mesenchymal stem cell basal medium  (ATCC)


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    Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs
    Description:
    Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs is a sterile phenol red free liquid tissue culture medium intended for use as one component in a complete ATCC Primary Cell Solutions system This low serum 2 FBS system is designed to support mesenchymal stem cells derived from various normal human tissues including lipoaspirates and umbilical cord Mesenchymal Stem Cell Basal Medium contains essential and non essential amino acids vitamins other organic compounds trace minerals and inorganic salts To support the proliferation and plating efficiency of various types of adult stem cells Mesenchymal Stem Cell Basal Medium must be supplemented with the appropriate cell specific growth kit When using this complete media system the growth of undifferentiated mesenchymal stem cells is supported without the use of feeder layers extracellular matrix proteins or other substrates For mesenchymal stem cells derived from human lipoaspirates e g Adipose Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 011 or umbilical cord Umbilical Cord Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 010 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Adipose and Umbilical derived MSCs Low serum ATCC PCS 500 040 For mesenchymal stem cells derived from bone marrow Bone Marrow derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 012 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Bone Marrow derived MSCs ATCC PCS 500 041 Optional media supplements Gentamicin Amphotericin B Solution ATCC PCS 999 025 Penicillin Streptomycin Amphotericin B Solution ATCC PCS 999 002 Phenol Red ATCC PCS 999 001
    Catalog Number:
    PCS-500-030
    Price:
    None
    Applications:
    Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs is a sterile, phenol red-free, liquid tissue culture medium intended for use as one component in a complete ATCC® Primary Cell Solutions™ system. This low serum (2% FBS) system is designed to support mesenchymal stem cells derived from various normal human tissues including lipoaspirates and umbilical cord. Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs contains essential and non-essential amino acids, vitamins, other organic compounds, trace minerals and inorganic salts. To support the proliferation and plating efficiency of various types of adult stem cells, Mesenchymal Stem Cell Basal Medium must be supplemented with the appropriate cell-specific growth kit. When using this complete media system, the growth of undifferentiated mesenchymal stem cells is supported without the use of feeder layers, extracellular matrix proteins or other substrates.  For mesenchymal stem cells derived from human lipoaspirates (e.g., Adipose-Derived Mesenchymal Stem Cells; Normal, Human, ATCC PCS-500-011) or umbilical cord (Umbilical Cord-Derived Mesenchymal Stem Cells; Normal, Human, ATCC PCS-500-010), supplement Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs with the Mesenchymal Stem Cell Growth Kit for Adipose and Umbilical-derived MSCs –Low serum (ATCC PCS-500-040). For mesenchymal stem cells derived from bone marrow (Bone Marrow-derived Mesenchymal Stem Cells; Normal, Human, ATCC PCS-500-012) supplement Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs with the Mesenchymal  Stem Cell Growth Kit for Bone Marrow-derived MSCs (ATCC PCS-500-041) Optional media supplements: Gentamicin-Amphotericin B Solution (ATCC PCS-999-025) Penicillin-Streptomycin-Amphotericin B Solution (ATCC PCS-999-002) Phenol Red (ATCC PCS-999-001)
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    Structured Review

