** denotes p

    Techniques Used: Produced, Transfection

    Protective effect of PEI-pIL-1Ra nanoparticles on IL-1β-mediated inhibition of osteogenesis in BM-MSCs. Representative 3D microCT reconstructions depicting mineral deposition by BM-MSCs after 21 days in CHA scaffolds cultured under (A) basal medium, (B) osteogenic medium, (C) osteogenic medium + IL-1β (1 ng/ml), and (D) osteogenic medium + IL-1β (1 ng/ml) + PEI-pIL-1Ra nanoparticles. Diameter of each cylindrical reconstruction is 4 mm, while height is 3 mm.
    Figure Legend Snippet: Protective effect of PEI-pIL-1Ra nanoparticles on IL-1β-mediated inhibition of osteogenesis in BM-MSCs. Representative 3D microCT reconstructions depicting mineral deposition by BM-MSCs after 21 days in CHA scaffolds cultured under (A) basal medium, (B) osteogenic medium, (C) osteogenic medium + IL-1β (1 ng/ml), and (D) osteogenic medium + IL-1β (1 ng/ml) + PEI-pIL-1Ra nanoparticles. Diameter of each cylindrical reconstruction is 4 mm, while height is 3 mm.

    Techniques Used: Inhibition, Cell Culture

    Effect of transfection with PEI-pIL-1Ra on rat BM-MSCs cultured in presence of IL-1β. Cells were transfected with either PEI-pGFP or PEI-pIL-1Ra and cultured with osteogenic media (OM) + IL-1β (1 ng/ml). For comparison, BM-MSCs were also cultured with OM, OM + IL-1β (1 ng/ml), and OM + IL-1β (1 ng/ml) + IL-1Ra (10 ng/ml). The effect of transfection with PEI-pIL-1Ra was assessed in terms of (A) Alpl gene expression, (B) Ibsp gene expression, and (C) mineral content deposition. *, **, ***, and **** denotes p
    Figure Legend Snippet: Effect of transfection with PEI-pIL-1Ra on rat BM-MSCs cultured in presence of IL-1β. Cells were transfected with either PEI-pGFP or PEI-pIL-1Ra and cultured with osteogenic media (OM) + IL-1β (1 ng/ml). For comparison, BM-MSCs were also cultured with OM, OM + IL-1β (1 ng/ml), and OM + IL-1β (1 ng/ml) + IL-1Ra (10 ng/ml). The effect of transfection with PEI-pIL-1Ra was assessed in terms of (A) Alpl gene expression, (B) Ibsp gene expression, and (C) mineral content deposition. *, **, ***, and **** denotes p

    Techniques Used: Transfection, Cell Culture, Expressing

    Characterization of BM-MSCs transfection with PEI-pDNA nanoparticles. BM-MSCs were transfected with PEI-GLuc and compared to non-transfected (N/T) in terms of (A) metabolic activity as a surrogate marker of cell viability, and (B) in terms of luciferase expression. BM-MSCs were also transfected with pGFP nanoparticles to determine their (C) transfection efficiency over time, while (D) brightfield and fluorescence microscopy image overlay shows GFP + , successfully transfected cells. *, ***, and **** denotes p
    Figure Legend Snippet: Characterization of BM-MSCs transfection with PEI-pDNA nanoparticles. BM-MSCs were transfected with PEI-GLuc and compared to non-transfected (N/T) in terms of (A) metabolic activity as a surrogate marker of cell viability, and (B) in terms of luciferase expression. BM-MSCs were also transfected with pGFP nanoparticles to determine their (C) transfection efficiency over time, while (D) brightfield and fluorescence microscopy image overlay shows GFP + , successfully transfected cells. *, ***, and **** denotes p

    Techniques Used: Transfection, Activity Assay, Marker, Luciferase, Expressing, Fluorescence, Microscopy

    Assessment of BM-MSCs transfection in gene-activated CHA scaffolds. Cell viability of BM-MSCs in CHA scaffolds activated with PEI-pIL-1Ra nanoparticles after 3 days in culture is depicted in (A–C) : (A) Dead cells (ethidium homodimer 1-positive), (B) live cells (calcein-positive), (C) Overlay of live and dead cells. (D) Metabolic activity of cells in CHA scaffolds without nanoparticles (non-transfected) and CHA scaffolds activated with PEI-pIL-1Ra nanoparticles (PEI) after 3, 7, 10, and 14 days. (E) GFP + cells were observed in CHA scaffolds activated with PEI-pIL-1Ra nanoparticles 3 days post-transfection. (F) The transgene expression profile of luciferase in CHA scaffolds activated with PEI-pGLuc nanoparticles, normalized to luciferase expression 3 days post-transfection. *, **, and *** denotes p
    Figure Legend Snippet: Assessment of BM-MSCs transfection in gene-activated CHA scaffolds. Cell viability of BM-MSCs in CHA scaffolds activated with PEI-pIL-1Ra nanoparticles after 3 days in culture is depicted in (A–C) : (A) Dead cells (ethidium homodimer 1-positive), (B) live cells (calcein-positive), (C) Overlay of live and dead cells. (D) Metabolic activity of cells in CHA scaffolds without nanoparticles (non-transfected) and CHA scaffolds activated with PEI-pIL-1Ra nanoparticles (PEI) after 3, 7, 10, and 14 days. (E) GFP + cells were observed in CHA scaffolds activated with PEI-pIL-1Ra nanoparticles 3 days post-transfection. (F) The transgene expression profile of luciferase in CHA scaffolds activated with PEI-pGLuc nanoparticles, normalized to luciferase expression 3 days post-transfection. *, **, and *** denotes p

    Techniques Used: Transfection, Activity Assay, Expressing, Luciferase

    10) Product Images from "Characterization and evaluation of mesenchymal stem cells derived from human embryonic stem cells and bone marrow"

    Article Title: Characterization and evaluation of mesenchymal stem cells derived from human embryonic stem cells and bone marrow

    Journal: Cell and tissue research

    doi: 10.1007/s00441-014-1926-5

    Evaluation of chondrogenesis of ESC-MSCs and BM-MSCs. The mRNA expression of cartilage-related markers, collagen types 2 ( a ), and 9 ( b ), aggrecan ( c ), SOX9 ( d ), and collagen types 1 ( e ) and 10 ( f ) of ESC-MSC and BM-MSC pellets was analyzed by qPCR during 21 days of chondrogenic differentiation. * p
    Figure Legend Snippet: Evaluation of chondrogenesis of ESC-MSCs and BM-MSCs. The mRNA expression of cartilage-related markers, collagen types 2 ( a ), and 9 ( b ), aggrecan ( c ), SOX9 ( d ), and collagen types 1 ( e ) and 10 ( f ) of ESC-MSC and BM-MSC pellets was analyzed by qPCR during 21 days of chondrogenic differentiation. * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Analysis of growth and senescence of ESC-MSCs and BM-MSCs in expansion culture. a Long-term cell growth of H1-MSC, H9-MSC and BM-MSC cultures was indicated by cumulative population doublings calculated based on cell numbers measured during the culture period. MSC lines were derived from H1 ( diamond ), H9 ( filled circle ) and BM ( square, triangle , and cross ). Analysis of cellular senescence in long-term culture was performed on H1-MSCs, H9-MSCs and BM-MSCs at passages 2 ( b–f ), 7 ( g–k ), and 13 ( l–p ) by staining β-galactosidase activity of the cells. q Short-term proliferation ofH1-MSCs,H9-MSCs,WA1-MSCs and BM-MSC1 during a 9-day culture period was analyzed. Samples harvested on days 3, 6, and 9 were analyzed using PicoGreen. * p
    Figure Legend Snippet: Analysis of growth and senescence of ESC-MSCs and BM-MSCs in expansion culture. a Long-term cell growth of H1-MSC, H9-MSC and BM-MSC cultures was indicated by cumulative population doublings calculated based on cell numbers measured during the culture period. MSC lines were derived from H1 ( diamond ), H9 ( filled circle ) and BM ( square, triangle , and cross ). Analysis of cellular senescence in long-term culture was performed on H1-MSCs, H9-MSCs and BM-MSCs at passages 2 ( b–f ), 7 ( g–k ), and 13 ( l–p ) by staining β-galactosidase activity of the cells. q Short-term proliferation ofH1-MSCs,H9-MSCs,WA1-MSCs and BM-MSC1 during a 9-day culture period was analyzed. Samples harvested on days 3, 6, and 9 were analyzed using PicoGreen. * p

    Techniques Used: Derivative Assay, Staining, Activity Assay

    Evaluation of immunomodulation of ESC-MSCs and BM-MSCs on PBMCs. PBMCs were co-cultured with BM-MSCs or ESC-MSCs in different compartments of a transwell plate and stimulated with IL2. PBMCs were collected on days 3 and 6 and quantified by PicoGreen. * p
    Figure Legend Snippet: Evaluation of immunomodulation of ESC-MSCs and BM-MSCs on PBMCs. PBMCs were co-cultured with BM-MSCs or ESC-MSCs in different compartments of a transwell plate and stimulated with IL2. PBMCs were collected on days 3 and 6 and quantified by PicoGreen. * p

    Techniques Used: Cell Culture

    Evaluation of osteogenesis and adipogenesis of ESC-MSCs and BM-MSCs. Matrix mineralization during osteogenesis of BM-MSCs and ESC-MSCs was detected by Alizarin red S after 24 days of culture ( a–f ). The mRNA expression levels of bone-related markers alkaline phosphatase ( g ), CBFA1/RUNX2 ( h ) and osteocalcin ( i ) of ESC-MSCs and BM-MSCs during osteogenic differentiation were determined by qPCR. * p
    Figure Legend Snippet: Evaluation of osteogenesis and adipogenesis of ESC-MSCs and BM-MSCs. Matrix mineralization during osteogenesis of BM-MSCs and ESC-MSCs was detected by Alizarin red S after 24 days of culture ( a–f ). The mRNA expression levels of bone-related markers alkaline phosphatase ( g ), CBFA1/RUNX2 ( h ) and osteocalcin ( i ) of ESC-MSCs and BM-MSCs during osteogenic differentiation were determined by qPCR. * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Characterization of growth factor receptors and evaluation of chondrogenesis of ESC-MSC pellets. a Changes in the expression levels of ACVR2B, BMPR1B, and IGF1R during the first 6 days of ESC-MSC chondrogenesis were analyzed by flow cytometry. The mRNA expression of cartilage-related markers of ESC-MSC pellets induced by TGFβ1 and BMP7 (TB) for 21 days was compared to those of ESC-MSCs treated with TGFβ1 (T) ( b–d ). * p
    Figure Legend Snippet: Characterization of growth factor receptors and evaluation of chondrogenesis of ESC-MSC pellets. a Changes in the expression levels of ACVR2B, BMPR1B, and IGF1R during the first 6 days of ESC-MSC chondrogenesis were analyzed by flow cytometry. The mRNA expression of cartilage-related markers of ESC-MSC pellets induced by TGFβ1 and BMP7 (TB) for 21 days was compared to those of ESC-MSCs treated with TGFβ1 (T) ( b–d ). * p

    Techniques Used: Expressing, Flow Cytometry, Cytometry

    Characterization of morphology and immunophenotype of human ESC-MSCs and BM-MSCs in expansion culture. Morphology of MSCs was analyzed using bright field microscopy ( a–d ). e Flow cytometry of ESC-MSCs and BM-MSCs was performed with PE-conjugated antibodies to detect cell surface markers. The expression of isotype controls are shown as red histograms . f The expression of growth factor receptors of ESC-MSCs and BM-MSCs was analyzed by flow cytometry using PE-conjugated antibodies. Scale bars 400 µm
    Figure Legend Snippet: Characterization of morphology and immunophenotype of human ESC-MSCs and BM-MSCs in expansion culture. Morphology of MSCs was analyzed using bright field microscopy ( a–d ). e Flow cytometry of ESC-MSCs and BM-MSCs was performed with PE-conjugated antibodies to detect cell surface markers. The expression of isotype controls are shown as red histograms . f The expression of growth factor receptors of ESC-MSCs and BM-MSCs was analyzed by flow cytometry using PE-conjugated antibodies. Scale bars 400 µm

    Techniques Used: Microscopy, Flow Cytometry, Cytometry, Expressing

    11) Product Images from "Targeting ectodysplasin promotor by CRISPR/dCas9-effector effectively induces the reprogramming of human bone marrow-derived mesenchymal stem cells into sweat gland-like cells"

    Article Title: Targeting ectodysplasin promotor by CRISPR/dCas9-effector effectively induces the reprogramming of human bone marrow-derived mesenchymal stem cells into sweat gland-like cells

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-017-0758-0

    The effect of Dox-induced dCas9-E BM-MSCs on wound healing in vivo. a The mice were treated with Dox-induced dCas9-E BM-MSCs after full thickness scald injuries. b Sirius Red staining and Masson staining were used to detect the fibrosis, and the arrangement of collagen fibers was more regular than in the sham-treated group. Immunostaining was performed for c Ki67 (green) and d CD31 (red) after treatment with Dox-induced dCas9-E BM-MSCs. Scale bars = 20 μm. DAPI 4’6-diamidino-2-phenylindole, sgRNA single-guide RNA
    Figure Legend Snippet: The effect of Dox-induced dCas9-E BM-MSCs on wound healing in vivo. a The mice were treated with Dox-induced dCas9-E BM-MSCs after full thickness scald injuries. b Sirius Red staining and Masson staining were used to detect the fibrosis, and the arrangement of collagen fibers was more regular than in the sham-treated group. Immunostaining was performed for c Ki67 (green) and d CD31 (red) after treatment with Dox-induced dCas9-E BM-MSCs. Scale bars = 20 μm. DAPI 4’6-diamidino-2-phenylindole, sgRNA single-guide RNA

    Techniques Used: In Vivo, Mouse Assay, Staining, Immunostaining

    Involvement of NF-κB pathways in dCas9-E-induced EDA expression. The cells transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h exhibited a significance difference in NF-κB downstream Sonic Hedgehog (Shh) and cyclin D1 mRNA ( a ) and protein ( b ) expression. Immunostaining was also performed to detect the activity of the NF-κB pathway. dCas9-E BM-MSCs induced more NF-κB translocated into the nuclei and then activated the expression of Shh and cyclin D1 ( c ). All data are normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and calibrated based on the BM-MSC group. *** p
    Figure Legend Snippet: Involvement of NF-κB pathways in dCas9-E-induced EDA expression. The cells transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h exhibited a significance difference in NF-κB downstream Sonic Hedgehog (Shh) and cyclin D1 mRNA ( a ) and protein ( b ) expression. Immunostaining was also performed to detect the activity of the NF-κB pathway. dCas9-E BM-MSCs induced more NF-κB translocated into the nuclei and then activated the expression of Shh and cyclin D1 ( c ). All data are normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and calibrated based on the BM-MSC group. *** p

    Techniques Used: Expressing, Transfection, Incubation, Immunostaining, Activity Assay

    The effect of dCas9-E BM-MSCs on the regeneration of sweat glands in vivo. a The saline-treated paw (Sham) showed a negative result for the perspiration test, whereas paws implanted with Dox-induced dCas9-E BM-MSCs showed a positive result (sgRNA2). b Hematoxylin and eosin staining was used to detect the duct structure of sweat gland tissue after injection with Dox-induced dCas9-E BM-MSCs. c The expression of carcinoembryonic antigen (CEA; red), cytokeratin 19 (CK19; green) and nucleus (DAPI) in BM-MSCs after implantation for 20 days. d The quantification of each paw (0.5 cm × 0.5 cm, treated with collagenase I for 1 h at 37 °C) was also conducted after treatment with Dox-induced dCas9-E BM-MSCs. ** p
    Figure Legend Snippet: The effect of dCas9-E BM-MSCs on the regeneration of sweat glands in vivo. a The saline-treated paw (Sham) showed a negative result for the perspiration test, whereas paws implanted with Dox-induced dCas9-E BM-MSCs showed a positive result (sgRNA2). b Hematoxylin and eosin staining was used to detect the duct structure of sweat gland tissue after injection with Dox-induced dCas9-E BM-MSCs. c The expression of carcinoembryonic antigen (CEA; red), cytokeratin 19 (CK19; green) and nucleus (DAPI) in BM-MSCs after implantation for 20 days. d The quantification of each paw (0.5 cm × 0.5 cm, treated with collagenase I for 1 h at 37 °C) was also conducted after treatment with Dox-induced dCas9-E BM-MSCs. ** p

    Techniques Used: In Vivo, Staining, Injection, Expressing

    dCas9-E expression in BM-MSCs. a Bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with pLKO.1-puro-U6 and dCas9-E, and stained with PE-labeled anti-HA and DAPI 72 h post-transfection. b The expression of designed dCas9-E nucleases. Scale bar = 50 μm. DAPI 4’6-diamidino-2-phenylindole, GAPDH glyceraldehyde-3-phosphate dehydrogenase, HA hemagglutinin, sgRNA single-guide RNA
    Figure Legend Snippet: dCas9-E expression in BM-MSCs. a Bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with pLKO.1-puro-U6 and dCas9-E, and stained with PE-labeled anti-HA and DAPI 72 h post-transfection. b The expression of designed dCas9-E nucleases. Scale bar = 50 μm. DAPI 4’6-diamidino-2-phenylindole, GAPDH glyceraldehyde-3-phosphate dehydrogenase, HA hemagglutinin, sgRNA single-guide RNA

    Techniques Used: Expressing, Derivative Assay, Transfection, Staining, Labeling

    Characteristics of dCas9-E BM-MSCs after Dox induction. a The sweat gland markers, carcinoembryonic antigen (CEA) and cytokeratin (CK)19, were detected by qRT-PCR after bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h. b Proteins were collected from BM-MSCs transfected with dCas9-E. The sweat glands biomarkers CEA, CK7, CK14, and CK19 were detected by Western blotting using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for calibration of sample loading. c Immunofluorescence staining was used to detect CEA (red) and CK19 (green) in BM-MSCs (scale bar = 50 μm). d Villi ultrastructure changes after dCas9-E BM-MSCs were treated with Dox (scale bar = 1 μm). ** p
    Figure Legend Snippet: Characteristics of dCas9-E BM-MSCs after Dox induction. a The sweat gland markers, carcinoembryonic antigen (CEA) and cytokeratin (CK)19, were detected by qRT-PCR after bone marrow-derived mesenchymal stem cells (BM-MSCs) were transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h. b Proteins were collected from BM-MSCs transfected with dCas9-E. The sweat glands biomarkers CEA, CK7, CK14, and CK19 were detected by Western blotting using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for calibration of sample loading. c Immunofluorescence staining was used to detect CEA (red) and CK19 (green) in BM-MSCs (scale bar = 50 μm). d Villi ultrastructure changes after dCas9-E BM-MSCs were treated with Dox (scale bar = 1 μm). ** p

