primary human bone marrow derived mscs  (Lonza)


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    Name:
    hMSC Human Mesenchymal Stem Cells
    Description:
    Cryopreserved ampule of Human Mesenchymal Stem Cells hMSCs containing 750 000 cells
    Catalog Number:
    PT-2501
    Price:
    None
    Category:
    Primary and Stem Cells
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    Structured Review

    Lonza primary human bone marrow derived mscs
    Long-term monitoring of human <t>MSCs</t> in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 <t>hBM-MSC</t> or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment
    Cryopreserved ampule of Human Mesenchymal Stem Cells hMSCs containing 750 000 cells
    https://www.bioz.com/result/primary human bone marrow derived mscs/product/Lonza
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary human bone marrow derived mscs - by Bioz Stars, 2021-06
    97/100 stars

    Images

    1) Product Images from "Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration"

    Article Title: Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-1076-x

    Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment
    Figure Legend Snippet: Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment

    Techniques Used: Mouse Assay, Ex Vivo, Imaging

    2) Product Images from "Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration"

    Article Title: Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-1076-x

    Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment
    Figure Legend Snippet: Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment

    Techniques Used: Mouse Assay, Ex Vivo, Imaging

    Related Articles

    Cell Culture:

    Article Title: Human limbal niche cells are a powerful regenerative source for the prevention of limbal stem cell deficiency in a rabbit model
    Article Snippet: .. The second passage of bone marrow-derived MSC (BMMSC, PT-2501) was obtained from LONZA (Allendale, NJ) and cultured in parallel. .. Three dimensional (3D) Matrigel was prepared by adding 150 μl of 50% Matrigel (diluted in MESCM) per chamber of an 8-well chamber slide following incubation at 37 °C for 1 h. Cells were seeded on 3D Matrigel and cultured for 10 days in MESCM.

    Article Title: Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration
    Article Snippet: .. Cell preparation Mouse kidney-derived stem cells (mKSCs) [ ], the D1 mouse MSC (mMSC) line (D1 ORL UVA [D1](ATCC® CRL-12424™)), primary human umbilical cord-derived MSCs (hUC-MSCs; collected from consenting donors and produced identically to those already being used in clinical trials by NHS Blood and Transplant (NHSBT)), primary human bone marrow-derived MSCs (hBM-MSCs; Lonza PT-2501), human kidney cells (hKCs; the kidneys deemed unsuitable for transplantation via UK NHSBT [ ]) and RAW264.7 macrophages (European Collection of Authenticated Cell Cultures 91062702) were cultured at 37 °C under a humidified atmosphere with 5% CO2 (culture media are described in Additional file ). .. Primary human cells were used up to passage 8, whereas mouse lines were cultured up to passage 25.

    Article Title: The Effect of Antidepressants on Mesenchymal Stem Cell Differentiation
    Article Snippet: .. Human MSC (hMSC) were obtained commercially from Lonza (Basel, Switzerland), and cultured as previously described. .. [ , ] Briefly, cells were plated in six-well culture dishes containing MSC growth media (MSCGM; Lonza), and grown at 37℃ in a humidified atmosphere containing 5% CO2 with media changes every 3 to 4 days.

    Article Title: Therapeutic Use of Stem Cell Transplantation for Cell Replacement or Cytoprotective Effect of Microvesicle Released from Mesenchymal Stem Cell
    Article Snippet: In this respect, we try to examine the two possible mechanisms of pulmonary function restoration after human mesenchymal stem cell (hMSCs) transplantation by examining direct differentiation of transferred hMSCs into lung cell using human sequence specific RT-PCR and checking the effect of stem cell-derived microvesicles. .. Human mesenchymal stem cells and microvesicle preparation Bone marrow-derived human mesenchymal stem cells (hMSCs) were purchased from Lonza (Switzerland) and cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM) containing 20% fetal bovine serum (Gibco, Invitrogen, USA) at 37°C in a 5% CO2 atmosphere ( ). .. Microvesicles were isolated from conditioned media of hMSCs by using the ExoQuick (SBI Biosciences, USA).

    Produced:

    Article Title: Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration
    Article Snippet: .. Cell preparation Mouse kidney-derived stem cells (mKSCs) [ ], the D1 mouse MSC (mMSC) line (D1 ORL UVA [D1](ATCC® CRL-12424™)), primary human umbilical cord-derived MSCs (hUC-MSCs; collected from consenting donors and produced identically to those already being used in clinical trials by NHS Blood and Transplant (NHSBT)), primary human bone marrow-derived MSCs (hBM-MSCs; Lonza PT-2501), human kidney cells (hKCs; the kidneys deemed unsuitable for transplantation via UK NHSBT [ ]) and RAW264.7 macrophages (European Collection of Authenticated Cell Cultures 91062702) were cultured at 37 °C under a humidified atmosphere with 5% CO2 (culture media are described in Additional file ). .. Primary human cells were used up to passage 8, whereas mouse lines were cultured up to passage 25.