    ATCC mesenchymal stem cell basal medium
    Functional Effects of HTLV-1 EVs on Angiogenesis and Inflammation. a An angiogenesis assay in technical and biological triplicate was used to determine the effect of distinct HTLV-1 EVs (2 k, 10 k, and 100 k) on tubular formation in a co-culture of <t>mesenchymal</t> <t>stem</t> cells (MSCs) and aortic endothelial cells (AEC). EV-treated cells (1:2000 recipient <t>cell</t> to EV ratio). Positive control cells received complete, undiluted <t>medium.</t> Additional dose of EV treatment was given to the cells at day 4. Representative images were taken at days 3 and 6 showing tubular formation in response to the indicated treatment. b The image processing software WIMASIS was used to calculate the percentage of area covered by tubules (n = 3) on day 3 and day 6. c RT-qPCR results showing env RNA copy numbers of mesenchymal stem cells treated with CEM EVs (control) and different populations of HTLV-1 EVs: 2 k (left panel), 10 k (middle panel), and 100 k (right panel). A set of dashed black vertical lines (---) were used to indicate baseline env RNA copy numbers. A set of dashed red vertical lines ( ) were used to indicate the levels of starting material, suggestive of the minimum env RNA copy numbers necessary for EVs to increase vial spread in MSc. d Western blot analysis for core histones (H3, H2A, H2B, and H4), linker histone (H1) and actin in HUT102 EVs (2 k, 10 k, and 100 k). e GAPDH DNA levels (representative of nucleosomes) in 2 k, 10 k, and 100 k HUT102 EVs treated with proteinase K and DNase/RNase were evaluated by was quantitated by q-PCR. A two-tailed student t-test was used to evaluate statistical significance with “**” for p-values ≤ 0.01, indicating the level of significance relative to untreated (Control) samples
    Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs is a sterile phenol red free liquid tissue culture medium intended for use as one component in a complete ATCC Primary Cell Solutions system This low serum 2 FBS system is designed to support mesenchymal stem cells derived from various normal human tissues including lipoaspirates and umbilical cord Mesenchymal Stem Cell Basal Medium contains essential and non essential amino acids vitamins other organic compounds trace minerals and inorganic salts To support the proliferation and plating efficiency of various types of adult stem cells Mesenchymal Stem Cell Basal Medium must be supplemented with the appropriate cell specific growth kit When using this complete media system the growth of undifferentiated mesenchymal stem cells is supported without the use of feeder layers extracellular matrix proteins or other substrates For mesenchymal stem cells derived from human lipoaspirates e g Adipose Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 011 or umbilical cord Umbilical Cord Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 010 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Adipose and Umbilical derived MSCs Low serum ATCC PCS 500 040 For mesenchymal stem cells derived from bone marrow Bone Marrow derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 012 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Bone Marrow derived MSCs ATCC PCS 500 041 Optional media supplements Gentamicin Amphotericin B Solution ATCC PCS 999 025 Penicillin Streptomycin Amphotericin B Solution ATCC PCS 999 002 Phenol Red ATCC PCS 999 001
    https://www.bioz.com/result/mesenchymal stem cell basal medium/product/ATCC
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mesenchymal stem cell basal medium - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "Extracellular vesicles from HTLV-1 infected cells modulate target cells and viral spread"

    Article Title: Extracellular vesicles from HTLV-1 infected cells modulate target cells and viral spread

    Journal: Retrovirology

    doi: 10.1186/s12977-021-00550-8

    Functional Effects of HTLV-1 EVs on Angiogenesis and Inflammation. a An angiogenesis assay in technical and biological triplicate was used to determine the effect of distinct HTLV-1 EVs (2 k, 10 k, and 100 k) on tubular formation in a co-culture of mesenchymal stem cells (MSCs) and aortic endothelial cells (AEC). EV-treated cells (1:2000 recipient cell to EV ratio). Positive control cells received complete, undiluted medium. Additional dose of EV treatment was given to the cells at day 4. Representative images were taken at days 3 and 6 showing tubular formation in response to the indicated treatment. b The image processing software WIMASIS was used to calculate the percentage of area covered by tubules (n = 3) on day 3 and day 6. c RT-qPCR results showing env RNA copy numbers of mesenchymal stem cells treated with CEM EVs (control) and different populations of HTLV-1 EVs: 2 k (left panel), 10 k (middle panel), and 100 k (right panel). A set of dashed black vertical lines (---) were used to indicate baseline env RNA copy numbers. A set of dashed red vertical lines ( ) were used to indicate the levels of starting material, suggestive of the minimum env RNA copy numbers necessary for EVs to increase vial spread in MSc. d Western blot analysis for core histones (H3, H2A, H2B, and H4), linker histone (H1) and actin in HUT102 EVs (2 k, 10 k, and 100 k). e GAPDH DNA levels (representative of nucleosomes) in 2 k, 10 k, and 100 k HUT102 EVs treated with proteinase K and DNase/RNase were evaluated by was quantitated by q-PCR. A two-tailed student t-test was used to evaluate statistical significance with “**” for p-values ≤ 0.01, indicating the level of significance relative to untreated (Control) samples
    Figure Legend Snippet: Functional Effects of HTLV-1 EVs on Angiogenesis and Inflammation. a An angiogenesis assay in technical and biological triplicate was used to determine the effect of distinct HTLV-1 EVs (2 k, 10 k, and 100 k) on tubular formation in a co-culture of mesenchymal stem cells (MSCs) and aortic endothelial cells (AEC). EV-treated cells (1:2000 recipient cell to EV ratio). Positive control cells received complete, undiluted medium. Additional dose of EV treatment was given to the cells at day 4. Representative images were taken at days 3 and 6 showing tubular formation in response to the indicated treatment. b The image processing software WIMASIS was used to calculate the percentage of area covered by tubules (n = 3) on day 3 and day 6. c RT-qPCR results showing env RNA copy numbers of mesenchymal stem cells treated with CEM EVs (control) and different populations of HTLV-1 EVs: 2 k (left panel), 10 k (middle panel), and 100 k (right panel). A set of dashed black vertical lines (---) were used to indicate baseline env RNA copy numbers. A set of dashed red vertical lines ( ) were used to indicate the levels of starting material, suggestive of the minimum env RNA copy numbers necessary for EVs to increase vial spread in MSc. d Western blot analysis for core histones (H3, H2A, H2B, and H4), linker histone (H1) and actin in HUT102 EVs (2 k, 10 k, and 100 k). e GAPDH DNA levels (representative of nucleosomes) in 2 k, 10 k, and 100 k HUT102 EVs treated with proteinase K and DNase/RNase were evaluated by was quantitated by q-PCR. A two-tailed student t-test was used to evaluate statistical significance with “**” for p-values ≤ 0.01, indicating the level of significance relative to untreated (Control) samples