    Techniques Used: Quantitative RT-PCR, Derivative Assay, Transfection, Incubation, Western Blot, Immunofluorescence, Staining

    dCas9-E mediated activation of EDA in BM-MSCs. The cells were harvested and total mRNA and protein were extracted. Ectodysplasin (EDA) gene and protein expression were identified by qPCR ( a ) and Western blotting ( b ) after doxycycline (Dox) induction for 48 h. c Fluorescence microscopy detection of EDA expression activated by dCas9-E. After supplementing with 2 μg/ml Dox for 48 h, enhanced expression of EDA (red) was detected in dCas9-E transfected bone marrow-derived mesenchymal stem cells (BM-MSCs). The data are expressed as the mean ± SD. *** p
    Figure Legend Snippet: dCas9-E mediated activation of EDA in BM-MSCs. The cells were harvested and total mRNA and protein were extracted. Ectodysplasin (EDA) gene and protein expression were identified by qPCR ( a ) and Western blotting ( b ) after doxycycline (Dox) induction for 48 h. c Fluorescence microscopy detection of EDA expression activated by dCas9-E. After supplementing with 2 μg/ml Dox for 48 h, enhanced expression of EDA (red) was detected in dCas9-E transfected bone marrow-derived mesenchymal stem cells (BM-MSCs). The data are expressed as the mean ± SD. *** p

    Techniques Used: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Fluorescence, Microscopy, Transfection, Derivative Assay

    12) Product Images from "MSX2 Initiates and Accelerates Mesenchymal Stem/Stromal Cell Specification of hPSCs by Regulating TWIST1 and PRAME"

    Article Title: MSX2 Initiates and Accelerates Mesenchymal Stem/Stromal Cell Specification of hPSCs by Regulating TWIST1 and PRAME

    Journal: Stem Cell Reports

    doi: 10.1016/j.stemcr.2018.06.019

    MC-MSCs Resemble BM-MSCs and Are Functional (A) Hierarchical clustering analysis of hPSCs (H1 hESCs), M-MSCs, MC-MSCs, BM-MSCs. (B) Heatmap illustrating expression of mesenchymal development/differentiation and pluripotency-associated genes for hPSCs (H1 hESCs), M-MSCs, MC-MSCs, BM-MSCs. (C) GSEA comparing MC-MSCs and hPSCs (H1 hESCs). The NES and p values are shown. (D) The sorted CD3 + T lymphocytes were stimulated with plate-bound anti-CD3 antibody and anti-CD28 antibody or with Molecular Probes sulfate latex for 72 hr. Then, the lymphocytes were stained with anti-CD4 or anti-CD8 antibodies for CFSE dilution analysis. One of three independent experiments is shown. Ctr, control. (E) Mice were given untreated drinking water (control) or 2% DSS in drinking water (DSS) for 6 days ( Wang et al., 2016 ). Then, all mice were given untreated drinking water for the next 7 days. On days 2 and 3, mice treated with DSS were injected intraperitoneally (i.p.) with PBS, MC-MSCs or BM-MSCs. The control group mice were injected i.p. with PBS. The change in body weight of mice was measured. Data are analyzed by multiple t test and shown as mean ± SEM (N = 3). ∗ p
    Figure Legend Snippet: MC-MSCs Resemble BM-MSCs and Are Functional (A) Hierarchical clustering analysis of hPSCs (H1 hESCs), M-MSCs, MC-MSCs, BM-MSCs. (B) Heatmap illustrating expression of mesenchymal development/differentiation and pluripotency-associated genes for hPSCs (H1 hESCs), M-MSCs, MC-MSCs, BM-MSCs. (C) GSEA comparing MC-MSCs and hPSCs (H1 hESCs). The NES and p values are shown. (D) The sorted CD3 + T lymphocytes were stimulated with plate-bound anti-CD3 antibody and anti-CD28 antibody or with Molecular Probes sulfate latex for 72 hr. Then, the lymphocytes were stained with anti-CD4 or anti-CD8 antibodies for CFSE dilution analysis. One of three independent experiments is shown. Ctr, control. (E) Mice were given untreated drinking water (control) or 2% DSS in drinking water (DSS) for 6 days ( Wang et al., 2016 ). Then, all mice were given untreated drinking water for the next 7 days. On days 2 and 3, mice treated with DSS were injected intraperitoneally (i.p.) with PBS, MC-MSCs or BM-MSCs. The control group mice were injected i.p. with PBS. The change in body weight of mice was measured. Data are analyzed by multiple t test and shown as mean ± SEM (N = 3). ∗ p

    Techniques Used: Functional Assay, Expressing, Staining, Mouse Assay, Injection

    13) Product Images from "Comparison of Immunosuppressive and Angiogenic Properties of Human Amnion-Derived Mesenchymal Stem Cells between 2D and 3D Culture Systems"

    Article Title: Comparison of Immunosuppressive and Angiogenic Properties of Human Amnion-Derived Mesenchymal Stem Cells between 2D and 3D Culture Systems

    Journal: Stem Cells International

    doi: 10.1155/2019/7486279

    Inhibition of activated PBMCs. (a) Representative images of unstimulated (No ACT) and CD3/CD28-stimulated (ACT) PBMCs grown in each conditioned medium. (b) FACS analysis of PBMC after 4 days of culture in each conditioned medium. DMEM without CD3/CD28 (No ACT). DMEM with CD3/CD28 (ACT). DMEM conditioned by HDFa (HDFa CM). DMEM conditioned by BM-MSCs (BM-MSC CM). Exosome-depleted DMEM conditioned by hAMSCs grown as monolayer (2D CM-exo). DMEM conditioned by hAMSCs grown as monolayer (2D CM). Exosome-depleted DMEM conditioned by hAMSCs grown as spheroids (3D CM-exo). DMEM conditioned by hAMSCs grown as spheroids (3D CM). 5 μ g/ml exosomes secreted by hAMSCs grown as monolayer (2D EXO 5 μ g/ml). 5 μ g/ml exosomes secreted by hAMSCs grown as spheroids (3D EXO 5 μ g/ml). Data are means ± SD of triplicate in three independent experiments. ∗ p
    Figure Legend Snippet: Inhibition of activated PBMCs. (a) Representative images of unstimulated (No ACT) and CD3/CD28-stimulated (ACT) PBMCs grown in each conditioned medium. (b) FACS analysis of PBMC after 4 days of culture in each conditioned medium. DMEM without CD3/CD28 (No ACT). DMEM with CD3/CD28 (ACT). DMEM conditioned by HDFa (HDFa CM). DMEM conditioned by BM-MSCs (BM-MSC CM). Exosome-depleted DMEM conditioned by hAMSCs grown as monolayer (2D CM-exo). DMEM conditioned by hAMSCs grown as monolayer (2D CM). Exosome-depleted DMEM conditioned by hAMSCs grown as spheroids (3D CM-exo). DMEM conditioned by hAMSCs grown as spheroids (3D CM). 5 μ g/ml exosomes secreted by hAMSCs grown as monolayer (2D EXO 5 μ g/ml). 5 μ g/ml exosomes secreted by hAMSCs grown as spheroids (3D EXO 5 μ g/ml). Data are means ± SD of triplicate in three independent experiments. ∗ p

    Techniques Used: Inhibition, Activated Clotting Time Assay, FACS

    Expression analysis of angiogenic and immunosuppressive factors. Both gene (a, b) expression and protein (c, d) expression were assayed after 3 days of cultures in cells and CM, respectively. (a) Gene expression of angiogenic factor. (b) Gene expression of immunosuppressive factor. (c) Protein expression of angiogenic factor. (d) Protein expression of immunosuppressive factor. (e) Hierarchical clustering of gene expression profile. Transcript levels were normalized to those of GAPDH and expressed as fold change vs. gene expression values of HDFa. Bone marrow mesenchymal stem cells (BM-MSCs). Amnion mesenchymal stem cells grown in two-dimensional cultures (2D hAMSCs). Amnion mesenchymal stem cells grown in three-dimensional cultures (3D hAMSCs). DMEM conditioned by BM-MSCs (BM-MSC CM). Exosome-depleted DMEM conditioned by hAMSCs grown as monolayer (2D CM-exo). DMEM conditioned by hAMSCs grown as monolayer (2D CM). Exosome-depleted DMEM conditioned by hAMSCs grown as spheroids (3D CM-exo). DMEM conditioned by hAMSCs grown as spheroids (3D CM). Data are means ± SD of triplicate in three independent experiments. ∗ p
    Figure Legend Snippet: Expression analysis of angiogenic and immunosuppressive factors. Both gene (a, b) expression and protein (c, d) expression were assayed after 3 days of cultures in cells and CM, respectively. (a) Gene expression of angiogenic factor. (b) Gene expression of immunosuppressive factor. (c) Protein expression of angiogenic factor. (d) Protein expression of immunosuppressive factor. (e) Hierarchical clustering of gene expression profile. Transcript levels were normalized to those of GAPDH and expressed as fold change vs. gene expression values of HDFa. Bone marrow mesenchymal stem cells (BM-MSCs). Amnion mesenchymal stem cells grown in two-dimensional cultures (2D hAMSCs). Amnion mesenchymal stem cells grown in three-dimensional cultures (3D hAMSCs). DMEM conditioned by BM-MSCs (BM-MSC CM). Exosome-depleted DMEM conditioned by hAMSCs grown as monolayer (2D CM-exo). DMEM conditioned by hAMSCs grown as monolayer (2D CM). Exosome-depleted DMEM conditioned by hAMSCs grown as spheroids (3D CM-exo). DMEM conditioned by hAMSCs grown as spheroids (3D CM). Data are means ± SD of triplicate in three independent experiments. ∗ p

    Techniques Used: Expressing

    HUVEC migration assay and capillary-like formation assay. (a) Real-time migration monitoring of HUVECs with the xCELLigence system. (b) Slopes of migration curves. (c) Representative images of HUVECs on Matrigel, in contact with each conditioned medium. (d-f) Graphs represent a quantitative analysis of capillary length (d), branching point number (e), and node number (F). DMEM serum-free medium (DMEM). DMEM conditioned by HDFa (HDFa CM). DMEM conditioned by BM-MSCs (BM-MSC CM). Exosome-depleted DMEM conditioned by hAMSCs grown as monolayer (2D CM-exo). DMEM conditioned by hAMSCs grown as monolayer (2D CM). Exosome-depleted DMEM conditioned by hAMSCs grown as spheroids (3D CM-exo). DMEM conditioned by hAMSCs grown as spheroids (3D CM). 5 μ g/ml exosomes secreted by hAMSCs grown as monolayer (2D EXO 5 μ g/ml). 5 μ g/ml exosomes secreted by hAMSCs grown as spheroids (3D EXO 5 μ g/ml). Data are means ± SD of quadruplicate in three independent experiments. ∗ p
    Figure Legend Snippet: HUVEC migration assay and capillary-like formation assay. (a) Real-time migration monitoring of HUVECs with the xCELLigence system. (b) Slopes of migration curves. (c) Representative images of HUVECs on Matrigel, in contact with each conditioned medium. (d-f) Graphs represent a quantitative analysis of capillary length (d), branching point number (e), and node number (F). DMEM serum-free medium (DMEM). DMEM conditioned by HDFa (HDFa CM). DMEM conditioned by BM-MSCs (BM-MSC CM). Exosome-depleted DMEM conditioned by hAMSCs grown as monolayer (2D CM-exo). DMEM conditioned by hAMSCs grown as monolayer (2D CM). Exosome-depleted DMEM conditioned by hAMSCs grown as spheroids (3D CM-exo). DMEM conditioned by hAMSCs grown as spheroids (3D CM). 5 μ g/ml exosomes secreted by hAMSCs grown as monolayer (2D EXO 5 μ g/ml). 5 μ g/ml exosomes secreted by hAMSCs grown as spheroids (3D EXO 5 μ g/ml). Data are means ± SD of quadruplicate in three independent experiments. ∗ p

    Techniques Used: Migration, Tube Formation Assay

    14) Product Images from "N-cadherin mediates the migration of bone marrow-derived mesenchymal stem cells toward breast tumor cells"

    Article Title: N-cadherin mediates the migration of bone marrow-derived mesenchymal stem cells toward breast tumor cells

    Journal: Theranostics

    doi: 10.7150/thno.59703

    N-cadherin mediates the migration of bone marrow-derived mesenchymal stem cells (BM-MSCs) toward MCF-7 or MDA-MB-231 conditioned medium. (A) Verification of N-cadherin knockdown in BM-MSCs transfected with siRNA that were used in the migration assay with the MCF7 conditioned medium (MCF7 CM). (B-C) Migration of BM-MSCs transfected with each siRNA in response to MCF7 CM. BM-MSCs transfected with control siRNA (siCON) or N-cadherin siRNA (siN-cad) were treated with the control conditioned medium (CON CM) or MCF7 CM for 12 h. (D) Verification of N-cadherin knockdown in BM-MSCs transfected with siRNA that were used in the migration assay with the MDA-MB-231 conditioned medium (MDA CM). (E-F) Migration of BM-MSCs transfected with each siRNA in response to MDA CM. BM-MSCs transfected with each siRNA were treated with CON CM or MDA CM for 12 h. White arrows indicate DAPI-stained nuclei of migrated cells on the lower membrane surface. The red lines indicate the mean values ( n = 2 samples for each group). Scale bars indicate 100 μm.
    Figure Legend Snippet: N-cadherin mediates the migration of bone marrow-derived mesenchymal stem cells (BM-MSCs) toward MCF-7 or MDA-MB-231 conditioned medium. (A) Verification of N-cadherin knockdown in BM-MSCs transfected with siRNA that were used in the migration assay with the MCF7 conditioned medium (MCF7 CM). (B-C) Migration of BM-MSCs transfected with each siRNA in response to MCF7 CM. BM-MSCs transfected with control siRNA (siCON) or N-cadherin siRNA (siN-cad) were treated with the control conditioned medium (CON CM) or MCF7 CM for 12 h. (D) Verification of N-cadherin knockdown in BM-MSCs transfected with siRNA that were used in the migration assay with the MDA-MB-231 conditioned medium (MDA CM). (E-F) Migration of BM-MSCs transfected with each siRNA in response to MDA CM. BM-MSCs transfected with each siRNA were treated with CON CM or MDA CM for 12 h. White arrows indicate DAPI-stained nuclei of migrated cells on the lower membrane surface. The red lines indicate the mean values ( n = 2 samples for each group). Scale bars indicate 100 μm.

    Techniques Used: Migration, Derivative Assay, Multiple Displacement Amplification, Transfection, Staining

    N-cadherin is necessary for TGF-β-mediated migration of bone marrow-derived mesenchymal stem cells (BM-MSCs). (A-B) Western blot analysis of N-cadherin and ɑ-tubulin and qRT-PCR analysis of N-cadherin. BM-MSCs transfected with control siRNA (siCON) or N‐cadherin siRNA (siN‐cad) were treated with TGF‐β (1 ng/mL) for 24 h. Results are presented as mean ± SD. (C-D) Migration of BM-MSCs transfected with each siRNA in response to TGF‐β (10 ng/mL). (E-F) Migration of BM-MSCs pretreated with EGTA for 30 min in response to TGF‐β (10 ng/mL). White arrows indicate DAPI-stained nuclei of migrated BM-MSCs on the lower membrane surface. The red lines indicate the mean values ( n = 2 samples for each group). Scale bars indicate 100 μm.
    Figure Legend Snippet: N-cadherin is necessary for TGF-β-mediated migration of bone marrow-derived mesenchymal stem cells (BM-MSCs). (A-B) Western blot analysis of N-cadherin and ɑ-tubulin and qRT-PCR analysis of N-cadherin. BM-MSCs transfected with control siRNA (siCON) or N‐cadherin siRNA (siN‐cad) were treated with TGF‐β (1 ng/mL) for 24 h. Results are presented as mean ± SD. (C-D) Migration of BM-MSCs transfected with each siRNA in response to TGF‐β (10 ng/mL). (E-F) Migration of BM-MSCs pretreated with EGTA for 30 min in response to TGF‐β (10 ng/mL). White arrows indicate DAPI-stained nuclei of migrated BM-MSCs on the lower membrane surface. The red lines indicate the mean values ( n = 2 samples for each group). Scale bars indicate 100 μm.

    Techniques Used: Migration, Derivative Assay, Western Blot, Quantitative RT-PCR, Transfection, Staining

    TGF-β increases N-cadherin expression in bone marrow-derived mesenchymal stem cells (BM-MSCs) in a Smad4-dependent manner. (A) Western blot analysis of phosphorylated Smad2/3 (p-Smad2/3), Smad2, phosphorylated Akt (p-Akt), Akt, phosphorylated ERKs (p-ERKs), ERKs, phosphorylated p38 (p-p38), p38, N-cadherin, and ɑ-tubulin. BM‐MSCs were treated with TGF‐β (1 ng/mL) for the indicated times. (B-C) Western blot analysis of N-cadherin, Smad4, and ɑ-tubulin. BM-MSCs transfected with control siRNA (siCON) or Smad4 siRNA (siSMAD4) were treated with TGF‐β (1 ng/mL) for 24 h. Densitometric analysis of western blot results in (C) . Results are presented as mean ± SD (five independent experiments). (D-E) qRT-PCR analysis of Smad4 and N-cadherin. BM-MSCs transfected with each siRNA were treated with TGF‐β (1 ng/mL) for 24 h. (F-H) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs were pretreated with LY294002 (2 μM), PD98059 (10 μM), or SB203580 (10 μM) for 30 min prior to treatment with TGF-β (1 ng/mL) for 24 h. (I-J) Migration of BM-MSCs transfected with each siRNA in response to TGF‐β (10 ng/mL). White arrows indicate DAPI-stained nuclei of migrated BM-MSCs on the lower membrane surface. Quantification of the results of migration assay in (J) . The red lines indicate the mean values ( n = 2 samples for each group). Scale bar indicates 100 μm.
    Figure Legend Snippet: TGF-β increases N-cadherin expression in bone marrow-derived mesenchymal stem cells (BM-MSCs) in a Smad4-dependent manner. (A) Western blot analysis of phosphorylated Smad2/3 (p-Smad2/3), Smad2, phosphorylated Akt (p-Akt), Akt, phosphorylated ERKs (p-ERKs), ERKs, phosphorylated p38 (p-p38), p38, N-cadherin, and ɑ-tubulin. BM‐MSCs were treated with TGF‐β (1 ng/mL) for the indicated times. (B-C) Western blot analysis of N-cadherin, Smad4, and ɑ-tubulin. BM-MSCs transfected with control siRNA (siCON) or Smad4 siRNA (siSMAD4) were treated with TGF‐β (1 ng/mL) for 24 h. Densitometric analysis of western blot results in (C) . Results are presented as mean ± SD (five independent experiments). (D-E) qRT-PCR analysis of Smad4 and N-cadherin. BM-MSCs transfected with each siRNA were treated with TGF‐β (1 ng/mL) for 24 h. (F-H) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs were pretreated with LY294002 (2 μM), PD98059 (10 μM), or SB203580 (10 μM) for 30 min prior to treatment with TGF-β (1 ng/mL) for 24 h. (I-J) Migration of BM-MSCs transfected with each siRNA in response to TGF‐β (10 ng/mL). White arrows indicate DAPI-stained nuclei of migrated BM-MSCs on the lower membrane surface. Quantification of the results of migration assay in (J) . The red lines indicate the mean values ( n = 2 samples for each group). Scale bar indicates 100 μm.