    Transplantation Assay:

    Article Title: Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration
    Article Snippet: .. Cell preparation Mouse kidney-derived stem cells (mKSCs) [ ], the D1 mouse MSC (mMSC) line (D1 ORL UVA [D1](ATCC® CRL-12424™)), primary human umbilical cord-derived MSCs (hUC-MSCs; collected from consenting donors and produced identically to those already being used in clinical trials by NHS Blood and Transplant (NHSBT)), primary human bone marrow-derived MSCs (hBM-MSCs; Lonza PT-2501), human kidney cells (hKCs; the kidneys deemed unsuitable for transplantation via UK NHSBT [ ]) and RAW264.7 macrophages (European Collection of Authenticated Cell Cultures 91062702) were cultured at 37 °C under a humidified atmosphere with 5% CO2 (culture media are described in Additional file ). .. Primary human cells were used up to passage 8, whereas mouse lines were cultured up to passage 25.

    other:

    Article Title: Enhanced MSC Chondrogenesis Following Delivery of TGF-?3 from Alginate Microspheres within Hyaluronic Acid Hydrogels In Vitro and In Vivo
    Article Snippet: To evaluate the efficacy of released TGF-β3 in promoting chondrogenesis, human MSCs in HA hydrogels were assessed through day 28.

    Derivative Assay:

    Article Title: Targeting Cellular Metabolism in Acute Myeloid Leukemia and the Role of Patient Heterogeneity
    Article Snippet: .. Cocultures of Human Mesenchymal Stem Cells (MSCs) and AML CellsHuman MSCs (Lonza, Cambrex BioScience, Walkersville, MD, USA) derived from a healthy donor (MSC24539) were expanded in complete MSC growth medium (MSCGM™; Lonza) with 10% heat-inactivated FBS and 4 mM L-glutamine. ..

    Immunofluorescence:

    Article Title: A Contact-Based Method for Differentiation of Human Mesenchymal Stem Cells into an Endothelial Cell-Phenotype
    Article Snippet: Phase-contrast imaging was performed again after 72 h and at the end of 7 days of culture (37 °C, 5% CO2 and 95% RH). .. To detect differentiation of human MSC into an EC-type, immunofluorescence detection for CD31 (or PECAM-1), an integral membrane glycoprotein that is expressed at high levels on endothelial cells, was performed [ ]. .. For the immunostaining, a primary anti-CD31 monoclonal antibody and a secondary fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG (Abcam, Cambridge, UK) was used in immunofluorescence detection of the CD31 marker expression in human MSC cultured atop various substrates (including Plate-1, Plate-2, and controls).

    Modification:

    Article Title: Therapeutic Use of Stem Cell Transplantation for Cell Replacement or Cytoprotective Effect of Microvesicle Released from Mesenchymal Stem Cell
    Article Snippet: In this respect, we try to examine the two possible mechanisms of pulmonary function restoration after human mesenchymal stem cell (hMSCs) transplantation by examining direct differentiation of transferred hMSCs into lung cell using human sequence specific RT-PCR and checking the effect of stem cell-derived microvesicles. .. Human mesenchymal stem cells and microvesicle preparation Bone marrow-derived human mesenchymal stem cells (hMSCs) were purchased from Lonza (Switzerland) and cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM) containing 20% fetal bovine serum (Gibco, Invitrogen, USA) at 37°C in a 5% CO2 atmosphere ( ). .. Microvesicles were isolated from conditioned media of hMSCs by using the ExoQuick (SBI Biosciences, USA).