    Techniques Used: Functional Assay, Angiogenesis Assay, Co-Culture Assay, Positive Control, Software, Quantitative RT-PCR, Western Blot, Polymerase Chain Reaction, Two Tailed Test

    2) Product Images from "Concentrated Growth Factors (CGF) Induce Osteogenic Differentiation in Human Bone Marrow Stem Cells"

    Article Title: Concentrated Growth Factors (CGF) Induce Osteogenic Differentiation in Human Bone Marrow Stem Cells

    Journal: Biology

    doi: 10.3390/biology9110370

    Monostrate of human Bone Marrow Stem Cells (hBMSC) in the presence of Concentrated growth factors (CGF). Immediately after its preparation CGF was placed directly on monolayer hBMSC culture, in Mesenchymal Stem Cell (MSC) Basal medium, for the indicated times.
    Figure Legend Snippet: Monostrate of human Bone Marrow Stem Cells (hBMSC) in the presence of Concentrated growth factors (CGF). Immediately after its preparation CGF was placed directly on monolayer hBMSC culture, in Mesenchymal Stem Cell (MSC) Basal medium, for the indicated times.

    Techniques Used:

    Related Articles

    Activity Assay:

    Article Title: Performance-enhanced mesenchymal stem cells via intracellular delivery of steroids
    Article Snippet: .. Metabolic activity, viability, and morphology assays Metabolic activity of MSCs was assessed by XTT (ATCC) following 24, 48, or 72 hour exposure to budesonide. ..

    Article Title: Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential
    Article Snippet: On day 2 and 5 after seeding, the number of EC tube formations were counted at 10× magnification by inverted microscopy and reported as n° of tube structures/field. .. Evaluation of anti-inflammatory activity of AD-MSCs and NHDFs The anti-inflammatory activity of AD-MSCs and HNDFs was tested by applying AD-MSCs-CM and HNDFs-CM on the U937 monocyte cell line (ATCC Manassas VA, USA), both in the absence and the presence of inflammatory stimuli LPS 1 μg/ml (Sigma) and TNFα 25 ng/ml (Sigma). .. The expression of the adhesion molecules CD54, CD44, CD62L and CD49d on U937 were analyzed by FC.

    Migration:

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow
    Article Snippet: .. Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in . .. Representative images of tubular structures formed by HUVEC incubated with mbMSC and bmMSC conditioned medium, for 48 and 72 h of culture, carried out in normoxia and 1% hypoxia, are shown in A–H.

    Expressing:

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow
    Article Snippet: .. Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in . .. Representative images of tubular structures formed by HUVEC incubated with mbMSC and bmMSC conditioned medium, for 48 and 72 h of culture, carried out in normoxia and 1% hypoxia, are shown in A–H.