    Techniques Used: Expressing, Derivative Assay, Western Blot, Transfection, Quantitative RT-PCR, Migration, Staining

    Breast tumor cell-conditioned medium increases the expression of N-cadherin in bone marrow-derived mesenchymal stem cells (BM-MSCs) in a TGF-β-dependent manner. (A) The expression of TGF-β1 ligand in breast cancer compared with that in the normal tissue. The RNA-seq database of human breast cancers from The Cancer Genome Atlas (TCGA) was analyzed. The red lines indicate the mean values. (B) qRT-PCR analysis of TGFB1, a gene of TGF-β1 ligand in MCF7 and MDA-MB-231 cells. (C-D) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs were treated with the control conditioned medium (CON CM) or MCF7 conditioned medium (MCF7 CM) for 24 h. Densitometric analysis of western blot results in (three independent experiments) in (D) . (E-F) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs were treated with CON CM or the MDA-MB-231 conditioned medium (MDA CM) for 24 h. Densitometric analysis of western blot results (three independent experiments) in (F) . (G) qRT-PCR analysis of N-cadherin. BM-MSCs were treated with SB505124 (500 nM) for 30 min prior to treatment with CON CM or MDA CM for 24 h. (H) qRT-PCR analysis of TGFBR1, a gene of TGF-β type 1 receptor in BM-MSCs to verify the knockdown of TGFBR1. (I) Western blot analysis of phosphorylated Smad2/3 (p-Smad2/3) and Smad2. BM-MSCs transfected with control siRNA (siCON) or TGF-β type 1 receptor siRNA (siTGFBR1) were treated with TGF‐β (1 ng/mL) for 30 min. (J-K) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs transfected with each siRNA were treated with CON CM or MDA CM for 24 h. Densitometric analysis of western blot results (three independent experiments) in (K) . (L) qRT-PCR analysis of N-cadherin. BM-MSCs transfected with each siRNA were treated with CON CM or MDA CM for 24 h. Results are presented as the mean ± SD.
    Figure Legend Snippet: Breast tumor cell-conditioned medium increases the expression of N-cadherin in bone marrow-derived mesenchymal stem cells (BM-MSCs) in a TGF-β-dependent manner. (A) The expression of TGF-β1 ligand in breast cancer compared with that in the normal tissue. The RNA-seq database of human breast cancers from The Cancer Genome Atlas (TCGA) was analyzed. The red lines indicate the mean values. (B) qRT-PCR analysis of TGFB1, a gene of TGF-β1 ligand in MCF7 and MDA-MB-231 cells. (C-D) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs were treated with the control conditioned medium (CON CM) or MCF7 conditioned medium (MCF7 CM) for 24 h. Densitometric analysis of western blot results in (three independent experiments) in (D) . (E-F) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs were treated with CON CM or the MDA-MB-231 conditioned medium (MDA CM) for 24 h. Densitometric analysis of western blot results (three independent experiments) in (F) . (G) qRT-PCR analysis of N-cadherin. BM-MSCs were treated with SB505124 (500 nM) for 30 min prior to treatment with CON CM or MDA CM for 24 h. (H) qRT-PCR analysis of TGFBR1, a gene of TGF-β type 1 receptor in BM-MSCs to verify the knockdown of TGFBR1. (I) Western blot analysis of phosphorylated Smad2/3 (p-Smad2/3) and Smad2. BM-MSCs transfected with control siRNA (siCON) or TGF-β type 1 receptor siRNA (siTGFBR1) were treated with TGF‐β (1 ng/mL) for 30 min. (J-K) Western blot analysis of N-cadherin and ɑ-tubulin. BM-MSCs transfected with each siRNA were treated with CON CM or MDA CM for 24 h. Densitometric analysis of western blot results (three independent experiments) in (K) . (L) qRT-PCR analysis of N-cadherin. BM-MSCs transfected with each siRNA were treated with CON CM or MDA CM for 24 h. Results are presented as the mean ± SD.

    Techniques Used: Expressing, Derivative Assay, RNA Sequencing Assay, Quantitative RT-PCR, Multiple Displacement Amplification, Western Blot, Transfection

    N-cadherin-mediated cell-cell adhesion is required for the migration of bone marrow-derived mesenchymal stem cells (BM-MSCs) toward MDA-MB-231 conditioned medium. (A) Schematic representation of three-dimensional (3D) migration assay. (B-D) Three-dimensional migration of BM-MSCs toward the control conditioned medium (CON CM) or MDA-MB-231 conditioned medium (MDA CM). White dashed boxes in B' and C' are magnified in B'' and C''. Yellow broken lines indicate the margin of collagen gel and black broken lines represent the leading edge of migrating cells. Scale bars indicate 600 μm. Quantification of the results in (D) . The red lines indicate the mean values ( n = 3 samples for each group). (E) Three-dimensional migration of BM-MSCs transfected with control siRNA (siCON) or N-cadherin siRNA (siN-cad). The cells were treated with CON CM or MDA CM for 24 h. The red lines indicate the mean values ( n = 3 samples for each group). (F) Immunocytochemistry of the expression of N-cadherin and β-catenin in BM-MSCs undergoing 3D migration toward MDA CM. Actin was stained with phalloidin in red and nuclei were stained with DAPI in blue. White arrows indicate co-localization of N‐cadherin and β‐catenin at cell-cell adhesion borders of migrating BM-MSCs. Scale bar indicates 100 μm. (G-I) Three-dimensional migration using a mixture of BM-MSCs transfected with siCON (stained with calcein AM in green) or siN-cad. White dashed boxes in G' are magnified in G'' and yellow broken line indicates the margin of collagen gel. Green dotted lines appear around calcein AM stained BM-MSCs transfected with siCON and red dotted lines appear around BM-MSCs transfected with siN-cad. Scale bar indicates 100 μm. (H) Percentages of BM-MSCs transfected with siCON or BM-MSCs transfected with siN-cad among the total number of migrating BM-MSCs from each mixture of cell-collagen gel ( n = 3 ) . (I) The percentage of migrating cells maintaining cell-cell adhesion or migrating cells without cell-cell adhesion among the total number of migrating BM-MSCs transfected with siCON (N-cad+ cells) or the percentage of migrating cells maintaining cell-cell adhesion or migrating cells without cell-cell adhesion among the total number of migrating BM-MSCs transfected with siN-cads (N-cad- cells) (gray bars: the percentages of migrating cells with cell-cell adhesion; white bars: the percentages of migrating cell without cell-cell adhesion). Results are presented as the mean ± SD ( n = 3 mixtures of cell-collagen gel).
    Figure Legend Snippet: N-cadherin-mediated cell-cell adhesion is required for the migration of bone marrow-derived mesenchymal stem cells (BM-MSCs) toward MDA-MB-231 conditioned medium. (A) Schematic representation of three-dimensional (3D) migration assay. (B-D) Three-dimensional migration of BM-MSCs toward the control conditioned medium (CON CM) or MDA-MB-231 conditioned medium (MDA CM). White dashed boxes in B' and C' are magnified in B'' and C''. Yellow broken lines indicate the margin of collagen gel and black broken lines represent the leading edge of migrating cells. Scale bars indicate 600 μm. Quantification of the results in (D) . The red lines indicate the mean values ( n = 3 samples for each group). (E) Three-dimensional migration of BM-MSCs transfected with control siRNA (siCON) or N-cadherin siRNA (siN-cad). The cells were treated with CON CM or MDA CM for 24 h. The red lines indicate the mean values ( n = 3 samples for each group). (F) Immunocytochemistry of the expression of N-cadherin and β-catenin in BM-MSCs undergoing 3D migration toward MDA CM. Actin was stained with phalloidin in red and nuclei were stained with DAPI in blue. White arrows indicate co-localization of N‐cadherin and β‐catenin at cell-cell adhesion borders of migrating BM-MSCs. Scale bar indicates 100 μm. (G-I) Three-dimensional migration using a mixture of BM-MSCs transfected with siCON (stained with calcein AM in green) or siN-cad. White dashed boxes in G' are magnified in G'' and yellow broken line indicates the margin of collagen gel. Green dotted lines appear around calcein AM stained BM-MSCs transfected with siCON and red dotted lines appear around BM-MSCs transfected with siN-cad. Scale bar indicates 100 μm. (H) Percentages of BM-MSCs transfected with siCON or BM-MSCs transfected with siN-cad among the total number of migrating BM-MSCs from each mixture of cell-collagen gel ( n = 3 ) . (I) The percentage of migrating cells maintaining cell-cell adhesion or migrating cells without cell-cell adhesion among the total number of migrating BM-MSCs transfected with siCON (N-cad+ cells) or the percentage of migrating cells maintaining cell-cell adhesion or migrating cells without cell-cell adhesion among the total number of migrating BM-MSCs transfected with siN-cads (N-cad- cells) (gray bars: the percentages of migrating cells with cell-cell adhesion; white bars: the percentages of migrating cell without cell-cell adhesion). Results are presented as the mean ± SD ( n = 3 mixtures of cell-collagen gel).

    Techniques Used: Migration, Derivative Assay, Multiple Displacement Amplification, Transfection, Immunocytochemistry, Expressing, Staining

    15) Product Images from "TNF?-exposed Bone Marrow-derived Mesenchymal Stem Cells Promote Locomotion of MDA-MB-231 Breast Cancer Cells through Transcriptional Activation of CXCR3 Ligand Chemokines *"

    Article Title: TNF?-exposed Bone Marrow-derived Mesenchymal Stem Cells Promote Locomotion of MDA-MB-231 Breast Cancer Cells through Transcriptional Activation of CXCR3 Ligand Chemokines *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.128124

    Relative expression of various cytokines and chemokines in response to TNFα stimulation in BM-MSCs. BM-MSCs were either left untreated or treated with 10 ng/ml TNFα for 24 h. Relative -fold changes of mRNA levels between control and TNFα-treated
    Figure Legend Snippet: Relative expression of various cytokines and chemokines in response to TNFα stimulation in BM-MSCs. BM-MSCs were either left untreated or treated with 10 ng/ml TNFα for 24 h. Relative -fold changes of mRNA levels between control and TNFα-treated

    Techniques Used: Expressing

    NF-κB binding site is essential for TNFα-induced promoter activation of CXCR3 ligand chemokines. BM-MSCs were transiently transfected with 0.2 μg of serial deletion or NF-κB site mutant ( mtNF κ B ) constructs of CXCL9
    Figure Legend Snippet: NF-κB binding site is essential for TNFα-induced promoter activation of CXCR3 ligand chemokines. BM-MSCs were transiently transfected with 0.2 μg of serial deletion or NF-κB site mutant ( mtNF κ B ) constructs of CXCL9

    Techniques Used: Binding Assay, Activation Assay, Transfection, Mutagenesis, Construct

    Association of NF-κB to CXCR3 ligand chemokine promoters. BM-MSCs were either untreated or treated with 10 ng/ml TNFα for 4 h, and nuclear extracts were prepared. EMSA was done with 32 P-labeled oligonucleotide probes corresponding to positions
    Figure Legend Snippet: Association of NF-κB to CXCR3 ligand chemokine promoters. BM-MSCs were either untreated or treated with 10 ng/ml TNFα for 4 h, and nuclear extracts were prepared. EMSA was done with 32 P-labeled oligonucleotide probes corresponding to positions

    Techniques Used: Labeling

    Up-regulation of CXCR3 ligand chemokines by TNFα in BM-MSCs. A , BM-MSCs were stimulated with TNFα (10 ng/ml) for various lengths of time (6–24 h). Total RNAs were prepared, and Northern blotting was performed. GAPDH expression
    Figure Legend Snippet: Up-regulation of CXCR3 ligand chemokines by TNFα in BM-MSCs. A , BM-MSCs were stimulated with TNFα (10 ng/ml) for various lengths of time (6–24 h). Total RNAs were prepared, and Northern blotting was performed. GAPDH expression

    Techniques Used: Northern Blot, Expressing

    Involvement of NF-κB in TNFα-induced CXCR3 ligand chemokines expression in BM-MSCs. A , BM-MSCs were treated with 10 ng/ml TNFα for the indicated times. Cytosol and nuclear fractions were isolated, and the amounts of IκB
    Figure Legend Snippet: Involvement of NF-κB in TNFα-induced CXCR3 ligand chemokines expression in BM-MSCs. A , BM-MSCs were treated with 10 ng/ml TNFα for the indicated times. Cytosol and nuclear fractions were isolated, and the amounts of IκB

    Techniques Used: Expressing, Isolation

    16) Product Images from "Transplantation of human bone marrow mesenchymal stromal cells reduces liver fibrosis more effectively than Wharton’s jelly mesenchymal stromal cells"

    Article Title: Transplantation of human bone marrow mesenchymal stromal cells reduces liver fibrosis more effectively than Wharton’s jelly mesenchymal stromal cells

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-017-0595-1

    Immunofluorescence analysis for a – c BM-MSCs and d – f WJ-MSCs engraftment in the liver tissues of CCl 4 -treated rats after 30 days of cell injection. Representative fluorescence images show colocalization of human CD105 expression ( a , d ) and DiI-positive cells ( b , e ) in liver tissue sections. The photomicrographs were captured using 40× and 60× (insets) objectives. BM-MSCs; bone marrow-derived mesenchymal stromal cells, WJ-MSCs; Wharton’s jelly-derived mesenchymal stromal cells
    Figure Legend Snippet: Immunofluorescence analysis for a – c BM-MSCs and d – f WJ-MSCs engraftment in the liver tissues of CCl 4 -treated rats after 30 days of cell injection. Representative fluorescence images show colocalization of human CD105 expression ( a , d ) and DiI-positive cells ( b , e ) in liver tissue sections. The photomicrographs were captured using 40× and 60× (insets) objectives. BM-MSCs; bone marrow-derived mesenchymal stromal cells, WJ-MSCs; Wharton’s jelly-derived mesenchymal stromal cells

    Techniques Used: Immunofluorescence, Injection, Fluorescence, Expressing, Derivative Assay

    17) Product Images from "Knockdown of insulin-like growth factor 1 exerts a protective effect on hypoxic injury of aged BM-MSCs: role of autophagy"

    Article Title: Knockdown of insulin-like growth factor 1 exerts a protective effect on hypoxic injury of aged BM-MSCs: role of autophagy

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-1028-5

    IGF-1 knockdown protects aged BM-MSCs by increasing autophagy. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA). b Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA) under normal and hypoxic conditions. c Quantification of apoptotic BM-MSCs under normal and hypoxic conditions. d Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in BM-MSCs of each group (scale bars, 20 mm). e Histogram showing the percentages of BM-MSCs with punctate LC3 in each group under normal and hypoxic conditions
    Figure Legend Snippet: IGF-1 knockdown protects aged BM-MSCs by increasing autophagy. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA). b Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA) under normal and hypoxic conditions. c Quantification of apoptotic BM-MSCs under normal and hypoxic conditions. d Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in BM-MSCs of each group (scale bars, 20 mm). e Histogram showing the percentages of BM-MSCs with punctate LC3 in each group under normal and hypoxic conditions

    Techniques Used: End Labeling, TUNEL Assay, Inhibition, Small Interfering RNA, Immunofluorescence, Fluorescence

    IGF-1 knockdown decreased apoptosis. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of BM-MSCs treated with H/SD with or without IGF-1 siRNA (scale bars, 20 μm). b The quantification result of the apoptotic BM-MSCs with or without IGF-1 siRNA. c Representative results of the FACS analysis of BM-MSCs under normal and H/SD conditions with or without IGF-1 siRNA administration. d Quantification of apoptosis is presented as the percentage of cells with the annexin marker in the early and late apoptotic stages with or without IGF-1 siRNA. Data are expressed as the means ± SEM; n = 5; * p
    Figure Legend Snippet: IGF-1 knockdown decreased apoptosis. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of BM-MSCs treated with H/SD with or without IGF-1 siRNA (scale bars, 20 μm). b The quantification result of the apoptotic BM-MSCs with or without IGF-1 siRNA. c Representative results of the FACS analysis of BM-MSCs under normal and H/SD conditions with or without IGF-1 siRNA administration. d Quantification of apoptosis is presented as the percentage of cells with the annexin marker in the early and late apoptotic stages with or without IGF-1 siRNA. Data are expressed as the means ± SEM; n = 5; * p

    Techniques Used: End Labeling, TUNEL Assay, FACS, Marker

    18) Product Images from "Cordycepin prevents oxidative stress-induced inhibition of osteogenesis"

    Article Title: Cordycepin prevents oxidative stress-induced inhibition of osteogenesis

    Journal: Oncotarget

    doi:

    Osteogenic differentiation of human BM-MSCs treated by 0.2 mM H 2 O 2 , 10 μg/mL cordycepin and 0.2 mM H 2 O 2 +10 μg/mL cordycepin co-treatment Relative OPN , Collagen I mRNA expression a. and OPG and RANKL mRNA expression in the experimental groups were characterized by RT-PCR analysis. Gene expression was normalized to GAPDH . Data were shown as mean ± S.E.M. * p
    Figure Legend Snippet: Osteogenic differentiation of human BM-MSCs treated by 0.2 mM H 2 O 2 , 10 μg/mL cordycepin and 0.2 mM H 2 O 2 +10 μg/mL cordycepin co-treatment Relative OPN , Collagen I mRNA expression a. and OPG and RANKL mRNA expression in the experimental groups were characterized by RT-PCR analysis. Gene expression was normalized to GAPDH . Data were shown as mean ± S.E.M. * p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    19) Product Images from "Knockdown of insulin-like growth factor 1 exerts a protective effect on hypoxic injury of aged BM-MSCs: role of autophagy"

    Article Title: Knockdown of insulin-like growth factor 1 exerts a protective effect on hypoxic injury of aged BM-MSCs: role of autophagy

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-1028-5

    Proposed mechanism for how IGF-1 knockdown protects aged BM-MSCs against hypoxic injury. In young BM-MSCs, the appropriate amount of IGF-1 bound to its receptor promotes mTOR and Akt phosphorylation (activation of mTOR/Akt), thereby activating S6K and 4EBP1, which leads to the maintenance of basal autophagy. Basal autophagy maintains cell survival and thus protects the heart. In aged BM-MSCs, there is excessive IGF-1 bound to its receptor, which leads to the overactivation of mTOR/Akt, S6K, and 4EBP1; therefore, the inhibition of basal autophagy is overly promoted, which accelerates apoptosis in aged BM-MSCs. As a result, insufficient autophagy can maintain neither cell functional survival nor protect the heart
    Figure Legend Snippet: Proposed mechanism for how IGF-1 knockdown protects aged BM-MSCs against hypoxic injury. In young BM-MSCs, the appropriate amount of IGF-1 bound to its receptor promotes mTOR and Akt phosphorylation (activation of mTOR/Akt), thereby activating S6K and 4EBP1, which leads to the maintenance of basal autophagy. Basal autophagy maintains cell survival and thus protects the heart. In aged BM-MSCs, there is excessive IGF-1 bound to its receptor, which leads to the overactivation of mTOR/Akt, S6K, and 4EBP1; therefore, the inhibition of basal autophagy is overly promoted, which accelerates apoptosis in aged BM-MSCs. As a result, insufficient autophagy can maintain neither cell functional survival nor protect the heart