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  • 98
    Lonza bone marrow derived msc
    Expression of CD73, CD90, CD105 in LNC and <t>BMMSC.</t> P4 LNC and P4 BMMSC cultured on 2D Matrigel in MESCM were subjected to fluorescence-activated cell sorting (FACS) analysis of <t>MSC</t> markers ( A – F , n = 3).
    Bone Marrow Derived Msc, supplied by Lonza, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bone marrow derived msc/product/Lonza
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bone marrow derived msc - by Bioz Stars, 2021-06
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    96
    Lonza primary cells hbm mscs
    Effects of PZR on <t>hBM</t> <t>MSCs</t> in regulating adhesion to and spreading and migration on ECM molecules. ( A ) BCECF-labeled hBM MSC adhesion to BSA, FN, VN, COL-I, COL-IV, and LN after 30 min incubation. ( B ) hBM MSC spreading on FN, VN, COL-I, COL-IV, LN, or BSA for 60 min, then phase contrast images taken and percentages of spreading cells calculated. Cells adhering or spreading were calculated as the mean percentage + S.E.M. of input cells. ( Ci ) hBM MSC migrating on FN, VN, COL-I, COL-IV, LN, or BSA for 20 h. Left: Cell migration on BSA was normalized to 100% and percentage increase in migration on ECM substrates compared to this normalized value. ( Cii ) Representative phase contrast images of hBM MSC migration at 0 (left panel) and 20 h (right panel) after initiating the migration assay. Black dashed lines mark the migratory area. t = hours after initiation of migration. ( D – F ) Respective adhesion, spreading, and migration assays were repeated after hBM MSCs were treated with PZR2-, PZR3-, PZR4 -, and PZR1- or control siRNAs (see Section 2.6 ). In ( F ), hBM MSCs treated with PZR and/or PZRb siRNAs were compared to control siRNA transfected cells, which were normalized to 100%. Representative phase contrast images of hBM MSC spreading (Inset E ) and migration at 0 (left panel) and 20 h (right panel) after initiating the migration assay (Inset F ) following knockdown with control, PZR2, and PZR3 siRNAs. Black dashed lines mark the migratory area. t = hours after initiation of migration. ( G – I ) Respective adhesion, spreading, and migration assays were repeated after hBM MSCs were treated with NEDD9.1 - or NEDD9.2 - or control siRNAs (see Supplementary Materials and Methods ). In ( G ), hBM MSCs treated with NEDD9.1 - or NEDD9.2 -siRNAs were compared to control siRNA transfected cells, which were normalized to 100%. Sham transfected cells lacking specific siRNAs were also tested. Values are means ± S.E.M. ( n = 3 independent experiments using three different batches of hBM MSCs).
    Primary Cells Hbm Mscs, supplied by Lonza, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary cells hbm mscs/product/Lonza
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    94
    Lonza human primary bone marrow mesenchymal stem cells
    Derivation of iMSCs and the comparison of molecular signatures. ( A ) The experimental scheme for efficient derivation of iMSCs from dermal fibroblasts with 6 chemicals with or without LIF, TGF-β, and bFGF. Expanded iMSCs with clonal expansion ability (clonogenicity) can be further differentiated into different lineages (multipotency) or treat sepsis-induced acute lung injury in the mouse model. ( B ) The representative flow cytometry analysis of <t>human</t> dermal fibroblasts CRL2097, iMSCs, and BMMSCs with MSC functional markers SSEA-4 and PODXL. SSEA-4 and PODXL were abundantly expressed in iMSCs and BMMSCs, but not in fibroblasts. ( C ) Clonogenicity of fibroblasts, iMSCs, and BMMSCs. By a 96-well colony formation assay, quantitation of CFU-F in fibroblasts (1 per 112 <t>cells),</t> iMSCs (1 per 7 cells), and BMMSCs (1 per 22 cells) was obtained by the negative linear relationship between the number of seeded cells and the percentage of wells with no colonies. ( D ) Flow cytometry analysis of OCT4 protein expression in human fibroblasts, iMSCs, BMMSCs, and human embryonic <t>stem</t> cells (hESCs). The pluripotent marker OCT4 was at a basal level in fibroblasts (CRL2097), slightly up-regulated in iMSCs and BMMSCs, and highly expressed in hESCs. ( E ) The principal component analysis of the expression of stemness genes in three different dermal fibroblasts (CRL2097, DF440547, and DF443480), iMSCs induced from the three different fibroblasts, and two independent sources of BMMSCs (BMMSC_1: <t>primary</t> human <t>bone</t> <t>marrow</t> MSCs used throughout this study; BMMSC_2: publicly available gene expression data for human BMMSCs with accession number GSM1533333). Expression probes were functionally annotated with Gene Ontology (GO) and selected by text-mining for the term “stem cell maintenance” in R (418 probes). Principal component 1 accounts for 40%, principal component 2 accounts for 19%, and principal component 3 accounts for 14% of the variation in the dataset. The clustering of iMSCs derived from three different fibroblast sources between two different BMMSCs suggests the robust efficacy of the cocktail.
    Human Primary Bone Marrow Mesenchymal Stem Cells, supplied by Lonza, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    Image Search Results


    Expression of CD73, CD90, CD105 in LNC and BMMSC. P4 LNC and P4 BMMSC cultured on 2D Matrigel in MESCM were subjected to fluorescence-activated cell sorting (FACS) analysis of MSC markers ( A – F , n = 3).

    Journal: Scientific Reports

    Article Title: Human limbal niche cells are a powerful regenerative source for the prevention of limbal stem cell deficiency in a rabbit model

    doi: 10.1038/s41598-018-24862-6

    Figure Lengend Snippet: Expression of CD73, CD90, CD105 in LNC and BMMSC. P4 LNC and P4 BMMSC cultured on 2D Matrigel in MESCM were subjected to fluorescence-activated cell sorting (FACS) analysis of MSC markers ( A – F , n = 3).

    Article Snippet: The second passage of bone marrow-derived MSC (BMMSC, PT-2501) was obtained from LONZA (Allendale, NJ) and cultured in parallel.