    Functional Assay:

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow
    Article Snippet: .. Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in . .. Representative images of tubular structures formed by HUVEC incubated with mbMSC and bmMSC conditioned medium, for 48 and 72 h of culture, carried out in normoxia and 1% hypoxia, are shown in A–H.

    In Vitro:

    Article Title: A Regulatory miRNA–mRNA Network Is Associated with Tissue Repair Induced by Mesenchymal Stromal Cells in Acute Kidney Injury
    Article Snippet: A FACSCanto II flow cytometer (BD, Beckton Dickson) was used for cell acquisition, and the FlowJo software was used for data analysis. .. Coculture of MSCs or MVs with Renal Tubular Cells For in vitro assays, approximately 2 × 105 of renal epithelial tubular cells (MM55.K, ATCC® CRL-6436TM) were seeded in six-well plates (TPP, USA) and treated with nephrotoxic drug cisplatin (8 µg/mL) for 48 h, and two additional treated groups with cisplatin were co-cultured in contact with MSCs (v/v 1:1, 1 × 105 ) or MVs (50 µg/ml, sequentially each 6 h) for 48 h (Figure S4C in Supplementary Material). .. Subsequently, cells were trypsinized and subjected to analysis for apoptosis, cell proliferation, and oxidative stress analysis using the respective kits: Alexa Fluor® 488 annexin V/Dead Cell Apoptosis kit, CellTrace™ Violet Cell Proliferation kit, and MitoSOX™ Red Mitochondrial Superoxide Indicator kit (Life Technologies, USA), following the manufacturer’s recommendations (n = 6).

    Cell Culture:

    Article Title: Mesenchymal Stem/Stromal Cells in Stromal Evolution and Cancer Progression
    Article Snippet: .. Supplementary Material Cell culture: Human bone marrow mesenchymal stromal cells (MSCs) and HEK 293T cells (ATCC, Manassas, VA, USA) were cultured in α -MEM 10% FBS α -MEM powder (Invitrogen cat#12000-014, Paisley, UK), defined “FBS” (HyClone cat#SH3007003M, Logan, UT, USA) supplemented with penicillin-streptomycin (Sigma, St Louis, MO, USA), glutamine (Invitrogen cat#25030024), and minimal essential amino acids, NEAA (Invitrogen cat#11140035). .. The α -MEM powder was dissolved to commercial cell culture level water (Sigma).

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion
    Article Snippet: Cell lines and sphere cultures Unless otherwise stated, all reagents were purchased from Sigma-Aldrich. .. HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium). ..

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Mesenchymal Stem/Stromal Cells in Stromal Evolution and Cancer Progression
    Article Snippet: .. Supplementary Material Cell culture: Human bone marrow mesenchymal stromal cells (MSCs) and HEK 293T cells (ATCC, Manassas, VA, USA) were cultured in α -MEM 10% FBS α -MEM powder (Invitrogen cat#12000-014, Paisley, UK), defined “FBS” (HyClone cat#SH3007003M, Logan, UT, USA) supplemented with penicillin-streptomycin (Sigma, St Louis, MO, USA), glutamine (Invitrogen cat#25030024), and minimal essential amino acids, NEAA (Invitrogen cat#11140035). .. The α -MEM powder was dissolved to commercial cell culture level water (Sigma).

    other:

    Article Title: Characterization of the antimicrobial activity of the cell-penetrating peptide TAT-RasGAP317-326
    Article Snippet: Pseudomonas aeruginosa strain PA14 was grown either in LB or BM2 medium.

    Stem Cell Culture:

    Article Title: Circadian Rhythm and Cartilage Extracellular Matrix Genes in Osseointegration: A Genome-Wide Screening of Implant Failure by Vitamin D Deficiency
    Article Snippet: For Col10a1, the primer/probe cocktail was customly designed based on ref. XM_001053056.1 (Applied Biosystems, Carlsbad, CA). .. Bone marrow mesenchymal stem cell culture on titanium disks and vitamin D supplementation Mouse bone marrow derived mesenchymal stem cells (D1 ORL UVA [D1], ATCC® Number: CRL-s12424™) were plated at a density of 16,000 cells/well either on 48-well culture dishes (Becton Dickinson Labware, Franklin Lakes, NJ) or on sterile 10-mm Ti discs with surface of dual acid-etching and discrete crystalline deposition of HA nanoparticles (Biomet3I) . .. D1 cells were maintained with minimum Dulbecco's Modified Eagle's medium (1X DMEM, Cellgro, Mediatech Inc., Manassas, VA) supplemented with 10% fetal bovine serum (FBS, Benchmark, Gemini Bio-Products, West Sacramento, CA), and 1% penicilin-streptomicin (PS, MD Biomedicals, Thermo Fisher Scientific) under a humidified atmosphere of 5% CO2 in air, 37°C.