    Techniques Used: Activation Assay, Inhibition, Functional Assay

    IGF-1 knockdown decreased the activity of the Akt/mTOR signaling pathway in aged BM-MSCs. a Representative western blots of p-Akt/Akt and p-mTOR/mTOR in aged BM-MSCs. b Representative western blots of p-p70S6, K/p70S6 K, and p-S6/S6 in aged BM-MSCs. Semiquantification of the protein expression levels of p-Akt ( c ), p-mTOR ( d ) ( n = 5, * p
    Figure Legend Snippet: IGF-1 knockdown decreased the activity of the Akt/mTOR signaling pathway in aged BM-MSCs. a Representative western blots of p-Akt/Akt and p-mTOR/mTOR in aged BM-MSCs. b Representative western blots of p-p70S6, K/p70S6 K, and p-S6/S6 in aged BM-MSCs. Semiquantification of the protein expression levels of p-Akt ( c ), p-mTOR ( d ) ( n = 5, * p

    Techniques Used: Activity Assay, Western Blot, Expressing

    IGF-1 knockdown protects aged BM-MSCs by increasing autophagy. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA). b Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA) under normal and hypoxic conditions. c Quantification of apoptotic BM-MSCs under normal and hypoxic conditions. d Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in BM-MSCs of each group (scale bars, 20 mm). e Histogram showing the percentages of BM-MSCs with punctate LC3 in each group under normal and hypoxic conditions
    Figure Legend Snippet: IGF-1 knockdown protects aged BM-MSCs by increasing autophagy. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA). b Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA) under normal and hypoxic conditions. c Quantification of apoptotic BM-MSCs under normal and hypoxic conditions. d Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in BM-MSCs of each group (scale bars, 20 mm). e Histogram showing the percentages of BM-MSCs with punctate LC3 in each group under normal and hypoxic conditions

    Techniques Used: End Labeling, TUNEL Assay, Inhibition, Small Interfering RNA, Immunofluorescence, Fluorescence

    IGF-1 knockdown decreased apoptosis. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of BM-MSCs treated with H/SD with or without IGF-1 siRNA (scale bars, 20 μm). b The quantification result of the apoptotic BM-MSCs with or without IGF-1 siRNA. c Representative results of the FACS analysis of BM-MSCs under normal and H/SD conditions with or without IGF-1 siRNA administration. d Quantification of apoptosis is presented as the percentage of cells with the annexin marker in the early and late apoptotic stages with or without IGF-1 siRNA. Data are expressed as the means ± SEM; n = 5; * p
    Figure Legend Snippet: IGF-1 knockdown decreased apoptosis. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of BM-MSCs treated with H/SD with or without IGF-1 siRNA (scale bars, 20 μm). b The quantification result of the apoptotic BM-MSCs with or without IGF-1 siRNA. c Representative results of the FACS analysis of BM-MSCs under normal and H/SD conditions with or without IGF-1 siRNA administration. d Quantification of apoptosis is presented as the percentage of cells with the annexin marker in the early and late apoptotic stages with or without IGF-1 siRNA. Data are expressed as the means ± SEM; n = 5; * p

    Techniques Used: End Labeling, TUNEL Assay, FACS, Marker

    IGF-1 knockdown increased autophagy. a Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in the BM-MSCs of each group. b Quantification of autophagy is shown as the percentage of BM-MSCs with LC3 ( n = 5, * p
    Figure Legend Snippet: IGF-1 knockdown increased autophagy. a Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in the BM-MSCs of each group. b Quantification of autophagy is shown as the percentage of BM-MSCs with LC3 ( n = 5, * p

    Techniques Used: Immunofluorescence, Fluorescence

    20) Product Images from "Anterior cruciate ligament-derived mesenchymal stromal cells have a propensity to differentiate into the ligament lineage"

    Article Title: Anterior cruciate ligament-derived mesenchymal stromal cells have a propensity to differentiate into the ligament lineage

    Journal: Regenerative Therapy

    doi: 10.1016/j.reth.2017.12.001

    Ability to differentiate into adipocytes, osteoblasts, and ligament cells (a, b) Representative phase contrast micrographs of ACL- and BM-MSCs differentiated into adipocytes (a) and osteoblasts (b) (Scale bars = 100 μm). (c) Expression levels of mRNA ( MKX , SCX , and COL1A1 ) following ligament differentiation (n = 5, p**
    Figure Legend Snippet: Ability to differentiate into adipocytes, osteoblasts, and ligament cells (a, b) Representative phase contrast micrographs of ACL- and BM-MSCs differentiated into adipocytes (a) and osteoblasts (b) (Scale bars = 100 μm). (c) Expression levels of mRNA ( MKX , SCX , and COL1A1 ) following ligament differentiation (n = 5, p**

    Techniques Used: Expressing

    Comparison of cell surface proteins between ACL- and bone marrow (BM)-derived MSCs. Cell surface protein expression in ACL-MSCs (CD73 + /90 + ) and BM-MSCs (CD90 + /271 + ) before (a) and after culture (b). (c) Principal component analysis of four types of cells. ACL-derived CD73 + /90 + MSCs: fresh (blue, circle) and cultured (blue, square); BM-derived CD90 + /271 + MSCs: fresh (red, circle) and cultured (red, square).
    Figure Legend Snippet: Comparison of cell surface proteins between ACL- and bone marrow (BM)-derived MSCs. Cell surface protein expression in ACL-MSCs (CD73 + /90 + ) and BM-MSCs (CD90 + /271 + ) before (a) and after culture (b). (c) Principal component analysis of four types of cells. ACL-derived CD73 + /90 + MSCs: fresh (blue, circle) and cultured (blue, square); BM-derived CD90 + /271 + MSCs: fresh (red, circle) and cultured (red, square).

    Techniques Used: Derivative Assay, Expressing, Cell Culture

    21) Product Images from "H2O2 Accumulation Mediates Differentiation Capacity Alteration, But Not Proliferative Decline, in Senescent Human Fetal Mesenchymal Stem Cells"

    Article Title: H2O2 Accumulation Mediates Differentiation Capacity Alteration, But Not Proliferative Decline, in Senescent Human Fetal Mesenchymal Stem Cells

    Journal: Antioxidants & Redox Signaling

    doi: 10.1089/ars.2012.4692

    PDMCs become senescent after prolonged in vitro culture. (A) In vitro proliferation of PDMCs (three donors #1–3) and BM-MSCs (two donors #1–2). (B) Doubling times of PDMCs (#1–3) and BM-MSCs (#1–2) within a different range
    Figure Legend Snippet: PDMCs become senescent after prolonged in vitro culture. (A) In vitro proliferation of PDMCs (three donors #1–3) and BM-MSCs (two donors #1–2). (B) Doubling times of PDMCs (#1–3) and BM-MSCs (#1–2) within a different range

    Techniques Used: In Vitro

    22) Product Images from "Sphingosine 1-Phosphate Receptor 1 Is Required for MMP-2 Function in Bone Marrow Mesenchymal Stromal Cells: Implications for Cytoskeleton Assembly and Proliferation"

    Article Title: Sphingosine 1-Phosphate Receptor 1 Is Required for MMP-2 Function in Bone Marrow Mesenchymal Stromal Cells: Implications for Cytoskeleton Assembly and Proliferation

    Journal: Stem Cells International

    doi: 10.1155/2018/5034679

    MMP-2 expression and activity. BM-MSCs were cultured for 48 h in the absence (vehicle) or in presence of 1 μ M exogenous sphingosine-1-phosphate (exoS1P) or 2 μ M S1PR1 receptor antagonist, W146, or 2 μ M S1PR1 receptor agonist, SEW2871. (a) Representative immunofluorescence confocal images of fixed cells on glass coverslips immunostained with antibodies against MMP-2 (green). Scale bar 50 μ m. The images are representative of at least three independent experiments with similar results. (b) Densitometric analysis of the intensity of the MMP-2 fluorescence signal performed on digitized images. (c) Zymography. A representative gelatin zymography of MMP-2 from conditioned media obtained from BM-MSCs incubated in absence (vehicle) or in presence of W146 or SEW2871 for 48 h. Densitometry scanning from at least three separate experiments was performed and data, expressed as relative OD values (a.u.) to those of control group (vehicle) set to 100, are reported in the histogram. Data are mean ± S.E.M. Significance of difference in (b) (one-way ANOVA and Newman-Keuls multiple comparison tests), ∗ p
    Figure Legend Snippet: MMP-2 expression and activity. BM-MSCs were cultured for 48 h in the absence (vehicle) or in presence of 1 μ M exogenous sphingosine-1-phosphate (exoS1P) or 2 μ M S1PR1 receptor antagonist, W146, or 2 μ M S1PR1 receptor agonist, SEW2871. (a) Representative immunofluorescence confocal images of fixed cells on glass coverslips immunostained with antibodies against MMP-2 (green). Scale bar 50 μ m. The images are representative of at least three independent experiments with similar results. (b) Densitometric analysis of the intensity of the MMP-2 fluorescence signal performed on digitized images. (c) Zymography. A representative gelatin zymography of MMP-2 from conditioned media obtained from BM-MSCs incubated in absence (vehicle) or in presence of W146 or SEW2871 for 48 h. Densitometry scanning from at least three separate experiments was performed and data, expressed as relative OD values (a.u.) to those of control group (vehicle) set to 100, are reported in the histogram. Data are mean ± S.E.M. Significance of difference in (b) (one-way ANOVA and Newman-Keuls multiple comparison tests), ∗ p

    Techniques Used: Expressing, Activity Assay, Cell Culture, Immunofluorescence, Fluorescence, Zymography, Incubation

    Cytoskeleton organization and cortactin expression. BM-MSCs were cultured for 48 h in the absence (vehicle) or in presence of the following compounds: 1 μ M exogenous sphingosine-1-phosphate (exoS1P), 2 μ M S1PR1 receptor antagonist, W146, and 2 μ M S1PR1 receptor agonist, SEW2871 and/or MMP-2/9 inhibitor, SB-3CT (5 μ M or 10 μ M). (a–h) Representative immunofluorescence confocal images of cells cultured on glass coverslips in the indicated experimental conditions, fixed and stained with Alexa 568-phalloidin to visualize actin filaments (red) and immunostained with antibodies against cortactin (green). Scale bar 50 μ m. Arrows indicate filopodia and arrowheads indicate lamellipodia (L). (A–D) Magnifications of the indicated squared regions of interest showing the red and green fluorescence signals separately and together. Yellow-orange colour indicates colocalization between the two fluorescence signals. Scale bar 12 μ m. The images are representative of at least three independent experiments with similar results. (i) Densitometric analysis of the intensity of the cortactin fluorescence signal performed on digitized images. Data are mean ± S.E.M. Significance of differences (one-way ANOVA and Newman-Keuls multiple comparison test): ∗ p
    Figure Legend Snippet: Cytoskeleton organization and cortactin expression. BM-MSCs were cultured for 48 h in the absence (vehicle) or in presence of the following compounds: 1 μ M exogenous sphingosine-1-phosphate (exoS1P), 2 μ M S1PR1 receptor antagonist, W146, and 2 μ M S1PR1 receptor agonist, SEW2871 and/or MMP-2/9 inhibitor, SB-3CT (5 μ M or 10 μ M). (a–h) Representative immunofluorescence confocal images of cells cultured on glass coverslips in the indicated experimental conditions, fixed and stained with Alexa 568-phalloidin to visualize actin filaments (red) and immunostained with antibodies against cortactin (green). Scale bar 50 μ m. Arrows indicate filopodia and arrowheads indicate lamellipodia (L). (A–D) Magnifications of the indicated squared regions of interest showing the red and green fluorescence signals separately and together. Yellow-orange colour indicates colocalization between the two fluorescence signals. Scale bar 12 μ m. The images are representative of at least three independent experiments with similar results. (i) Densitometric analysis of the intensity of the cortactin fluorescence signal performed on digitized images. Data are mean ± S.E.M. Significance of differences (one-way ANOVA and Newman-Keuls multiple comparison test): ∗ p

    Techniques Used: Expressing, Cell Culture, Immunofluorescence, Staining, Fluorescence

    S1P receptor subtype expression and SphK/S1PR axis role in cell gelatinolytic activity. (a and c) Expression of S1P receptors by reverse transcription (RT) and real-time PCR analysis. mRNA were determined by RT of total RNA (1 μ g) obtained from BM-MSCs at low- (L-) and high- (H-) density culture and 2 μ l of cDNA (for S1PR1, S1PR2, and S1PR3 detection) or 4 μ . Representative agarose gels of amplified DNA are shown. GAPDH amplification was used for data normalization. (b and d) Quantification of mRNA expression by real-time PCR analysis. Data are reported as mean ± S.E.M. of the ratio between the fold of variation of S1P receptor expression obtained from high- and low-density BM-MSCs culture. (e and f) BM-MSCs seeded onto fluorescein-labeled gelatin substrate- (DQ gelatin-) coated plastic culture plates (e) or glass coverslips (f) were cultured for 48 h in absence (vehicle) or in presence of the following compounds: 5 μ M sphingosine kinase inhibitor (iSK), 1 μ M exogenous sphingosine-1-phosphate (exoS1P), 2 μ M S1PR1 receptor antagonist, W146, and 2 μ M S1PR1 receptor agonist, SEW2871. (e) Spectrophotometrical quantification of the DQ gelatin fluorescence intensity revealed after proteolytic digestion of the gelatin by MMP gelatinases. (f) Representative superimposed DIC (grey) and fluorescent confocal microscopy images (green; gelatin fluorescence intensity) of fixed cells. Scale bar 30 μ m. Histogram shows the densitometric analysis of the intensity of the gelatin fluorescence signals performed on digitized images. Data reported as mean ± S.E.M. are representative of at least three independent experiments with similar results. Significance of differences in (b) and (d) (Student's t -test), ∗ p
    Figure Legend Snippet: S1P receptor subtype expression and SphK/S1PR axis role in cell gelatinolytic activity. (a and c) Expression of S1P receptors by reverse transcription (RT) and real-time PCR analysis. mRNA were determined by RT of total RNA (1 μ g) obtained from BM-MSCs at low- (L-) and high- (H-) density culture and 2 μ l of cDNA (for S1PR1, S1PR2, and S1PR3 detection) or 4 μ . Representative agarose gels of amplified DNA are shown. GAPDH amplification was used for data normalization. (b and d) Quantification of mRNA expression by real-time PCR analysis. Data are reported as mean ± S.E.M. of the ratio between the fold of variation of S1P receptor expression obtained from high- and low-density BM-MSCs culture. (e and f) BM-MSCs seeded onto fluorescein-labeled gelatin substrate- (DQ gelatin-) coated plastic culture plates (e) or glass coverslips (f) were cultured for 48 h in absence (vehicle) or in presence of the following compounds: 5 μ M sphingosine kinase inhibitor (iSK), 1 μ M exogenous sphingosine-1-phosphate (exoS1P), 2 μ M S1PR1 receptor antagonist, W146, and 2 μ M S1PR1 receptor agonist, SEW2871. (e) Spectrophotometrical quantification of the DQ gelatin fluorescence intensity revealed after proteolytic digestion of the gelatin by MMP gelatinases. (f) Representative superimposed DIC (grey) and fluorescent confocal microscopy images (green; gelatin fluorescence intensity) of fixed cells. Scale bar 30 μ m. Histogram shows the densitometric analysis of the intensity of the gelatin fluorescence signals performed on digitized images. Data reported as mean ± S.E.M. are representative of at least three independent experiments with similar results. Significance of differences in (b) and (d) (Student's t -test), ∗ p

    Techniques Used: Expressing, Activity Assay, Real-time Polymerase Chain Reaction, Amplification, Labeling, Cell Culture, Fluorescence, Confocal Microscopy

    MMP-2 expression and activity in MSCs cultured under hypoxic conditions. BM-MSCs were cultured for 48 h under hypoxic conditions in the absence (vehicle) or in presence of 2 μ M S1PR1 receptor antagonist, W146, or 2 μ M S1PR1 receptor agonist, SEW2871. (a) Representative immunofluorescence confocal images of cells cultured on glass coverslips, fixed and immunostained with antibodies against MMP-2 (green). Scale bar 50 μ m. The images are representative of at least three independent experiments with similar results. (b) Densitometric analysis of the intensity of the MMP-2 fluorescence signal performed on digitized images. Data are mean ± S.E.M. (c) Zymography. A representative gelatin zymography of MMP-2 from conditioned media obtained from BM-MSCs incubated in the absence (vehicle) or in the presence of 2 μ M S1PR1 receptor antagonist, W146, or 2 μ M S1PR1 receptor agonist, SEW2871, for 48 h. Densitometry scanning from at least three separate experiments was performed and data, expressed as relative OD values (a.u.) to those of the control group (vehicle) set to 100, are reported in the histogram. Data are mean ± S.E.M. Significance of differences in (b) and (c) (Student's t -test): ∗ p
    Figure Legend Snippet: MMP-2 expression and activity in MSCs cultured under hypoxic conditions. BM-MSCs were cultured for 48 h under hypoxic conditions in the absence (vehicle) or in presence of 2 μ M S1PR1 receptor antagonist, W146, or 2 μ M S1PR1 receptor agonist, SEW2871. (a) Representative immunofluorescence confocal images of cells cultured on glass coverslips, fixed and immunostained with antibodies against MMP-2 (green). Scale bar 50 μ m. The images are representative of at least three independent experiments with similar results. (b) Densitometric analysis of the intensity of the MMP-2 fluorescence signal performed on digitized images. Data are mean ± S.E.M. (c) Zymography. A representative gelatin zymography of MMP-2 from conditioned media obtained from BM-MSCs incubated in the absence (vehicle) or in the presence of 2 μ M S1PR1 receptor antagonist, W146, or 2 μ M S1PR1 receptor agonist, SEW2871, for 48 h. Densitometry scanning from at least three separate experiments was performed and data, expressed as relative OD values (a.u.) to those of the control group (vehicle) set to 100, are reported in the histogram. Data are mean ± S.E.M. Significance of differences in (b) and (c) (Student's t -test): ∗ p

    Techniques Used: Expressing, Activity Assay, Cell Culture, Immunofluorescence, Fluorescence, Zymography, Incubation

    23) Product Images from "Isolation of alveolar epithelial type II progenitor cells from adult human lungs"

    Article Title: Isolation of alveolar epithelial type II progenitor cells from adult human lungs

    Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

    doi: 10.1038/labinvest.2010.187

    Comparison of transcription profiles between pro-SP-C + /CD90 + cells and bone marrow-mesenchymal stem cells (BM-MSCs). ( a ) The area-proportional Venn diagram presenting the overlap between transcripts expressed in pro-SP-C + /CD90 + cells (blue) and BM-MSCs (red). ( b ) Functional annotation clustering in specifically expressed gene sets in pro-SP-C + /CD90 + cells (blue) or BM-MSCs (red). Original array data are available at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=zpapfyaqumsqmvw acc=GSE21095 . ( c ) Semiquantitative RT–PCR for eight genes identified through the microarray analysis. Forkhead box f1 (Foxf1), T-box 4 (Tbx4), FBJ murine osteosarcoma viral oncogene homolog B (FosB), and laminin α 5, that were selected out of the annotations of ‘transcription' and/or ‘lung development', were highly expressed by pro-SP-C + /CD90 + cells. At the same time, distal-less homeobox 5 (DLX5), N-cadherin, homeobox C10 (HOXC10), and hyaluronan synthase 1 (HAS1), that were chosen from the annotation of ‘skeletal system development' or ‘cell adhesion', were highly expressed by BM-MSCs. Three bathes of pro-SP-C + /CD90 + cells and three batches of BM-MSCs were examined. β -Actin was used as an endogenous control. In RT–PCR for Foxf1, Tbx4, FosB, laminin α 5, and β -actin, a representative negative control (pro-SP-C + /CD90 + cells batch1 without reverse transcriptase reaction) is shown as no-RT. In RT–PCR for DLX5, N-cadherin, HOXC10, and HAS1, a representative negative control (BM-MSCs batch1 without reverse transcriptase reaction) is shown as no-RT.
    Figure Legend Snippet: Comparison of transcription profiles between pro-SP-C + /CD90 + cells and bone marrow-mesenchymal stem cells (BM-MSCs). ( a ) The area-proportional Venn diagram presenting the overlap between transcripts expressed in pro-SP-C + /CD90 + cells (blue) and BM-MSCs (red). ( b ) Functional annotation clustering in specifically expressed gene sets in pro-SP-C + /CD90 + cells (blue) or BM-MSCs (red). Original array data are available at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=zpapfyaqumsqmvw acc=GSE21095 . ( c ) Semiquantitative RT–PCR for eight genes identified through the microarray analysis. Forkhead box f1 (Foxf1), T-box 4 (Tbx4), FBJ murine osteosarcoma viral oncogene homolog B (FosB), and laminin α 5, that were selected out of the annotations of ‘transcription' and/or ‘lung development', were highly expressed by pro-SP-C + /CD90 + cells. At the same time, distal-less homeobox 5 (DLX5), N-cadherin, homeobox C10 (HOXC10), and hyaluronan synthase 1 (HAS1), that were chosen from the annotation of ‘skeletal system development' or ‘cell adhesion', were highly expressed by BM-MSCs. Three bathes of pro-SP-C + /CD90 + cells and three batches of BM-MSCs were examined. β -Actin was used as an endogenous control. In RT–PCR for Foxf1, Tbx4, FosB, laminin α 5, and β -actin, a representative negative control (pro-SP-C + /CD90 + cells batch1 without reverse transcriptase reaction) is shown as no-RT. In RT–PCR for DLX5, N-cadherin, HOXC10, and HAS1, a representative negative control (BM-MSCs batch1 without reverse transcriptase reaction) is shown as no-RT.

    Techniques Used: Functional Assay, Reverse Transcription Polymerase Chain Reaction, Microarray, Negative Control

    24) Product Images from "Anterior cruciate ligament-derived mesenchymal stromal cells have a propensity to differentiate into the ligament lineage"

    Article Title: Anterior cruciate ligament-derived mesenchymal stromal cells have a propensity to differentiate into the ligament lineage

    Journal: Regenerative Therapy

    doi: 10.1016/j.reth.2017.12.001

    Ability to differentiate into adipocytes, osteoblasts, and ligament cells (a, b) Representative phase contrast micrographs of ACL- and BM-MSCs differentiated into adipocytes (a) and osteoblasts (b) (Scale bars = 100 μm). (c) Expression levels of mRNA ( MKX , SCX , and COL1A1 ) following ligament differentiation (n = 5, p**
    Figure Legend Snippet: Ability to differentiate into adipocytes, osteoblasts, and ligament cells (a, b) Representative phase contrast micrographs of ACL- and BM-MSCs differentiated into adipocytes (a) and osteoblasts (b) (Scale bars = 100 μm). (c) Expression levels of mRNA ( MKX , SCX , and COL1A1 ) following ligament differentiation (n = 5, p**

    Techniques Used: Expressing

    Comparison of cell surface proteins between ACL- and bone marrow (BM)-derived MSCs. Cell surface protein expression in ACL-MSCs (CD73 + /90 + ) and BM-MSCs (CD90 + /271 + ) before (a) and after culture (b). (c) Principal component analysis of four types of cells. ACL-derived CD73 + /90 + MSCs: fresh (blue, circle) and cultured (blue, square); BM-derived CD90 + /271 + MSCs: fresh (red, circle) and cultured (red, square).
    Figure Legend Snippet: Comparison of cell surface proteins between ACL- and bone marrow (BM)-derived MSCs. Cell surface protein expression in ACL-MSCs (CD73 + /90 + ) and BM-MSCs (CD90 + /271 + ) before (a) and after culture (b). (c) Principal component analysis of four types of cells. ACL-derived CD73 + /90 + MSCs: fresh (blue, circle) and cultured (blue, square); BM-derived CD90 + /271 + MSCs: fresh (red, circle) and cultured (red, square).

    Techniques Used: Derivative Assay, Expressing, Cell Culture

    25) Product Images from "Exosomes as potential alternatives to stem cell therapy for intervertebral disc degeneration: in-vitro study on exosomes in interaction of nucleus pulposus cells and bone marrow mesenchymal stem cells"

    Article Title: Exosomes as potential alternatives to stem cell therapy for intervertebral disc degeneration: in-vitro study on exosomes in interaction of nucleus pulposus cells and bone marrow mesenchymal stem cells

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-017-0563-9

    Characterization of BM-MSC and NPC-derived exosomes by TEM. a PBS used as a control group; b exosomes derived from BM-MSCs; c exosomes derived from NPCs. Exosomes were stained with phosphotungstic acid and observed by TEM (×100,000). The two kinds of exosomes were roughly identical, ranging from 30 to 100 nm. Scale bar = 200 nm. BM-MSC bone marrow mesenchymal stem cell, Exo exosomes, NPC nucleus pulposus cell, PBS phosphate-buffered saline
    Figure Legend Snippet: Characterization of BM-MSC and NPC-derived exosomes by TEM. a PBS used as a control group; b exosomes derived from BM-MSCs; c exosomes derived from NPCs. Exosomes were stained with phosphotungstic acid and observed by TEM (×100,000). The two kinds of exosomes were roughly identical, ranging from 30 to 100 nm. Scale bar = 200 nm. BM-MSC bone marrow mesenchymal stem cell, Exo exosomes, NPC nucleus pulposus cell, PBS phosphate-buffered saline

    Techniques Used: Derivative Assay, Transmission Electron Microscopy, Staining

    26) Product Images from "Knockdown of insulin-like growth factor 1 exerts a protective effect on hypoxic injury of aged BM-MSCs: role of autophagy"

    Article Title: Knockdown of insulin-like growth factor 1 exerts a protective effect on hypoxic injury of aged BM-MSCs: role of autophagy

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-1028-5

    Proposed mechanism for how IGF-1 knockdown protects aged BM-MSCs against hypoxic injury. In young BM-MSCs, the appropriate amount of IGF-1 bound to its receptor promotes mTOR and Akt phosphorylation (activation of mTOR/Akt), thereby activating S6K and 4EBP1, which leads to the maintenance of basal autophagy. Basal autophagy maintains cell survival and thus protects the heart. In aged BM-MSCs, there is excessive IGF-1 bound to its receptor, which leads to the overactivation of mTOR/Akt, S6K, and 4EBP1; therefore, the inhibition of basal autophagy is overly promoted, which accelerates apoptosis in aged BM-MSCs. As a result, insufficient autophagy can maintain neither cell functional survival nor protect the heart
    Figure Legend Snippet: Proposed mechanism for how IGF-1 knockdown protects aged BM-MSCs against hypoxic injury. In young BM-MSCs, the appropriate amount of IGF-1 bound to its receptor promotes mTOR and Akt phosphorylation (activation of mTOR/Akt), thereby activating S6K and 4EBP1, which leads to the maintenance of basal autophagy. Basal autophagy maintains cell survival and thus protects the heart. In aged BM-MSCs, there is excessive IGF-1 bound to its receptor, which leads to the overactivation of mTOR/Akt, S6K, and 4EBP1; therefore, the inhibition of basal autophagy is overly promoted, which accelerates apoptosis in aged BM-MSCs. As a result, insufficient autophagy can maintain neither cell functional survival nor protect the heart

    Techniques Used: Activation Assay, Inhibition, Functional Assay

    IGF-1 knockdown decreased the activity of the Akt/mTOR signaling pathway in aged BM-MSCs. a Representative western blots of p-Akt/Akt and p-mTOR/mTOR in aged BM-MSCs. b Representative western blots of p-p70S6, K/p70S6 K, and p-S6/S6 in aged BM-MSCs. Semiquantification of the protein expression levels of p-Akt ( c ), p-mTOR ( d ) ( n = 5, * p
    Figure Legend Snippet: IGF-1 knockdown decreased the activity of the Akt/mTOR signaling pathway in aged BM-MSCs. a Representative western blots of p-Akt/Akt and p-mTOR/mTOR in aged BM-MSCs. b Representative western blots of p-p70S6, K/p70S6 K, and p-S6/S6 in aged BM-MSCs. Semiquantification of the protein expression levels of p-Akt ( c ), p-mTOR ( d ) ( n = 5, * p

    Techniques Used: Activity Assay, Western Blot, Expressing

    IGF-1 knockdown protects aged BM-MSCs by increasing autophagy. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA). b Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA) under normal and hypoxic conditions. c Quantification of apoptotic BM-MSCs under normal and hypoxic conditions. d Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in BM-MSCs of each group (scale bars, 20 mm). e Histogram showing the percentages of BM-MSCs with punctate LC3 in each group under normal and hypoxic conditions
    Figure Legend Snippet: IGF-1 knockdown protects aged BM-MSCs by increasing autophagy. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA). b Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of aged BM-MSCs treated with IGF-1 siRNA with or without autophagy inhibition via 3-methyladenine (3-MA) and Atg7 small interfering RNA (siRNA) under normal and hypoxic conditions. c Quantification of apoptotic BM-MSCs under normal and hypoxic conditions. d Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in BM-MSCs of each group (scale bars, 20 mm). e Histogram showing the percentages of BM-MSCs with punctate LC3 in each group under normal and hypoxic conditions

    Techniques Used: End Labeling, TUNEL Assay, Inhibition, Small Interfering RNA, Immunofluorescence, Fluorescence

    IGF-1 knockdown decreased apoptosis. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of BM-MSCs treated with H/SD with or without IGF-1 siRNA (scale bars, 20 μm). b The quantification result of the apoptotic BM-MSCs with or without IGF-1 siRNA. c Representative results of the FACS analysis of BM-MSCs under normal and H/SD conditions with or without IGF-1 siRNA administration. d Quantification of apoptosis is presented as the percentage of cells with the annexin marker in the early and late apoptotic stages with or without IGF-1 siRNA. Data are expressed as the means ± SEM; n = 5; * p
    Figure Legend Snippet: IGF-1 knockdown decreased apoptosis. a Representative terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) images of BM-MSCs treated with H/SD with or without IGF-1 siRNA (scale bars, 20 μm). b The quantification result of the apoptotic BM-MSCs with or without IGF-1 siRNA. c Representative results of the FACS analysis of BM-MSCs under normal and H/SD conditions with or without IGF-1 siRNA administration. d Quantification of apoptosis is presented as the percentage of cells with the annexin marker in the early and late apoptotic stages with or without IGF-1 siRNA. Data are expressed as the means ± SEM; n = 5; * p

    Techniques Used: End Labeling, TUNEL Assay, FACS, Marker

    IGF-1 knockdown increased autophagy. a Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in the BM-MSCs of each group. b Quantification of autophagy is shown as the percentage of BM-MSCs with LC3 ( n = 5, * p
    Figure Legend Snippet: IGF-1 knockdown increased autophagy. a Representative immunofluorescence images of GFP-LC3 (green fluorescent) and DAPI (blue fluorescence) in the BM-MSCs of each group. b Quantification of autophagy is shown as the percentage of BM-MSCs with LC3 ( n = 5, * p

    Techniques Used: Immunofluorescence, Fluorescence

    27) Product Images from "TNF?-exposed Bone Marrow-derived Mesenchymal Stem Cells Promote Locomotion of MDA-MB-231 Breast Cancer Cells through Transcriptional Activation of CXCR3 Ligand Chemokines *"

    Article Title: TNF?-exposed Bone Marrow-derived Mesenchymal Stem Cells Promote Locomotion of MDA-MB-231 Breast Cancer Cells through Transcriptional Activation of CXCR3 Ligand Chemokines *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.128124

    NF-κB binding site is essential for TNFα-induced promoter activation of CXCR3 ligand chemokines. BM-MSCs were transiently transfected with 0.2 μg of serial deletion or NF-κB site mutant ( mtNF κ B ) constructs of CXCL9
    Figure Legend Snippet: NF-κB binding site is essential for TNFα-induced promoter activation of CXCR3 ligand chemokines. BM-MSCs were transiently transfected with 0.2 μg of serial deletion or NF-κB site mutant ( mtNF κ B ) constructs of CXCL9

    Techniques Used: Binding Assay, Activation Assay, Transfection, Mutagenesis, Construct

    28) Product Images from "LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK"

    Article Title: LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK

    Journal: Stem Cells (Dayton, Ohio)

    doi: 10.1002/stem.2937

    Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p
    Figure Legend Snippet: Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p

    Techniques Used: Expressing, Over Expression, Derivative Assay, Western Blot, Staining, Activity Assay

    29) Product Images from "MiR-1908/EXO1 and MiR-203a/FOS, regulated by scd1, are associated with fracture risk and bone health in postmenopausal diabetic women"

    Article Title: MiR-1908/EXO1 and MiR-203a/FOS, regulated by scd1, are associated with fracture risk and bone health in postmenopausal diabetic women

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.103227

    SCD1 overexpression promotes osteogenic differentiation and induces expression changes in BM-MSCs. ( A ) Fluorescence staining showed the transfection of BM-MSCs with SCD1 lentivirus. Comparison results of SCD1 expression and activity between the SCD1 overexpression and control groups were showed in the table beneath. Data are expressed as means ± standard deviations ( n = 6). * P
    Figure Legend Snippet: SCD1 overexpression promotes osteogenic differentiation and induces expression changes in BM-MSCs. ( A ) Fluorescence staining showed the transfection of BM-MSCs with SCD1 lentivirus. Comparison results of SCD1 expression and activity between the SCD1 overexpression and control groups were showed in the table beneath. Data are expressed as means ± standard deviations ( n = 6). * P

    Techniques Used: Over Expression, Expressing, Fluorescence, Staining, Transfection, Activity Assay

    Correlation between miR-1908 and miR-203a expression levels and SCD1. Differential expression of miR-1908 and miR-203a after 3 days ( A , B ) and 1 week ( C ) of SCD1 overexpression in BM-MSCs. ( D , E ) showed the relative expression of miR-1908 and miR-203a detected by qPCR in patients with clinical diabetic fractures.
    Figure Legend Snippet: Correlation between miR-1908 and miR-203a expression levels and SCD1. Differential expression of miR-1908 and miR-203a after 3 days ( A , B ) and 1 week ( C ) of SCD1 overexpression in BM-MSCs. ( D , E ) showed the relative expression of miR-1908 and miR-203a detected by qPCR in patients with clinical diabetic fractures.

    Techniques Used: Expressing, Over Expression, Real-time Polymerase Chain Reaction

    miR-203a/FOS and miR-1908/EXO1 are regulated by SCD1. ( A ) Transfection efficiency of SCD1 in BM-MSCs. ( B ) Proliferation of BM-MSCs after SCD1 overexpression, miR-1908mimic and miR-203a inhibitor were evaluated. ( C ) ALP activity was measured in SCD1-overexpressing BM-MSCs treated with miR-203a inhibitor or miR-1908 mimic. ( D , E ) Expressions of FOS and EXO1 were assessed in BM-MSCs transfected with high-glucose, SCD-OE miR-1908 mimic and miR-203a inhibitor. ( F ) The regulatory network between dysregulated miRNAs and hub genes after overexpression SCD-1.
    Figure Legend Snippet: miR-203a/FOS and miR-1908/EXO1 are regulated by SCD1. ( A ) Transfection efficiency of SCD1 in BM-MSCs. ( B ) Proliferation of BM-MSCs after SCD1 overexpression, miR-1908mimic and miR-203a inhibitor were evaluated. ( C ) ALP activity was measured in SCD1-overexpressing BM-MSCs treated with miR-203a inhibitor or miR-1908 mimic. ( D , E ) Expressions of FOS and EXO1 were assessed in BM-MSCs transfected with high-glucose, SCD-OE miR-1908 mimic and miR-203a inhibitor. ( F ) The regulatory network between dysregulated miRNAs and hub genes after overexpression SCD-1.

    Techniques Used: Transfection, Over Expression, Activity Assay

    CDKN1A, FOS, EXO1 and PLS1 are regulated by SCD1. ( A ) Independent t -test results for the association between mRNAs (EXO1, PLS1, FOS and CDKN1A) and trauma energy in patients with clinical diabetic fractures patients. ( B ) Human tissue-enriched protein expression map of EXO1, PLS1, CDKN1A and FOS. ( C ) In the bone marrow, PLS1 and EXO1 are highly expressed, whereas CDKN1A and FOS are relatively low. ( D ) A represent Western blot showing overexpression of SCD1 in BM-MSCs transduced with lentivirus (“SCD1-OE”). “Control” cells are intact BM-MSCs before transduced with lentivirus. ( E ) Relative expression of mRNAs (EXO1, PLS1 FOS and CDKN1A) showing overexpression of SCD1 in BM-MSCs transduced with lentivirus (“SCD1-OE”); “Control” cells are intact BM-MSCs before transduced with lentivirus.
    Figure Legend Snippet: CDKN1A, FOS, EXO1 and PLS1 are regulated by SCD1. ( A ) Independent t -test results for the association between mRNAs (EXO1, PLS1, FOS and CDKN1A) and trauma energy in patients with clinical diabetic fractures patients. ( B ) Human tissue-enriched protein expression map of EXO1, PLS1, CDKN1A and FOS. ( C ) In the bone marrow, PLS1 and EXO1 are highly expressed, whereas CDKN1A and FOS are relatively low. ( D ) A represent Western blot showing overexpression of SCD1 in BM-MSCs transduced with lentivirus (“SCD1-OE”). “Control” cells are intact BM-MSCs before transduced with lentivirus. ( E ) Relative expression of mRNAs (EXO1, PLS1 FOS and CDKN1A) showing overexpression of SCD1 in BM-MSCs transduced with lentivirus (“SCD1-OE”); “Control” cells are intact BM-MSCs before transduced with lentivirus.