    Techniques: Expressing, Cell Culture, Fluorescence, FACS

    LNC express more ESC, MSC and NC markers. P4 LNC and P4 BMMSC cultured on 2D Matrigel in MESCM were subjected to qRT-PCR for transcription expression of ESC markers ( A ), MSC and neural crest markers ( B , n = 3, *p

    Journal: Scientific Reports

    Article Title: Human limbal niche cells are a powerful regenerative source for the prevention of limbal stem cell deficiency in a rabbit model

    doi: 10.1038/s41598-018-24862-6

    Figure Lengend Snippet: LNC express more ESC, MSC and NC markers. P4 LNC and P4 BMMSC cultured on 2D Matrigel in MESCM were subjected to qRT-PCR for transcription expression of ESC markers ( A ), MSC and neural crest markers ( B , n = 3, *p

    Article Snippet: The second passage of bone marrow-derived MSC (BMMSC, PT-2501) was obtained from LONZA (Allendale, NJ) and cultured in parallel.

    Techniques: Cell Culture, Quantitative RT-PCR, Expressing

    Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment

    Journal: Stem Cell Research & Therapy

    Article Title: Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration

    doi: 10.1186/s13287-018-1076-x

    Figure Lengend Snippet: Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment

    Article Snippet: Cell preparation Mouse kidney-derived stem cells (mKSCs) [ ], the D1 mouse MSC (mMSC) line (D1 ORL UVA [D1](ATCC® CRL-12424™)), primary human umbilical cord-derived MSCs (hUC-MSCs; collected from consenting donors and produced identically to those already being used in clinical trials by NHS Blood and Transplant (NHSBT)), primary human bone marrow-derived MSCs (hBM-MSCs; Lonza PT-2501), human kidney cells (hKCs; the kidneys deemed unsuitable for transplantation via UK NHSBT [ ]) and RAW264.7 macrophages (European Collection of Authenticated Cell Cultures 91062702) were cultured at 37 °C under a humidified atmosphere with 5% CO2 (culture media are described in Additional file ).

    Techniques: Mouse Assay, Ex Vivo, Imaging

    Effects of PZR on hBM MSCs in regulating adhesion to and spreading and migration on ECM molecules. ( A ) BCECF-labeled hBM MSC adhesion to BSA, FN, VN, COL-I, COL-IV, and LN after 30 min incubation. ( B ) hBM MSC spreading on FN, VN, COL-I, COL-IV, LN, or BSA for 60 min, then phase contrast images taken and percentages of spreading cells calculated. Cells adhering or spreading were calculated as the mean percentage + S.E.M. of input cells. ( Ci ) hBM MSC migrating on FN, VN, COL-I, COL-IV, LN, or BSA for 20 h. Left: Cell migration on BSA was normalized to 100% and percentage increase in migration on ECM substrates compared to this normalized value. ( Cii ) Representative phase contrast images of hBM MSC migration at 0 (left panel) and 20 h (right panel) after initiating the migration assay. Black dashed lines mark the migratory area. t = hours after initiation of migration. ( D – F ) Respective adhesion, spreading, and migration assays were repeated after hBM MSCs were treated with PZR2-, PZR3-, PZR4 -, and PZR1- or control siRNAs (see Section 2.6 ). In ( F ), hBM MSCs treated with PZR and/or PZRb siRNAs were compared to control siRNA transfected cells, which were normalized to 100%. Representative phase contrast images of hBM MSC spreading (Inset E ) and migration at 0 (left panel) and 20 h (right panel) after initiating the migration assay (Inset F ) following knockdown with control, PZR2, and PZR3 siRNAs. Black dashed lines mark the migratory area. t = hours after initiation of migration. ( G – I ) Respective adhesion, spreading, and migration assays were repeated after hBM MSCs were treated with NEDD9.1 - or NEDD9.2 - or control siRNAs (see Supplementary Materials and Methods ). In ( G ), hBM MSCs treated with NEDD9.1 - or NEDD9.2 -siRNAs were compared to control siRNA transfected cells, which were normalized to 100%. Sham transfected cells lacking specific siRNAs were also tested. Values are means ± S.E.M. ( n = 3 independent experiments using three different batches of hBM MSCs).