    Techniques Used: Expressing, Western Blot, Over Expression, Transduction

    30) Product Images from "Mesenchymal stem cells inhibit RANK-RANKL interactions between osteoclasts and Th17 cells via osteoprotegerin activity"

    Article Title: Mesenchymal stem cells inhibit RANK-RANKL interactions between osteoclasts and Th17 cells via osteoprotegerin activity

    Journal: Oncotarget

    doi: 10.18632/oncotarget.21379

    T-MSCs constitutively produce OPG A. The expression of surface antigens on BM-MSCs, AT-MSCs, and T-MSCs were detected by flow cytometry. Cells were negative for hematopoietic cell markers (CD14, CD34, CD45) and positive for CD73, CD90 and CD105. The data show a representative histogram from three experiments. B. BM-MSCs, AT-MSCs, and T-MSCs were harvested and the mRNA expression of TNFRSF11B (OPG encoding gene) was analyzed by real time-quantitative PCR. Data are presented as means ± SEM (* P
    Figure Legend Snippet: T-MSCs constitutively produce OPG A. The expression of surface antigens on BM-MSCs, AT-MSCs, and T-MSCs were detected by flow cytometry. Cells were negative for hematopoietic cell markers (CD14, CD34, CD45) and positive for CD73, CD90 and CD105. The data show a representative histogram from three experiments. B. BM-MSCs, AT-MSCs, and T-MSCs were harvested and the mRNA expression of TNFRSF11B (OPG encoding gene) was analyzed by real time-quantitative PCR. Data are presented as means ± SEM (* P

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction

    31) Product Images from "FOXO3 is targeted by miR-223-3p and promotes osteogenic differentiation of bone marrow mesenchymal stem cells by enhancing autophagy"

    Article Title: FOXO3 is targeted by miR-223-3p and promotes osteogenic differentiation of bone marrow mesenchymal stem cells by enhancing autophagy

    Journal: Human Cell

    doi: 10.1007/s13577-020-00421-y

    FOXO3 reversed the effects of miR-223-3p on BM-MSC osteogenic differentiation. a, b ALP levels in BM-MSCs after overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p were quantified with ELISA. c, d Effects of overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p on BM-MSC osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. e–h Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) after overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p were measured with Western blot and qRT-PCR. GAPDH was an internal control. All experiments have been performed in triplicate and experimental data were expressed as mean ± standard deviation (SD). + P
    Figure Legend Snippet: FOXO3 reversed the effects of miR-223-3p on BM-MSC osteogenic differentiation. a, b ALP levels in BM-MSCs after overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p were quantified with ELISA. c, d Effects of overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p on BM-MSC osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. e–h Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) after overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p were measured with Western blot and qRT-PCR. GAPDH was an internal control. All experiments have been performed in triplicate and experimental data were expressed as mean ± standard deviation (SD). + P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Quantitative RT-PCR, Standard Deviation

    FOXO3 expression and ALP level were increased during osteogenic differentiation. a Relative FOXO3 expressions in BM-MSCs on days 0, 3, 7, and 14 days after osteogenic differentiation were measured with quantitative real-time polymerase chain reaction (qRT-PCR). GAPDH was an internal control. b ALP levels in BM-MSCs on days 0, 3, 7, and 14 days after the osteogenic differentiation were quantified with enzyme-linked immunosorbent assay (ELISA). c BM-MSC osteogenic differentiation on days 0, 3, 7, and 14 was assessed with Alizarin Red Staining, under 100 × magnification. All the experiments have been performed in independent triplicate and experimental data were expressed as mean ± standard deviation (SD). ** P
    Figure Legend Snippet: FOXO3 expression and ALP level were increased during osteogenic differentiation. a Relative FOXO3 expressions in BM-MSCs on days 0, 3, 7, and 14 days after osteogenic differentiation were measured with quantitative real-time polymerase chain reaction (qRT-PCR). GAPDH was an internal control. b ALP levels in BM-MSCs on days 0, 3, 7, and 14 days after the osteogenic differentiation were quantified with enzyme-linked immunosorbent assay (ELISA). c BM-MSC osteogenic differentiation on days 0, 3, 7, and 14 was assessed with Alizarin Red Staining, under 100 × magnification. All the experiments have been performed in independent triplicate and experimental data were expressed as mean ± standard deviation (SD). ** P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Standard Deviation

    Effects of FOXO3 on BM-MSC osteogenic differentiation factors were detected. a, b ALP levels in BM-MSCs after up-regulating or down-regulating FOXO3 were quantified with ELISA. c Effects of up-regulating or down-regulating FOXO3 on BM-MSC osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. d–g Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) were measured with Western blot and qRT-PCR. GAPDH was an internal control. All the experiments have been performed in independent triplicate and the experimental data were expressed as mean ± standard deviation (SD). ++ P
    Figure Legend Snippet: Effects of FOXO3 on BM-MSC osteogenic differentiation factors were detected. a, b ALP levels in BM-MSCs after up-regulating or down-regulating FOXO3 were quantified with ELISA. c Effects of up-regulating or down-regulating FOXO3 on BM-MSC osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. d–g Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) were measured with Western blot and qRT-PCR. GAPDH was an internal control. All the experiments have been performed in independent triplicate and the experimental data were expressed as mean ± standard deviation (SD). ++ P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Quantitative RT-PCR, Standard Deviation

    Effects of FOXO3 on BM-MSCs osteogenic differentiation via enhancing autophagy-related genes expressions were detected. a, b ALP levels in BM-MSCs after overexpressed FOXO3 and 3-MA treatment or siFOXO3 and RAPA treatment were quantified with ELISA. c, d Effects of overexpressed FOXO3 and 3-MA treatment or siFOXO3 and RAPA treatment on BM-MSCs osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. e, h Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) after overexpressing FOXO3 or siFOXO3 and 3-MA or RAPA treatment were measured with Western blot and qRT-PCR. GAPDH was an internal control. All experiments have been performed in triplicate and experimental data were expressed as mean ± standard deviation (SD). ++ P
    Figure Legend Snippet: Effects of FOXO3 on BM-MSCs osteogenic differentiation via enhancing autophagy-related genes expressions were detected. a, b ALP levels in BM-MSCs after overexpressed FOXO3 and 3-MA treatment or siFOXO3 and RAPA treatment were quantified with ELISA. c, d Effects of overexpressed FOXO3 and 3-MA treatment or siFOXO3 and RAPA treatment on BM-MSCs osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. e, h Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) after overexpressing FOXO3 or siFOXO3 and 3-MA or RAPA treatment were measured with Western blot and qRT-PCR. GAPDH was an internal control. All experiments have been performed in triplicate and experimental data were expressed as mean ± standard deviation (SD). ++ P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Quantitative RT-PCR, Standard Deviation

    32) Product Images from "LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK"

    Article Title: LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK

    Journal: Stem Cells (Dayton, Ohio)

    doi: 10.1002/stem.2937

    Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p
    Figure Legend Snippet: Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p

    Techniques Used: Expressing, Over Expression, Derivative Assay, Western Blot, Staining, Activity Assay

    33) Product Images from "Potency testing of mesenchymal stromal cell growth expanded in human platelet lysate from different human tissues"

    Article Title: Potency testing of mesenchymal stromal cell growth expanded in human platelet lysate from different human tissues

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-016-0383-3

    Immune-related markers. a Histograms showing the immune-related antigen expression of BM-MSCs, UCT-MSCs and AT-MSCs. One representative MSC sample for each source is shown. b Quantitative expression of immune-related markers measured by flow cytometry. Results are expressed as mean ± SD (standard deviation) of five samples processed for each tissue. AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue
    Figure Legend Snippet: Immune-related markers. a Histograms showing the immune-related antigen expression of BM-MSCs, UCT-MSCs and AT-MSCs. One representative MSC sample for each source is shown. b Quantitative expression of immune-related markers measured by flow cytometry. Results are expressed as mean ± SD (standard deviation) of five samples processed for each tissue. AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Standard Deviation

    CFSE dilution profile in CFSE-based lymphocyte proliferation assay. Histograms representing the distribution of proliferating lymphocytes with respect to CFSE expression level. When labelled cells undergo cell division, the CFSE fluorescence intensity is reduced by 50 %, generating new peaks on the left side of the initial peak of dye intensity. CFSE dilution profiles of total proliferating lymphocytes at 72 h, 96 h and 120 h in the absence (CTRL) or in the presence of BM-MSCs, UCT-MSCs and AT-MSCs are shown. AT adipose tissue, BM bone marrow, CTRL control, MSCs mesenchymal stromal cells, UCT umbilical cord tissue
    Figure Legend Snippet: CFSE dilution profile in CFSE-based lymphocyte proliferation assay. Histograms representing the distribution of proliferating lymphocytes with respect to CFSE expression level. When labelled cells undergo cell division, the CFSE fluorescence intensity is reduced by 50 %, generating new peaks on the left side of the initial peak of dye intensity. CFSE dilution profiles of total proliferating lymphocytes at 72 h, 96 h and 120 h in the absence (CTRL) or in the presence of BM-MSCs, UCT-MSCs and AT-MSCs are shown. AT adipose tissue, BM bone marrow, CTRL control, MSCs mesenchymal stromal cells, UCT umbilical cord tissue

    Techniques Used: Lymphocyte Proliferation Assay, Expressing, Fluorescence

    MSC immunosuppressive effect on lymphocyte proliferation. Data show the percentages of residual proliferative capacity of stimulated lymphocytes in the absence (CTRL) or in the presence of BM-MSCs, UCT-MSCs and AT-MSCs at ratio 1:4 and 1:8 (MSCs:PBMCs), either in cell-cell contact or in transwell system, evaluated at 72 h, 96 h and 120 h. (F = 29.025 and p = 0.000 at ANOVA and * p = 0.000 for BM vs AT, * p = 0.001 for BM vs UCT, # p = 0.029 for AT vs UCT at Scheffe test for 1:4 ratio in contact cultures; F = 53.301 and p = 0.000 at ANOVA and ° p = 0.000 for BM vs AT, ° p = 0.001 for BM vs UCT, p = 0.237 for AT vs UCT at Scheffe test for 1:4 ratio for transwell cultures; F = 6.57 and p = 0.008 at ANOVA and p = 0.152 for BM vs AT, § p = 0.009 for BM vs UCT, p = 0.328 for AT vs UCT at Scheffe test for 1:8 ratio in contact cultures; F = 2.86 and p = 0.088 at ANOVA and p = 0.151 for BM vs AT, p = 0.152 for BM vs UCT, p = 0.981 for AT vs UCT at Scheffe test for 1:8 ratio in transwell cultures). AT adipose tissue, BM bone marrow, CTRL control, UCT umbilical cord tissue
    Figure Legend Snippet: MSC immunosuppressive effect on lymphocyte proliferation. Data show the percentages of residual proliferative capacity of stimulated lymphocytes in the absence (CTRL) or in the presence of BM-MSCs, UCT-MSCs and AT-MSCs at ratio 1:4 and 1:8 (MSCs:PBMCs), either in cell-cell contact or in transwell system, evaluated at 72 h, 96 h and 120 h. (F = 29.025 and p = 0.000 at ANOVA and * p = 0.000 for BM vs AT, * p = 0.001 for BM vs UCT, # p = 0.029 for AT vs UCT at Scheffe test for 1:4 ratio in contact cultures; F = 53.301 and p = 0.000 at ANOVA and ° p = 0.000 for BM vs AT, ° p = 0.001 for BM vs UCT, p = 0.237 for AT vs UCT at Scheffe test for 1:4 ratio for transwell cultures; F = 6.57 and p = 0.008 at ANOVA and p = 0.152 for BM vs AT, § p = 0.009 for BM vs UCT, p = 0.328 for AT vs UCT at Scheffe test for 1:8 ratio in contact cultures; F = 2.86 and p = 0.088 at ANOVA and p = 0.151 for BM vs AT, p = 0.152 for BM vs UCT, p = 0.981 for AT vs UCT at Scheffe test for 1:8 ratio in transwell cultures). AT adipose tissue, BM bone marrow, CTRL control, UCT umbilical cord tissue

    Techniques Used:

    Platelet lysate lot testing. a MSCs growth assay with different platelet lysate lots. Two MSCs cell lines for each source (BM, UCT, AT) were used to test the growth promotion ability and variability of eight different PL lots (PL1–PL8). MSCs proliferation was evaluated at passage 1 (P1) and passage 2 (P2). b Coefficient of variation percentage related to the eight different PL lots (PL1–PL8) in BM-MSCs, UCT-MSCs, and AT-MSCs cultures at P1 and P2. Results are expressed as mean ± SD (standard deviation). (F = 0.479 and p = 0.847 at ANOVA and p > 0.900 for any post hoc comparison at Scheffe test) AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue
    Figure Legend Snippet: Platelet lysate lot testing. a MSCs growth assay with different platelet lysate lots. Two MSCs cell lines for each source (BM, UCT, AT) were used to test the growth promotion ability and variability of eight different PL lots (PL1–PL8). MSCs proliferation was evaluated at passage 1 (P1) and passage 2 (P2). b Coefficient of variation percentage related to the eight different PL lots (PL1–PL8) in BM-MSCs, UCT-MSCs, and AT-MSCs cultures at P1 and P2. Results are expressed as mean ± SD (standard deviation). (F = 0.479 and p = 0.847 at ANOVA and p > 0.900 for any post hoc comparison at Scheffe test) AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue

    Techniques Used: Growth Assay, Standard Deviation

    Expansion potential of MSCs from BM, UCT and AT. a The expansion of MSCs from ten samples of each source (BM, UCT, AT) revealed a high inter-individual variability in the amount of MSCs obtained per gram of initial tissue, which ranged between 2 × 10 6 and 80 × 10 6 at Passage 2 (P2). b Comparison of the average of MSCs cumulative cell yield per gram of initial tissue (BM, UCT, AT) at Passage 1 (P1) and P2. Results were represented as mean ± SEM (standard error of the mean) (F = 19.712 and p
    Figure Legend Snippet: Expansion potential of MSCs from BM, UCT and AT. a The expansion of MSCs from ten samples of each source (BM, UCT, AT) revealed a high inter-individual variability in the amount of MSCs obtained per gram of initial tissue, which ranged between 2 × 10 6 and 80 × 10 6 at Passage 2 (P2). b Comparison of the average of MSCs cumulative cell yield per gram of initial tissue (BM, UCT, AT) at Passage 1 (P1) and P2. Results were represented as mean ± SEM (standard error of the mean) (F = 19.712 and p

    Techniques Used:

    Flow cytometric analysis of MSCs phenotype. a Histograms showing the MSCs, hematopoietic and endothelial surface antigen expression of BM-MSCs, UCT-MSCs and AT-MSCs. One representative MSC sample for each source is shown. b Quantitative expression of MSCs, hematopoietic and endothelial antigens measured by flow cytometry. Results are expressed as mean ± SD (standard deviation) of ten samples processed for each tissue. AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue
    Figure Legend Snippet: Flow cytometric analysis of MSCs phenotype. a Histograms showing the MSCs, hematopoietic and endothelial surface antigen expression of BM-MSCs, UCT-MSCs and AT-MSCs. One representative MSC sample for each source is shown. b Quantitative expression of MSCs, hematopoietic and endothelial antigens measured by flow cytometry. Results are expressed as mean ± SD (standard deviation) of ten samples processed for each tissue. AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue

    Techniques Used: Flow Cytometry, Expressing, Cytometry, Standard Deviation

    Morphology and differentiation assays of MSCs. Spindle-shaped adherent cells with MSCs morphology were observed in BM-MSCs ( a ), UCT-MSCs ( b ) and AT-MSCs ( c ) cultures. Representative images of BM-MSCs, UCT-MSCs and AT-MSCs induced to differentiate into adipogenic ( d - f ), osteogenic ( g - i ), and chondrogenic lineages ( l - n ). (Magnification × 100). AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue
    Figure Legend Snippet: Morphology and differentiation assays of MSCs. Spindle-shaped adherent cells with MSCs morphology were observed in BM-MSCs ( a ), UCT-MSCs ( b ) and AT-MSCs ( c ) cultures. Representative images of BM-MSCs, UCT-MSCs and AT-MSCs induced to differentiate into adipogenic ( d - f ), osteogenic ( g - i ), and chondrogenic lineages ( l - n ). (Magnification × 100). AT adipose tissue, BM bone marrow, MSCs mesenchymal stromal cells, UCT umbilical cord tissue

    Techniques Used:

    34) Product Images from "Bone marrow-derived humoral factors suppress oxidative phosphorylation, upregulate TSG-6, and improve therapeutic effects on liver injury of mesenchymal stem cells"

    Article Title: Bone marrow-derived humoral factors suppress oxidative phosphorylation, upregulate TSG-6, and improve therapeutic effects on liver injury of mesenchymal stem cells

    Journal: Journal of Clinical Biochemistry and Nutrition

    doi: 10.3164/jcbn.19-125

    Effects of removing particles over 20 nm from BMC-CM. Using the ExoMir TM PLUS kit, particles over 20 nm were removed from BMC-CM (filtered BMC-CM), and exosomal microRNAs contained in BMC-CM were extracted (A). Mitochondrial OXPHOS activity was accelerated, and the spare respiratory capacity was significantly increased in filtered BMC-CM compared with non-filtered BMC-CM in cells at passage 1 (B). Expression of Hif1a tended to decrease, and that of Sirt3 , Sirt4 , and Sirt5 were significantly reduced by filtered BMC-CM in cells at passage 3 (C). Pathway analysis of mRNA-microarray data from BM-MSCs cultured in BMC-CM vs filtered BMC-CM analyzed by IPA revealed that 20–220-nm particles were involved in multiple signaling pathways such as OXPHOS, Sirtuin signaling, mTOR signaling, PI3K/Akt signaling, and IGF-1 signaling pathways (D). The overlap p value ( p value), indicator of overlap between observed gene expression changes and known targets regulated by transcriptional regulators, was calculated using Fisher’s exact test. The activated z-score (z-score), indicator of regulation direction of pathways, was calculated based on the database of molecular network that represents experimentally observed gene expression or transcription events; positive z-score means “activating” and negative “inhibiting”. OXPHOS-pathway map by pathway analysis of IPA showed that all of the mitochondrial complexes I to V were suppressed by 20–220-nm particles in BMC-CM. Up-regulated components are colored red, and down-regulated green (E). Error bars indicate SE. * p
    Figure Legend Snippet: Effects of removing particles over 20 nm from BMC-CM. Using the ExoMir TM PLUS kit, particles over 20 nm were removed from BMC-CM (filtered BMC-CM), and exosomal microRNAs contained in BMC-CM were extracted (A). Mitochondrial OXPHOS activity was accelerated, and the spare respiratory capacity was significantly increased in filtered BMC-CM compared with non-filtered BMC-CM in cells at passage 1 (B). Expression of Hif1a tended to decrease, and that of Sirt3 , Sirt4 , and Sirt5 were significantly reduced by filtered BMC-CM in cells at passage 3 (C). Pathway analysis of mRNA-microarray data from BM-MSCs cultured in BMC-CM vs filtered BMC-CM analyzed by IPA revealed that 20–220-nm particles were involved in multiple signaling pathways such as OXPHOS, Sirtuin signaling, mTOR signaling, PI3K/Akt signaling, and IGF-1 signaling pathways (D). The overlap p value ( p value), indicator of overlap between observed gene expression changes and known targets regulated by transcriptional regulators, was calculated using Fisher’s exact test. The activated z-score (z-score), indicator of regulation direction of pathways, was calculated based on the database of molecular network that represents experimentally observed gene expression or transcription events; positive z-score means “activating” and negative “inhibiting”. OXPHOS-pathway map by pathway analysis of IPA showed that all of the mitochondrial complexes I to V were suppressed by 20–220-nm particles in BMC-CM. Up-regulated components are colored red, and down-regulated green (E). Error bars indicate SE. * p