    Journal: Cells

    Article Title: P0-Related Protein Accelerates Human Mesenchymal Stromal Cell Migration by Modulating VLA-5 Interactions with Fibronectin

    doi: 10.3390/cells9051100

    Figure Lengend Snippet: Effects of PZR on hBM MSCs in regulating adhesion to and spreading and migration on ECM molecules. ( A ) BCECF-labeled hBM MSC adhesion to BSA, FN, VN, COL-I, COL-IV, and LN after 30 min incubation. ( B ) hBM MSC spreading on FN, VN, COL-I, COL-IV, LN, or BSA for 60 min, then phase contrast images taken and percentages of spreading cells calculated. Cells adhering or spreading were calculated as the mean percentage + S.E.M. of input cells. ( Ci ) hBM MSC migrating on FN, VN, COL-I, COL-IV, LN, or BSA for 20 h. Left: Cell migration on BSA was normalized to 100% and percentage increase in migration on ECM substrates compared to this normalized value. ( Cii ) Representative phase contrast images of hBM MSC migration at 0 (left panel) and 20 h (right panel) after initiating the migration assay. Black dashed lines mark the migratory area. t = hours after initiation of migration. ( D – F ) Respective adhesion, spreading, and migration assays were repeated after hBM MSCs were treated with PZR2-, PZR3-, PZR4 -, and PZR1- or control siRNAs (see Section 2.6 ). In ( F ), hBM MSCs treated with PZR and/or PZRb siRNAs were compared to control siRNA transfected cells, which were normalized to 100%. Representative phase contrast images of hBM MSC spreading (Inset E ) and migration at 0 (left panel) and 20 h (right panel) after initiating the migration assay (Inset F ) following knockdown with control, PZR2, and PZR3 siRNAs. Black dashed lines mark the migratory area. t = hours after initiation of migration. ( G – I ) Respective adhesion, spreading, and migration assays were repeated after hBM MSCs were treated with NEDD9.1 - or NEDD9.2 - or control siRNAs (see Supplementary Materials and Methods ). In ( G ), hBM MSCs treated with NEDD9.1 - or NEDD9.2 -siRNAs were compared to control siRNA transfected cells, which were normalized to 100%. Sham transfected cells lacking specific siRNAs were also tested. Values are means ± S.E.M. ( n = 3 independent experiments using three different batches of hBM MSCs).

    Article Snippet: Primary Cells hBM MSCs were purchased from Lonza Biologics, Slough, England at passage 2 and maintained in culture in mesenchymal stem cell growth medium (MSCGM, Lonza Biologics) supplemented with 10% FCS (Gibco-BRL, Thermo Fisher Scientific, Milton Keynes, England).

    Techniques: Migration, Labeling, Incubation, Transfection

    Differential human P 0 -related protein (PZR) and PZRb expression on hBM MSCs. ( A ) Q-RT-PCR of PZR and PZRb using 3 different hBM MSC batches (mean ± S.E.M.) and human PZR and PZRb primers and probes described in Section 2.5 . ( B ) Q-RT-PCR of PZR , PZRb , and the positive control VEGF transcripts in two independent batches of hBM MSC in normoxia (20% O 2 ) with or without 150 μM CoCl 2 or hypoxia (1.5% O 2 ) for 4, 16, and 24 h. Values are means ± S.E.M. of triplicate assays. ( C ) Flow cytometric histograms of human PZR and PZRb protein expression after mIgG1 (panels a , c , and e ) or WM78 (panels b , d , and f ) staining of MEF cells either untransfected ( a , b ) or expressing hPZR ( c , d ) or hPZRb ( e , f ) plus Alexa-488 goat anti-mouse IgG1 antibody. P2 is the gate set against the isotype control. The median fluorescence intensity (MFI) of cells in the positive gate is shown on each histogram. Values above are means ± S.E.M. for triplicate assays. ( D ) Representative FACS histogram of hBM MSCs staining with WM78 compared to the mIgG1 negative control stained as above. MFIs are means ± S.E.M. of three independent experiments using three different batches of hBM MSCs. ( E ) Immunoprecipitation of human PZR isoforms using WM78. Left. Biotinylated MEF cells expressing human PZRb (lanes A and B ) and human PZR (lanes C and D ) were immunoprecipitated with the mIgG1 (lanes A and D ) or WM78 (lanes B and C ) and then Western blotted using streptavidin-HRP. Right. Immunoprecipitation of human PZR isoforms from surface biotinylated hBM MSCs (lane A ) with WM78 or mIgG1 prior to electrophoresis and Western blotting using HRP-streptavidin.

    Journal: Cells

    Article Title: P0-Related Protein Accelerates Human Mesenchymal Stromal Cell Migration by Modulating VLA-5 Interactions with Fibronectin