    Techniques Used: Activity Assay, Expressing, Microarray, Cell Culture, Indirect Immunoperoxidase Assay

    Whole bone marrow cell-derived miRNAs may improve the therapeutic quality of MSCs. Of the seven miRNAs used for transfection experiments, miR-23b-3p, miR-92b-3p, miR-204b-3p, miR-326b-5p, and miR-1247b-3p significantly upregulated one or more mRNA ( Hif1a , Sirt3 , Sirt4 , Sirt5 , and Tsg6 ) Error bars indicate SE. * p
    Figure Legend Snippet: Whole bone marrow cell-derived miRNAs may improve the therapeutic quality of MSCs. Of the seven miRNAs used for transfection experiments, miR-23b-3p, miR-92b-3p, miR-204b-3p, miR-326b-5p, and miR-1247b-3p significantly upregulated one or more mRNA ( Hif1a , Sirt3 , Sirt4 , Sirt5 , and Tsg6 ) Error bars indicate SE. * p

    Techniques Used: Derivative Assay, Transfection

    35) Product Images from "Haemin pre‐treatment augments the cardiac protection of mesenchymal stem cells by inhibiting mitochondrial fission and improving survival, et al. Haemin pre‐treatment augments the cardiac protection of mesenchymal stem cells by inhibiting mitochondrial fission and improving survival"

    Article Title: Haemin pre‐treatment augments the cardiac protection of mesenchymal stem cells by inhibiting mitochondrial fission and improving survival, et al. Haemin pre‐treatment augments the cardiac protection of mesenchymal stem cells by inhibiting mitochondrial fission and improving survival

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14747

    Haemin pre‐treatment inhibits mitochondrial fragmentation and apoptosis of bone marrow‐mesenchymal stem cell (BM‐MSCs) by regulating HO‐1. A, Western blotting and quantitative analysis for the expression of HO‐1 in BM‐MSCs or haemin‐pretreated BM‐MSCs under normoxia or SD/H. B, Western blotting and quantitative analysis for the expression of HO‐1 in BM‐MSCs and haemin‐pretreated BM‐MSCs treated with control siRNA or HO‐1 siRNA under SD/H. C, Representative images of the fragmented mitochondria (magnification of 20x) and quantitative analysis of fragmented mitochondria in BM‐MSCs and haemin‐pretreated BM‐MSCs treated with control siRNA or HO‐1 siRNA under SD/H. D, Representative images of TUNEL staining (magnification of 20x) and quantitative analysis of the apoptosis of BM‐MSCs and haemin‐pretreated BM‐MSCs treated with control siRNA or HO‐1 siRNA under SD/H. Data are expressed as the mean ± SEM. n = 3. Scale bar = 50 μm * P
    Figure Legend Snippet: Haemin pre‐treatment inhibits mitochondrial fragmentation and apoptosis of bone marrow‐mesenchymal stem cell (BM‐MSCs) by regulating HO‐1. A, Western blotting and quantitative analysis for the expression of HO‐1 in BM‐MSCs or haemin‐pretreated BM‐MSCs under normoxia or SD/H. B, Western blotting and quantitative analysis for the expression of HO‐1 in BM‐MSCs and haemin‐pretreated BM‐MSCs treated with control siRNA or HO‐1 siRNA under SD/H. C, Representative images of the fragmented mitochondria (magnification of 20x) and quantitative analysis of fragmented mitochondria in BM‐MSCs and haemin‐pretreated BM‐MSCs treated with control siRNA or HO‐1 siRNA under SD/H. D, Representative images of TUNEL staining (magnification of 20x) and quantitative analysis of the apoptosis of BM‐MSCs and haemin‐pretreated BM‐MSCs treated with control siRNA or HO‐1 siRNA under SD/H. Data are expressed as the mean ± SEM. n = 3. Scale bar = 50 μm * P

    Techniques Used: Western Blot, Expressing, TUNEL Assay, Staining

    36) Product Images from "Arrhythmogenic Cardiomyopathy Is a Multicellular Disease Affecting Cardiac and Bone Marrow Mesenchymal Stromal Cells"

    Article Title: Arrhythmogenic Cardiomyopathy Is a Multicellular Disease Affecting Cardiac and Bone Marrow Mesenchymal Stromal Cells

    Journal: Journal of Clinical Medicine

    doi: 10.3390/jcm10091871

    AC-linked Dsg2 variant affects cytoskeletal organization and morphology of cardiac and bone marrow mesenchymal stromal cells. ( a – d ) Confocal IF analysis of cultured cardiac- and BM-MSCs isolated from control or Dsg2 mut/mut mice. Cells were stained with AlexaFLUOR ® -568 conjugated phalloidin (red signal). Nuclei were counterstained with DAPI (blue signal). White arrows indicate focal adhesions, and arrowheads evidence actin puncta. Insets show sub-membrane thick actin filaments. ( e – g ) Morphometric evaluation of cell surface, perimeter, and perimeter/area ratio in cultured cardiac and BM-MSCs isolated from control or Dsg2 mut/mut mice. A total of 110 cells for each study group were analyzed. Whiskers represent min to max values. **** p ≤ 0.0001.
    Figure Legend Snippet: AC-linked Dsg2 variant affects cytoskeletal organization and morphology of cardiac and bone marrow mesenchymal stromal cells. ( a – d ) Confocal IF analysis of cultured cardiac- and BM-MSCs isolated from control or Dsg2 mut/mut mice. Cells were stained with AlexaFLUOR ® -568 conjugated phalloidin (red signal). Nuclei were counterstained with DAPI (blue signal). White arrows indicate focal adhesions, and arrowheads evidence actin puncta. Insets show sub-membrane thick actin filaments. ( e – g ) Morphometric evaluation of cell surface, perimeter, and perimeter/area ratio in cultured cardiac and BM-MSCs isolated from control or Dsg2 mut/mut mice. A total of 110 cells for each study group were analyzed. Whiskers represent min to max values. **** p ≤ 0.0001.

    Techniques Used: Variant Assay, Cell Culture, Isolation, Mouse Assay, Staining

    DSG2-downregulated bone marrow mesenchymal stromal cells display altered cytoskeletal organization. ( a ) Western blotting on protein extracts from cultured rat BM-MSCs transduced with either Ad-Empty or Ad-sh Dsg2 at different MOI (25, 50, and 100). GAPDH was used to ensure equal protein loading. MW, molecular weight. C-, negative control ( Dsg2 mut/mut heart). ( b ) Confocal IF analysis of cultured rat BM-MSCs infected with Ad-Empty (MOI 50) or Ad-sh Dsg2 (MOI 50). Cells were stained with AlexaFLUOR ® -568 conjugated phalloidin (red signal). Nuclei were counterstained with DAPI (blue signal). ( c – e ) Morphometric evaluation of cell surface, perimeter, and perimeter/area ratio in cultured rat BM-MSCs shown in ( b ). A total of 25 cells for each study group were analyzed. Whiskers represent min to max values. ** p ≤ 0.01; **** p ≤ 0.0001. ( f ) Confocal IF analysis of cultured rat BM-MSCs infected with Ad-Empty (MOI 50) and Ad-sh Dsg2 (MOI 50). Cells were stained with anti-α-tubulin antibody (green signal). Nuclei were counterstained with DAPI (blue signal).
    Figure Legend Snippet: DSG2-downregulated bone marrow mesenchymal stromal cells display altered cytoskeletal organization. ( a ) Western blotting on protein extracts from cultured rat BM-MSCs transduced with either Ad-Empty or Ad-sh Dsg2 at different MOI (25, 50, and 100). GAPDH was used to ensure equal protein loading. MW, molecular weight. C-, negative control ( Dsg2 mut/mut heart). ( b ) Confocal IF analysis of cultured rat BM-MSCs infected with Ad-Empty (MOI 50) or Ad-sh Dsg2 (MOI 50). Cells were stained with AlexaFLUOR ® -568 conjugated phalloidin (red signal). Nuclei were counterstained with DAPI (blue signal). ( c – e ) Morphometric evaluation of cell surface, perimeter, and perimeter/area ratio in cultured rat BM-MSCs shown in ( b ). A total of 25 cells for each study group were analyzed. Whiskers represent min to max values. ** p ≤ 0.01; **** p ≤ 0.0001. ( f ) Confocal IF analysis of cultured rat BM-MSCs infected with Ad-Empty (MOI 50) and Ad-sh Dsg2 (MOI 50). Cells were stained with anti-α-tubulin antibody (green signal). Nuclei were counterstained with DAPI (blue signal).

    Techniques Used: Western Blot, Cell Culture, Transduction, Molecular Weight, Negative Control, Infection, Staining

    37) Product Images from "Mitochondria transfer from mesenchymal stem cells structurally and functionally repairs renal proximal tubular epithelial cells in diabetic nephropathy in vivo"

    Article Title: Mitochondria transfer from mesenchymal stem cells structurally and functionally repairs renal proximal tubular epithelial cells in diabetic nephropathy in vivo

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-40163-y

    Incorporation of BM-MSC-Mt into STZ-PTECs, and the anti-degenerative effects of BM-MSCs and BM-MSC-Mt in vitro . ( a ) Time-lapse images of Mt transfer from MtDsRed2-MSCs to STZ-PTECs. Images were obtained 4, 5 and 6 h after commencing time-lapse observations. Panels on the right show magnified images of the left panels. White arrows track the same DsRed2-Mt. M: MtDsRed2-MSCs, P: PTECs derived from STZ rats (STZ-PTECs). Scale bar, 25 µm. ( b ) Time-lapse images of the incorporation of isolated Mt (BM-MSC-Mt) into STZ-PTECs. Images were obtained at 10 and 15 min and 4 h after commencing time-lapse observations. Panels on the right show magnified images of left panels. White arrows and arrowheads track the same DsRed2-Mt. P: PTECs derived from STZ rats (STZ-PTECs). Scale bar, 25 µm. ( c ) Phase contrast observations of Control-PTECs and STZ-PTECs cultured with or without BM-MSCs and BM-MSC-Mt. Images were obtained 12, 48 and 96 h after commencing the co-culture with BM-MSCs or BM-MSC-Mt. Scale bar, 100 µm.
    Figure Legend Snippet: Incorporation of BM-MSC-Mt into STZ-PTECs, and the anti-degenerative effects of BM-MSCs and BM-MSC-Mt in vitro . ( a ) Time-lapse images of Mt transfer from MtDsRed2-MSCs to STZ-PTECs. Images were obtained 4, 5 and 6 h after commencing time-lapse observations. Panels on the right show magnified images of the left panels. White arrows track the same DsRed2-Mt. M: MtDsRed2-MSCs, P: PTECs derived from STZ rats (STZ-PTECs). Scale bar, 25 µm. ( b ) Time-lapse images of the incorporation of isolated Mt (BM-MSC-Mt) into STZ-PTECs. Images were obtained at 10 and 15 min and 4 h after commencing time-lapse observations. Panels on the right show magnified images of left panels. White arrows and arrowheads track the same DsRed2-Mt. P: PTECs derived from STZ rats (STZ-PTECs). Scale bar, 25 µm. ( c ) Phase contrast observations of Control-PTECs and STZ-PTECs cultured with or without BM-MSCs and BM-MSC-Mt. Images were obtained 12, 48 and 96 h after commencing the co-culture with BM-MSCs or BM-MSC-Mt. Scale bar, 100 µm.

    Techniques Used: In Vitro, Derivative Assay, Isolation, Cell Culture, Co-Culture Assay

    Localisation of BM-MSC-Mt injected under the renal capsule and histological improvement effects in STZ rats. ( a ) Experimental protocol for the injection of BM-MSC-Mt in STZ rats. Isolated DsRed2-Mt obtained from 1 × 10 6 MtDsRed2-MSCs were injected under the renal capsule on the left side of the kidney. An equal volume of PBS was administered to the right kidney as the vehicle ( n = 3 per group). ( b ) Immunofluorescence images of megalin (green) expression and localisation of isolated DsRed2-Mt (red) in the kidney of STZ rats. Nuclei are counterstained with DAPI (blue). PT, proximal tubules. Scale bar, 20 µm. ( c ) Light microscopic images of proximal tubules in STZ rats. Kidney sections were stained with H E. Black arrowheads indicate injected isolated DsRed2-Mt stained by nickel-enhanced DAB. PT, proximal tubules. Scale bar, 20 µm in left and middle panels; 10 nm in inset. ( d ) Immunofluorescence images of collagen IV (green) expression in proximal tubules of STZ rats. Nuclei are counterstained with DAPI (blue). White arrows indicate an injured tubular basement membrane with loss or weakened expression of collagen IV. White arrowheads indicate degenerated nuclei. PT, proximal tubules. Scale bar, 20 µm. ( e ) Immunofluorescence images of megalin (green) expression in proximal tubules of STZ rats. Nuclei are counterstained with DAPI (blue). White arrows show degenerated nuclei. PT, proximal tubules. Scale bar, 20 µm.
    Figure Legend Snippet: Localisation of BM-MSC-Mt injected under the renal capsule and histological improvement effects in STZ rats. ( a ) Experimental protocol for the injection of BM-MSC-Mt in STZ rats. Isolated DsRed2-Mt obtained from 1 × 10 6 MtDsRed2-MSCs were injected under the renal capsule on the left side of the kidney. An equal volume of PBS was administered to the right kidney as the vehicle ( n = 3 per group). ( b ) Immunofluorescence images of megalin (green) expression and localisation of isolated DsRed2-Mt (red) in the kidney of STZ rats. Nuclei are counterstained with DAPI (blue). PT, proximal tubules. Scale bar, 20 µm. ( c ) Light microscopic images of proximal tubules in STZ rats. Kidney sections were stained with H E. Black arrowheads indicate injected isolated DsRed2-Mt stained by nickel-enhanced DAB. PT, proximal tubules. Scale bar, 20 µm in left and middle panels; 10 nm in inset. ( d ) Immunofluorescence images of collagen IV (green) expression in proximal tubules of STZ rats. Nuclei are counterstained with DAPI (blue). White arrows indicate an injured tubular basement membrane with loss or weakened expression of collagen IV. White arrowheads indicate degenerated nuclei. PT, proximal tubules. Scale bar, 20 µm. ( e ) Immunofluorescence images of megalin (green) expression in proximal tubules of STZ rats. Nuclei are counterstained with DAPI (blue). White arrows show degenerated nuclei. PT, proximal tubules. Scale bar, 20 µm.

    Techniques Used: Injection, Isolation, Immunofluorescence, Expressing, Staining

    38) Product Images from "LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK"

    Article Title: LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK

    Journal: Stem Cells (Dayton, Ohio)

    doi: 10.1002/stem.2937

    Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p
    Figure Legend Snippet: Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p

    Techniques Used: Expressing, Over Expression, Derivative Assay, Western Blot, Staining, Activity Assay

    39) Product Images from "LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK"

    Article Title: LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK

    Journal: Stem Cells (Dayton, Ohio)

    doi: 10.1002/stem.2937

    Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p
    Figure Legend Snippet: Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p

    Techniques Used: Expressing, Over Expression, Derivative Assay, Western Blot, Staining, Activity Assay

    40) Product Images from "LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK"

    Article Title: LncRNA‐OG Promotes the Osteogenic Differentiation of Bone Marrow‐Derived Mesenchymal Stem Cells Under the Regulation of hnRNPK

    Journal: Stem Cells (Dayton, Ohio)

    doi: 10.1002/stem.2937

    Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p
    Figure Legend Snippet: Osteogenesis‐associated long noncoding RNA (lncRNA‐OG) regulates the expression of bone morphogenetic protein (BMP) family proteins by interacting with hnRNPK. (A): Expression analysis of BMP signaling, β‐catenin/WNT signaling and ERK1/2/MAPK signaling following lncRNA‐OG knockdown and overexpression in bone marrow‐derived mesenchymal stem cells (BM‐MSCs) by Western blotting. GAPDH was used as the internal control (left panel). Quantification of pSmad1/5/8 band intensities (right panel). (B): Related mRNA levels of BMP2 , BMP4 , BMP6 , BMP7 , and BMP9 after lncRNA‐OG knockdown on day 7 of osteogenic differentiation. Data were normalized to GAPDH. (C): Expression of BMP family proteins following lncRNA‐OG and hnRNPK knockdown respectively in BM‐MSCs by Western blotting. GAPDH was used as the internal control. (D): Left: alkaline phosphatase (ALP) staining on day 7 (upper panel), alizarin red (ARS) staining on day 14 after osteogenic induction (lower panel). Right: ALP activity was determined as units per gram of protein per 15 minutes. ARS staining was quantified as the absorbance at 562 nm. Data are presented as the mean ± SD. *, p

    Techniques Used: Expressing, Over Expression, Derivative Assay, Western Blot, Staining, Activity Assay

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    Flow Cytometry:

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    Cytometry:

    Article Title: Expression of hsa-MIR-204, RUNX2, PPARγ, and BCL2 in Bone Marrow Derived Mesenchymal Stem Cells from Multiple Myeloma Patients and Normal Individuals
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    Labeling:

    Article Title: Expression of hsa-MIR-204, RUNX2, PPARγ, and BCL2 in Bone Marrow Derived Mesenchymal Stem Cells from Multiple Myeloma Patients and Normal Individuals
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    other:

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    Incubation:

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    Expressing:

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    Microarray:

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    In Vivo:

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    Migration:

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    Fluorescence:

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    Imaging:

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    Article Snippet: .. For in vivo migration studies, BM-MSCs or BM-MSCs pretreated with rAMF (5×105 ) were prelabeled with CMDiI for histological analysis and DiR (Molecular Probes, Invitrogen) for fluorescence imaging (FI). .. FI was performed using the Xenogen In Vivo Imaging System (IVIS; Caliper Life Sciences, Hopkinton, MA, USA) 1 hour after MSC injection and every day until experimental end point.