    doi: 10.3390/cells9051100

    Figure Lengend Snippet: Differential human P 0 -related protein (PZR) and PZRb expression on hBM MSCs. ( A ) Q-RT-PCR of PZR and PZRb using 3 different hBM MSC batches (mean ± S.E.M.) and human PZR and PZRb primers and probes described in Section 2.5 . ( B ) Q-RT-PCR of PZR , PZRb , and the positive control VEGF transcripts in two independent batches of hBM MSC in normoxia (20% O 2 ) with or without 150 μM CoCl 2 or hypoxia (1.5% O 2 ) for 4, 16, and 24 h. Values are means ± S.E.M. of triplicate assays. ( C ) Flow cytometric histograms of human PZR and PZRb protein expression after mIgG1 (panels a , c , and e ) or WM78 (panels b , d , and f ) staining of MEF cells either untransfected ( a , b ) or expressing hPZR ( c , d ) or hPZRb ( e , f ) plus Alexa-488 goat anti-mouse IgG1 antibody. P2 is the gate set against the isotype control. The median fluorescence intensity (MFI) of cells in the positive gate is shown on each histogram. Values above are means ± S.E.M. for triplicate assays. ( D ) Representative FACS histogram of hBM MSCs staining with WM78 compared to the mIgG1 negative control stained as above. MFIs are means ± S.E.M. of three independent experiments using three different batches of hBM MSCs. ( E ) Immunoprecipitation of human PZR isoforms using WM78. Left. Biotinylated MEF cells expressing human PZRb (lanes A and B ) and human PZR (lanes C and D ) were immunoprecipitated with the mIgG1 (lanes A and D ) or WM78 (lanes B and C ) and then Western blotted using streptavidin-HRP. Right. Immunoprecipitation of human PZR isoforms from surface biotinylated hBM MSCs (lane A ) with WM78 or mIgG1 prior to electrophoresis and Western blotting using HRP-streptavidin.

    Article Snippet: Primary Cells hBM MSCs were purchased from Lonza Biologics, Slough, England at passage 2 and maintained in culture in mesenchymal stem cell growth medium (MSCGM, Lonza Biologics) supplemented with 10% FCS (Gibco-BRL, Thermo Fisher Scientific, Milton Keynes, England).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Staining, Fluorescence, FACS, Negative Control, Immunoprecipitation, Western Blot, Electrophoresis

    Co-localization of PZR and integrins on migrating NIH3T3 transfectants and hBM MSCs. ( Ai ) Representative FACS histograms of hBM MSCs stained for CD29 (β1 integrin), CD49e (α5 integrin), CD49d (α4 integrin), CD51 (vitronectin receptor), or the relevant isotype control (mIgG1) followed by Alexa-488 goat anti-mIgG1. MFI ± S.E.M. shown above histograms ( n = 3 independent experiments). Black histograms: integrin staining; white histograms: mIgG1 negative control. ( Aii ) hBM MSCs were untreated or incubated with blocking antibodies against CD29, CD49e, CD49d, CD51, and CD51/61 or the corresponding isotype controls before being allowed to adhere to fibronectin (FN) or the negative control, BSA. Values are means ± S.E.M. for three independent experiments performed in triplicate. ( B ) Migration assay using NIH3T3-hPZR ( a ) with one area analyzed by confocal microscopy circled in red. ( b,c ) NIH3T3-PZR transfectants from the 6 h migratory interface double stained with WM78 (PZR; green stain) and biotin-CD29 (red stain) or ( d ) biotin-CD49e (red stain) plus appropriate secondary fluorescent reagents. hBM MSCs from the 6 h migratory interface double stained with WM78 and Alexa488 goat anti mIgG1 (PZR; green stain), then blocked with mIgG1 and stained with biotinylated ( e ) CD29 (red stain) or ( f ) CD49e (red stain) with streptavidin-conjugated Alexa 546. There is a co-association of PZR with α5 (CD49e) or β1 (CD29) at the leading edge of the migrating cell. C ( a ) Quantitation of PZR co-localizing with CD29 and CD49e or CD49d and CD51 in 6 h migrating hBM MSCs. Values are means ± S.E.M. for three independent experiments performed in triplicate. Inset shows co-immuno-precipitation and Western blotting with respective the anti-PZR WM78 mAb and biotin conjugated anti-human CD49e or CD29, and using mIgG1 as the negative control for the immunoprecipitation (i.p.). Lane A: hBM MSC cell lysate, lane B: hBM MSC i.p with anti-PZR, lane C: hBM MSC i.p. with negative control. C ( b ) Confocal microscopy of PZR co-localizing with CD29 and CD49e and to a much lesser extent with CD49d and CD51 in NIH3T3-PZR after 6 h of migration. Inset shows co-immunoprecipitation and Western blotting of PZR (WM78 mAb) with biotin anti-mouse CD49e or CD29. mIgG1 was used as negative i.p. control. Lane A: NIH3T3-PZR cell lysate, lane B: NIH3T3-PZR i.p with WM78, lane C: NIH3T3-PZRb i.p. with WM78, lane D: NIH3T3 i.p. with WM78 and lane E: NIH3T3-hPZR i.p. with mIgG1.

    Journal: Cells

    Article Title: P0-Related Protein Accelerates Human Mesenchymal Stromal Cell Migration by Modulating VLA-5 Interactions with Fibronectin

    doi: 10.3390/cells9051100

    Figure Lengend Snippet: Co-localization of PZR and integrins on migrating NIH3T3 transfectants and hBM MSCs. ( Ai ) Representative FACS histograms of hBM MSCs stained for CD29 (β1 integrin), CD49e (α5 integrin), CD49d (α4 integrin), CD51 (vitronectin receptor), or the relevant isotype control (mIgG1) followed by Alexa-488 goat anti-mIgG1. MFI ± S.E.M. shown above histograms ( n = 3 independent experiments). Black histograms: integrin staining; white histograms: mIgG1 negative control. ( Aii ) hBM MSCs were untreated or incubated with blocking antibodies against CD29, CD49e, CD49d, CD51, and CD51/61 or the corresponding isotype controls before being allowed to adhere to fibronectin (FN) or the negative control, BSA. Values are means ± S.E.M. for three independent experiments performed in triplicate. ( B ) Migration assay using NIH3T3-hPZR ( a ) with one area analyzed by confocal microscopy circled in red. ( b,c ) NIH3T3-PZR transfectants from the 6 h migratory interface double stained with WM78 (PZR; green stain) and biotin-CD29 (red stain) or ( d ) biotin-CD49e (red stain) plus appropriate secondary fluorescent reagents. hBM MSCs from the 6 h migratory interface double stained with WM78 and Alexa488 goat anti mIgG1 (PZR; green stain), then blocked with mIgG1 and stained with biotinylated ( e ) CD29 (red stain) or ( f ) CD49e (red stain) with streptavidin-conjugated Alexa 546. There is a co-association of PZR with α5 (CD49e) or β1 (CD29) at the leading edge of the migrating cell. C ( a ) Quantitation of PZR co-localizing with CD29 and CD49e or CD49d and CD51 in 6 h migrating hBM MSCs. Values are means ± S.E.M. for three independent experiments performed in triplicate. Inset shows co-immuno-precipitation and Western blotting with respective the anti-PZR WM78 mAb and biotin conjugated anti-human CD49e or CD29, and using mIgG1 as the negative control for the immunoprecipitation (i.p.). Lane A: hBM MSC cell lysate, lane B: hBM MSC i.p with anti-PZR, lane C: hBM MSC i.p. with negative control. C ( b ) Confocal microscopy of PZR co-localizing with CD29 and CD49e and to a much lesser extent with CD49d and CD51 in NIH3T3-PZR after 6 h of migration. Inset shows co-immunoprecipitation and Western blotting of PZR (WM78 mAb) with biotin anti-mouse CD49e or CD29. mIgG1 was used as negative i.p. control. Lane A: NIH3T3-PZR cell lysate, lane B: NIH3T3-PZR i.p with WM78, lane C: NIH3T3-PZRb i.p. with WM78, lane D: NIH3T3 i.p. with WM78 and lane E: NIH3T3-hPZR i.p. with mIgG1.

    Article Snippet: Primary Cells hBM MSCs were purchased from Lonza Biologics, Slough, England at passage 2 and maintained in culture in mesenchymal stem cell growth medium (MSCGM, Lonza Biologics) supplemented with 10% FCS (Gibco-BRL, Thermo Fisher Scientific, Milton Keynes, England).

    Techniques: FACS, Staining, Negative Control, Incubation, Blocking Assay, Migration, Confocal Microscopy, Quantitation Assay, Immunoprecipitation, Western Blot

    Knockdown of hPZR shows reduced phosphorylation of FAK and reduced vinculin in migrating hBM MSCs. ( A ) Representative confocal microscopy images of PFAK (red) and DAPI (blue) staining in migrating hBM MSCs ( A ) without or ( B ) with fibronectin and treated with siRNAs control, PZR2 , PZR3 , and PZR4 . Original magnifications, 63×. ( C ) Quantification of PFAK expression in siRNA control treated hBM MSCs migrating without or fibronectin. The average integrated density of PFAK/the number of cells is shown as means ± S.E.M. per 100 cells counted (*** p

    Journal: Cells

    Article Title: P0-Related Protein Accelerates Human Mesenchymal Stromal Cell Migration by Modulating VLA-5 Interactions with Fibronectin

    doi: 10.3390/cells9051100

    Figure Lengend Snippet: Knockdown of hPZR shows reduced phosphorylation of FAK and reduced vinculin in migrating hBM MSCs. ( A ) Representative confocal microscopy images of PFAK (red) and DAPI (blue) staining in migrating hBM MSCs ( A ) without or ( B ) with fibronectin and treated with siRNAs control, PZR2 , PZR3 , and PZR4 . Original magnifications, 63×. ( C ) Quantification of PFAK expression in siRNA control treated hBM MSCs migrating without or fibronectin. The average integrated density of PFAK/the number of cells is shown as means ± S.E.M. per 100 cells counted (*** p

    Article Snippet: Primary Cells hBM MSCs were purchased from Lonza Biologics, Slough, England at passage 2 and maintained in culture in mesenchymal stem cell growth medium (MSCGM, Lonza Biologics) supplemented with 10% FCS (Gibco-BRL, Thermo Fisher Scientific, Milton Keynes, England).