    Staining:

    Article Title: Engineered Stem Cells Improve Neurogenic Bladder by Overexpressing SDF-1 in a Pelvic Nerve Injury Rat Model
    Article Snippet: The supernatant collected from BM-MSCs, imMSCs/eSDF-1− , and imMSCs/eSDF-1+ conditioned media (CM) was used to examine the SDF-1 in each group by ELISA (R & D Systems Europe, Abingdon, UK) according to the manufacturer’s protocol. .. Meanwhile for comparison, we stained BM-MSCs, imMSCs/eSDF-1− , and imMSCs/eSDF-1+ by Cell Tracker™ CM-DiI (Molecular Probes, Eugene, OR, USA). .. Then the same ELISA process proceeded in staining group.

    Isolation:

    Article Title: Sdf-1 (CXCL12) induces CD9 expression in stem cells engaged in muscle regeneration
    Article Snippet: .. Analyses of mRNA levels in BM-MSCs and ESCs included RNA isolation using the mirVana kit (Life Technologies) and then reverse transcription using Superscript (Life Technologies). .. The TaqMan assays (Life Technologies) and Master Mix (Life Technologies) were used to analyse the level of the genes CXCR4, CXCR7, and CD9 according to the PCR array manufacturer’s instructions.

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    Thermo Fisher cells mscs
    Gene expression analysis for fold-change of chondrogenic markers in physioxia relative to hyperoxia demonstrates that high-GAG groups of both <t>MSCs</t> and <t>ACPs</t> are highly responsive to oxygen level and upregulate a majority of genes representative of the articular cartilage phenotype in low-oxygen environments. Data are mean ± standard deviation of fold-change in gene expression for each group ( n = 8). Statistical significance defined as * p
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    Gene expression analysis for fold-change of chondrogenic markers in physioxia relative to hyperoxia demonstrates that high-GAG groups of both MSCs and ACPs are highly responsive to oxygen level and upregulate a majority of genes representative of the articular cartilage phenotype in low-oxygen environments. Data are mean ± standard deviation of fold-change in gene expression for each group ( n = 8). Statistical significance defined as * p

    Journal: Stem Cell Research & Therapy

    Article Title: Responses to altered oxygen tension are distinct between human stem cells of high and low chondrogenic capacity

    doi: 10.1186/s13287-016-0419-8

    Figure Lengend Snippet: Gene expression analysis for fold-change of chondrogenic markers in physioxia relative to hyperoxia demonstrates that high-GAG groups of both MSCs and ACPs are highly responsive to oxygen level and upregulate a majority of genes representative of the articular cartilage phenotype in low-oxygen environments. Data are mean ± standard deviation of fold-change in gene expression for each group ( n = 8). Statistical significance defined as * p

    Article Snippet: Pellet cultures were formed by centrifuging 5 × 104 cells (MSCs) or 1 × 105 cells (ACPs) at 500 × g for 5 min in 240 μl of medium in Nunc polypropylene V-bottom 96-well plates (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Expressing, Standard Deviation

    a Representative toluidine blue stain for total proteoglycans demonstrates smaller pellets with less metachromasia for low-GAG MSC preparations and ACP clones in both hyperoxia and physioxia relative to paired high-GAG MSC preparations and ACP clones at the respective oxygen levels. Images were acquired with bright-field microscopy, scale bars = 100 μm. b Measurement of pellet diameter revealed a statistically significant difference in pellet size between both MSCs and ACPs of high or low chondrogenicity and between high-GAG MSCs at physioxia or hyperoxia. Statistical significance defined as * p

    Journal: Stem Cell Research & Therapy

    Article Title: Responses to altered oxygen tension are distinct between human stem cells of high and low chondrogenic capacity

    doi: 10.1186/s13287-016-0419-8

    Figure Lengend Snippet: a Representative toluidine blue stain for total proteoglycans demonstrates smaller pellets with less metachromasia for low-GAG MSC preparations and ACP clones in both hyperoxia and physioxia relative to paired high-GAG MSC preparations and ACP clones at the respective oxygen levels. Images were acquired with bright-field microscopy, scale bars = 100 μm. b Measurement of pellet diameter revealed a statistically significant difference in pellet size between both MSCs and ACPs of high or low chondrogenicity and between high-GAG MSCs at physioxia or hyperoxia. Statistical significance defined as * p

    Article Snippet: Pellet cultures were formed by centrifuging 5 × 104 cells (MSCs) or 1 × 105 cells (ACPs) at 500 × g for 5 min in 240 μl of medium in Nunc polypropylene V-bottom 96-well plates (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Staining, Clone Assay, Microscopy

    Clustering of oxygen-dependent gene expression based on z -score demonstrates that groups of MSCs and ACPs are more similar between GAG level than within cell type in response to culture in physioxia relative to culture in hyperoxia. ACP articular cartilage progenitor, GAG glycosaminoglycan, MSC mesenchymal stem cell

    Journal: Stem Cell Research & Therapy

    Article Title: Responses to altered oxygen tension are distinct between human stem cells of high and low chondrogenic capacity

    doi: 10.1186/s13287-016-0419-8

    Figure Lengend Snippet: Clustering of oxygen-dependent gene expression based on z -score demonstrates that groups of MSCs and ACPs are more similar between GAG level than within cell type in response to culture in physioxia relative to culture in hyperoxia. ACP articular cartilage progenitor, GAG glycosaminoglycan, MSC mesenchymal stem cell

    Article Snippet: Pellet cultures were formed by centrifuging 5 × 104 cells (MSCs) or 1 × 105 cells (ACPs) at 500 × g for 5 min in 240 μl of medium in Nunc polypropylene V-bottom 96-well plates (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Expressing

    Gene expression analysis for fold-change of chondrogenic markers of the articular cartilage phenotype ( COL2A1 , ACAN ), the fibrocartilaginous phenotype ( COL1A1 ), and the hypertrophic phenotype ( COL10A1 , MMP13 ) demonstrates varied chondrogenic responses by high-GAG and low-GAG groups of each cell type, MSCs and ACPs, during pellet culture in physioxic relative to hyperoxic conditions. Data are mean ± standard deviation of fold-change for each group ( n = 6–10). Statistical significance defined as * p

    Journal: Stem Cell Research & Therapy

    Article Title: Responses to altered oxygen tension are distinct between human stem cells of high and low chondrogenic capacity

    doi: 10.1186/s13287-016-0419-8

    Figure Lengend Snippet: Gene expression analysis for fold-change of chondrogenic markers of the articular cartilage phenotype ( COL2A1 , ACAN ), the fibrocartilaginous phenotype ( COL1A1 ), and the hypertrophic phenotype ( COL10A1 , MMP13 ) demonstrates varied chondrogenic responses by high-GAG and low-GAG groups of each cell type, MSCs and ACPs, during pellet culture in physioxic relative to hyperoxic conditions. Data are mean ± standard deviation of fold-change for each group ( n = 6–10). Statistical significance defined as * p

    Article Snippet: Pellet cultures were formed by centrifuging 5 × 104 cells (MSCs) or 1 × 105 cells (ACPs) at 500 × g for 5 min in 240 μl of medium in Nunc polypropylene V-bottom 96-well plates (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Expressing, Standard Deviation

    Differential gene expression by Ad-MSC versus BM-MSC. The Partek Genomics Suite was used to import microarray data from the canine 1.0 ST Affymetrix chip. (A) A principle component analysis plot generated from these data depicts three biological replicates of each cell type, with red representing Ad-MSC, and blue representing BM-MSC. An ellipsoid was drawn around cell types using a sample standard deviation of 2. PC1 depicts a variation of 36.3%, PC2 a variation of 22.4%, and PC3 a variation of 5.6%. (B) Volcano plot of all differentially expressed genes in the analysis of Ad-MSC versus BM-MSC. x-Axis represents fold change, thick axis lines drawn at fold change −2 and 2. y-Axis represents P value, thick axis line marks 0.02. Color code represents column number assigned to each gene location on the Affymetrix chip. (C) section. Red represents a higher expression, and blue represents lower expression with scale bar for each category (Ad-MSC or BM-MSC) on the left side of plots.

    Journal: Stem Cells and Development

    Article Title: Mechanisms of Immune Suppression Utilized by Canine Adipose and Bone Marrow-Derived Mesenchymal Stem Cells

    doi: 10.1089/scd.2016.0207

    Figure Lengend Snippet: Differential gene expression by Ad-MSC versus BM-MSC. The Partek Genomics Suite was used to import microarray data from the canine 1.0 ST Affymetrix chip. (A) A principle component analysis plot generated from these data depicts three biological replicates of each cell type, with red representing Ad-MSC, and blue representing BM-MSC. An ellipsoid was drawn around cell types using a sample standard deviation of 2. PC1 depicts a variation of 36.3%, PC2 a variation of 22.4%, and PC3 a variation of 5.6%. (B) Volcano plot of all differentially expressed genes in the analysis of Ad-MSC versus BM-MSC. x-Axis represents fold change, thick axis lines drawn at fold change −2 and 2. y-Axis represents P value, thick axis line marks 0.02. Color code represents column number assigned to each gene location on the Affymetrix chip. (C) section. Red represents a higher expression, and blue represents lower expression with scale bar for each category (Ad-MSC or BM-MSC) on the left side of plots.

    Article Snippet: To compare the gene expression patterns of Ad-MSC and BM-MSC, microarray studies were done, using Affymetrix canine OST 1.0 chips. (The full array data have been deposited in the Gene Expression Omnibus database online.)

    Techniques: Expressing, Microarray, Chromatin Immunoprecipitation, Generated, Standard Deviation

    Sdf-1 impact on bone marrow mesenchymal stem cells. (A) Western blotting analysis of CXCR4, CD9, and tubulin in the whole population of bone marrow-derived mesenchymal stem cells (BM-MSCs) as well as of CXCR4 + and CXCR4 – BM-MSCs fractions fractions. (B) Quantitative RT-PCR analysis of CXCR4 and CD9 mRNA in BM-MSCs in control and Sdf-1-treated BM-MSCs. (C) Western blotting analysis of CD9 and tubulin in control and Sdf-1-treated (Sdf-1) BM-MSCs. (D) Migration of BM-MSCs in Sdf-1 gradient. The number of cells that migrated from the inserts was counted. (E) Percent of hybrid myotubes formed in co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs. (F) Co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs (green, localisation of β-galactosidase; blue, nuclei). Bar = 50 μm. CXCR, CXC chemokine receptor. * P

    Journal: Stem Cell Research & Therapy

    Article Title: Sdf-1 (CXCL12) induces CD9 expression in stem cells engaged in muscle regeneration

    doi: 10.1186/s13287-015-0041-1

    Figure Lengend Snippet: Sdf-1 impact on bone marrow mesenchymal stem cells. (A) Western blotting analysis of CXCR4, CD9, and tubulin in the whole population of bone marrow-derived mesenchymal stem cells (BM-MSCs) as well as of CXCR4 + and CXCR4 – BM-MSCs fractions fractions. (B) Quantitative RT-PCR analysis of CXCR4 and CD9 mRNA in BM-MSCs in control and Sdf-1-treated BM-MSCs. (C) Western blotting analysis of CD9 and tubulin in control and Sdf-1-treated (Sdf-1) BM-MSCs. (D) Migration of BM-MSCs in Sdf-1 gradient. The number of cells that migrated from the inserts was counted. (E) Percent of hybrid myotubes formed in co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs. (F) Co-culture of C2C12 myoblasts and control or Sdf-1 pretreated BM-MSCs (green, localisation of β-galactosidase; blue, nuclei). Bar = 50 μm. CXCR, CXC chemokine receptor. * P

    Article Snippet: Analyses of mRNA levels in BM-MSCs and ESCs included RNA isolation using the mirVana kit (Life Technologies) and then reverse transcription using Superscript (Life Technologies).

    Techniques: Western Blot, Derivative Assay, Quantitative RT-PCR, Migration, Co-Culture Assay

    rAMF increases the in vitro chemotaxis of MSCs towards HCC and their adhesion to endothelial cells. A) Pretreatment of BM-MSCs with 1 µg/ml rAMF (black bars) increases chemotaxis towards TCM derived from HuH7 or HC-PT-5 cells compared to untreated cells (white bars). B) Wound-healing assay of MSCs after pretreatment with rAMF or control (DMEM). Representative images were taken 24 hours after scratching. C) Adhesion to HMEC-1 endothelial cells was increased in BM-MSCs exposed to rAMF. D) Expression of AMF receptor (AMFR), GDP dissociation inhibitor 2 (GDI-1), caveolin-1 (CAV-1) and caveolin-2 (CAV-2) by qRT-PCR. *p

    Journal: PLoS ONE

    Article Title: Increased Migration of Human Mesenchymal Stromal Cells by Autocrine Motility Factor (AMF) Resulted in Enhanced Recruitment towards Hepatocellular Carcinoma

    doi: 10.1371/journal.pone.0095171

    Figure Lengend Snippet: rAMF increases the in vitro chemotaxis of MSCs towards HCC and their adhesion to endothelial cells. A) Pretreatment of BM-MSCs with 1 µg/ml rAMF (black bars) increases chemotaxis towards TCM derived from HuH7 or HC-PT-5 cells compared to untreated cells (white bars). B) Wound-healing assay of MSCs after pretreatment with rAMF or control (DMEM). Representative images were taken 24 hours after scratching. C) Adhesion to HMEC-1 endothelial cells was increased in BM-MSCs exposed to rAMF. D) Expression of AMF receptor (AMFR), GDP dissociation inhibitor 2 (GDI-1), caveolin-1 (CAV-1) and caveolin-2 (CAV-2) by qRT-PCR. *p

    Article Snippet: For in vivo migration studies, BM-MSCs or BM-MSCs pretreated with rAMF (5×105 ) were prelabeled with CMDiI for histological analysis and DiR (Molecular Probes, Invitrogen) for fluorescence imaging (FI).

    Techniques: In Vitro, Chemotaxis Assay, Derivative Assay, Wound Healing Assay, Expressing, Quantitative RT-PCR

    rAMF increases the in vivo migration and anchorage of MSCs to HCC tumors. BM-MSCs prestimulated with 1 µg/ml of rAMF were labeled with DiR and CMDiI cell trackers and IV injected in SC HuH7 tumor-bearing mice. After 3 days, tumors were removed and exposed to FI. A) Total FI was calculated by measuring the region of interest (ROI) for all the tissues isolated and the results were expressed as total radiant efficiency. ns, non significant. B) Representative tumor images of mice inoculated with rAMF-prestimulated BM-MSCs (MSC-rAMF) or unstimulated cells (MSCs). Images represent the average radiant efficiency. Region of interest (ROI) was calculated for the isolated tumor (C), liver (D), lung (E) and spleen (F) and the results were expressed as the average radiant efficiency. **p

    Journal: PLoS ONE

    Article Title: Increased Migration of Human Mesenchymal Stromal Cells by Autocrine Motility Factor (AMF) Resulted in Enhanced Recruitment towards Hepatocellular Carcinoma

    doi: 10.1371/journal.pone.0095171

    Figure Lengend Snippet: rAMF increases the in vivo migration and anchorage of MSCs to HCC tumors. BM-MSCs prestimulated with 1 µg/ml of rAMF were labeled with DiR and CMDiI cell trackers and IV injected in SC HuH7 tumor-bearing mice. After 3 days, tumors were removed and exposed to FI. A) Total FI was calculated by measuring the region of interest (ROI) for all the tissues isolated and the results were expressed as total radiant efficiency. ns, non significant. B) Representative tumor images of mice inoculated with rAMF-prestimulated BM-MSCs (MSC-rAMF) or unstimulated cells (MSCs). Images represent the average radiant efficiency. Region of interest (ROI) was calculated for the isolated tumor (C), liver (D), lung (E) and spleen (F) and the results were expressed as the average radiant efficiency. **p

    Article Snippet: For in vivo migration studies, BM-MSCs or BM-MSCs pretreated with rAMF (5×105 ) were prelabeled with CMDiI for histological analysis and DiR (Molecular Probes, Invitrogen) for fluorescence imaging (FI).

    Techniques: In Vivo, Migration, Labeling, Injection, Mouse Assay, Isolation

    AMF potently stimulates in vitro chemotaxis of MSCs from different sources. A) Detection of AMF (55 kDa) by western blot in CCM derived from HCC cells and TCM from ex vivo HCC SC tumors (upper panel). Colloidal Coomassie staining was performed as loading control (lower panel). MSC migration was analyzed with a Boyden chamber assay using rAMF as chemoattractant for BM-MSCs (B), HUCPVCs (C) or AT-MSCs (D). Results were expressed as percentage of control (DMEM) ±SEM. *p

    Journal: PLoS ONE

    Article Title: Increased Migration of Human Mesenchymal Stromal Cells by Autocrine Motility Factor (AMF) Resulted in Enhanced Recruitment towards Hepatocellular Carcinoma

    doi: 10.1371/journal.pone.0095171

    Figure Lengend Snippet: AMF potently stimulates in vitro chemotaxis of MSCs from different sources. A) Detection of AMF (55 kDa) by western blot in CCM derived from HCC cells and TCM from ex vivo HCC SC tumors (upper panel). Colloidal Coomassie staining was performed as loading control (lower panel). MSC migration was analyzed with a Boyden chamber assay using rAMF as chemoattractant for BM-MSCs (B), HUCPVCs (C) or AT-MSCs (D). Results were expressed as percentage of control (DMEM) ±SEM. *p

    Article Snippet: For in vivo migration studies, BM-MSCs or BM-MSCs pretreated with rAMF (5×105 ) were prelabeled with CMDiI for histological analysis and DiR (Molecular Probes, Invitrogen) for fluorescence imaging (FI).

    Techniques: In Vitro, Chemotaxis Assay, Western Blot, Derivative Assay, Ex Vivo, Staining, Migration, Boyden Chamber Assay

    MMP3 expression and MMP2 activity is induced in MSCs by rAMF. A) Analysis of MMP3 expression by qRT-PCR in BM-MSCs (black bars), HUCPVCs (white bars) or AT-MSCs (gray bars) stimulated with 1 µg/ml of rAMF. **p

    Journal: PLoS ONE

    Article Title: Increased Migration of Human Mesenchymal Stromal Cells by Autocrine Motility Factor (AMF) Resulted in Enhanced Recruitment towards Hepatocellular Carcinoma

    doi: 10.1371/journal.pone.0095171

    Figure Lengend Snippet: MMP3 expression and MMP2 activity is induced in MSCs by rAMF. A) Analysis of MMP3 expression by qRT-PCR in BM-MSCs (black bars), HUCPVCs (white bars) or AT-MSCs (gray bars) stimulated with 1 µg/ml of rAMF. **p

    Article Snippet: For in vivo migration studies, BM-MSCs or BM-MSCs pretreated with rAMF (5×105 ) were prelabeled with CMDiI for histological analysis and DiR (Molecular Probes, Invitrogen) for fluorescence imaging (FI).

    Techniques: Expressing, Activity Assay, Quantitative RT-PCR