    Techniques: Confocal Microscopy, Staining, Expressing

    Derivation of iMSCs and the comparison of molecular signatures. ( A ) The experimental scheme for efficient derivation of iMSCs from dermal fibroblasts with 6 chemicals with or without LIF, TGF-β, and bFGF. Expanded iMSCs with clonal expansion ability (clonogenicity) can be further differentiated into different lineages (multipotency) or treat sepsis-induced acute lung injury in the mouse model. ( B ) The representative flow cytometry analysis of human dermal fibroblasts CRL2097, iMSCs, and BMMSCs with MSC functional markers SSEA-4 and PODXL. SSEA-4 and PODXL were abundantly expressed in iMSCs and BMMSCs, but not in fibroblasts. ( C ) Clonogenicity of fibroblasts, iMSCs, and BMMSCs. By a 96-well colony formation assay, quantitation of CFU-F in fibroblasts (1 per 112 cells), iMSCs (1 per 7 cells), and BMMSCs (1 per 22 cells) was obtained by the negative linear relationship between the number of seeded cells and the percentage of wells with no colonies. ( D ) Flow cytometry analysis of OCT4 protein expression in human fibroblasts, iMSCs, BMMSCs, and human embryonic stem cells (hESCs). The pluripotent marker OCT4 was at a basal level in fibroblasts (CRL2097), slightly up-regulated in iMSCs and BMMSCs, and highly expressed in hESCs. ( E ) The principal component analysis of the expression of stemness genes in three different dermal fibroblasts (CRL2097, DF440547, and DF443480), iMSCs induced from the three different fibroblasts, and two independent sources of BMMSCs (BMMSC_1: primary human bone marrow MSCs used throughout this study; BMMSC_2: publicly available gene expression data for human BMMSCs with accession number GSM1533333). Expression probes were functionally annotated with Gene Ontology (GO) and selected by text-mining for the term “stem cell maintenance” in R (418 probes). Principal component 1 accounts for 40%, principal component 2 accounts for 19%, and principal component 3 accounts for 14% of the variation in the dataset. The clustering of iMSCs derived from three different fibroblast sources between two different BMMSCs suggests the robust efficacy of the cocktail.

    Journal: Scientific Reports

    Article Title: Efficient Generation of Chemically Induced Mesenchymal Stem Cells from Human Dermal Fibroblasts

    doi: 10.1038/srep44534

    Figure Lengend Snippet: Derivation of iMSCs and the comparison of molecular signatures. ( A ) The experimental scheme for efficient derivation of iMSCs from dermal fibroblasts with 6 chemicals with or without LIF, TGF-β, and bFGF. Expanded iMSCs with clonal expansion ability (clonogenicity) can be further differentiated into different lineages (multipotency) or treat sepsis-induced acute lung injury in the mouse model. ( B ) The representative flow cytometry analysis of human dermal fibroblasts CRL2097, iMSCs, and BMMSCs with MSC functional markers SSEA-4 and PODXL. SSEA-4 and PODXL were abundantly expressed in iMSCs and BMMSCs, but not in fibroblasts. ( C ) Clonogenicity of fibroblasts, iMSCs, and BMMSCs. By a 96-well colony formation assay, quantitation of CFU-F in fibroblasts (1 per 112 cells), iMSCs (1 per 7 cells), and BMMSCs (1 per 22 cells) was obtained by the negative linear relationship between the number of seeded cells and the percentage of wells with no colonies. ( D ) Flow cytometry analysis of OCT4 protein expression in human fibroblasts, iMSCs, BMMSCs, and human embryonic stem cells (hESCs). The pluripotent marker OCT4 was at a basal level in fibroblasts (CRL2097), slightly up-regulated in iMSCs and BMMSCs, and highly expressed in hESCs. ( E ) The principal component analysis of the expression of stemness genes in three different dermal fibroblasts (CRL2097, DF440547, and DF443480), iMSCs induced from the three different fibroblasts, and two independent sources of BMMSCs (BMMSC_1: primary human bone marrow MSCs used throughout this study; BMMSC_2: publicly available gene expression data for human BMMSCs with accession number GSM1533333). Expression probes were functionally annotated with Gene Ontology (GO) and selected by text-mining for the term “stem cell maintenance” in R (418 probes). Principal component 1 accounts for 40%, principal component 2 accounts for 19%, and principal component 3 accounts for 14% of the variation in the dataset. The clustering of iMSCs derived from three different fibroblast sources between two different BMMSCs suggests the robust efficacy of the cocktail.

    Article Snippet: Human primary bone marrow mesenchymal stem cells (BMMSCs) were purchased from LONZA and were cultured with Dulbecco’s Modified Eagle Media-low glucose (DMEM-LG) medium containing 10% FBS.

    Techniques: Flow Cytometry, Cytometry, Functional Assay, Colony Assay, Quantitation Assay, Expressing, Marker, Derivative Assay