    Derivative Assay:

    Article Title: Circadian Rhythm and Cartilage Extracellular Matrix Genes in Osseointegration: A Genome-Wide Screening of Implant Failure by Vitamin D Deficiency
    Article Snippet: For Col10a1, the primer/probe cocktail was customly designed based on ref. XM_001053056.1 (Applied Biosystems, Carlsbad, CA). .. Bone marrow mesenchymal stem cell culture on titanium disks and vitamin D supplementation Mouse bone marrow derived mesenchymal stem cells (D1 ORL UVA [D1], ATCC® Number: CRL-s12424™) were plated at a density of 16,000 cells/well either on 48-well culture dishes (Becton Dickinson Labware, Franklin Lakes, NJ) or on sterile 10-mm Ti discs with surface of dual acid-etching and discrete crystalline deposition of HA nanoparticles (Biomet3I) . .. D1 cells were maintained with minimum Dulbecco's Modified Eagle's medium (1X DMEM, Cellgro, Mediatech Inc., Manassas, VA) supplemented with 10% fetal bovine serum (FBS, Benchmark, Gemini Bio-Products, West Sacramento, CA), and 1% penicilin-streptomicin (PS, MD Biomedicals, Thermo Fisher Scientific) under a humidified atmosphere of 5% CO2 in air, 37°C.

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    ATCC bone marrow msc
    <t>MG63</t> OS cells form spheres and their number is increased by the presence of <t>MSC.</t> (A) Homotypic cultures of CSC and their co-culture with MSC for 6 days were photographed at a magnification of 4x, representative images (scale bar 500 μm, black arrows show the spheres with higher size); (B) spheres were counted and expressed as sphere forming efficiency (number of spheres formed / number of cells seeded × 100) (*p
    Bone Marrow Msc, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MG63 OS cells form spheres and their number is increased by the presence of MSC. (A) Homotypic cultures of CSC and their co-culture with MSC for 6 days were photographed at a magnification of 4x, representative images (scale bar 500 μm, black arrows show the spheres with higher size); (B) spheres were counted and expressed as sphere forming efficiency (number of spheres formed / number of cells seeded × 100) (*p

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: MG63 OS cells form spheres and their number is increased by the presence of MSC. (A) Homotypic cultures of CSC and their co-culture with MSC for 6 days were photographed at a magnification of 4x, representative images (scale bar 500 μm, black arrows show the spheres with higher size); (B) spheres were counted and expressed as sphere forming efficiency (number of spheres formed / number of cells seeded × 100) (*p

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques: Co-Culture Assay

    MSC secrete pro-inflammatory cytokines when exposed to OS CSC whereas CSC are unaffected. (A) Scheme of the experiments used to evaluate the secreted proteins showed in the following panels; (B) TGF- β1 and IL-6 ELISA assays of the supernatants collected from the different chambers of the transwell with MSC co-cultured with HOS-CSC. TGFβ1 is secreted exclusively by MSC co-coltured in the presence of stem-like cells whereaseIL-6 is secreated by MSC independently from the co-cultuing with MSC. However secreated IL-6 levels were increased in MSC when co-cultured with CSC. The amount of IL-6 detected in lanes 2 and 4 is most likely due to leakage from the upper chamber, as no amount could be detected in the control in lane 6, where no MSC were seeded; (C) IL-6 ELISA assay for the evaluation of the same phenomenon with MG63-CSC secretion. The trend ofMG63-CSC was the same observed for HOS-CSC cells.

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: MSC secrete pro-inflammatory cytokines when exposed to OS CSC whereas CSC are unaffected. (A) Scheme of the experiments used to evaluate the secreted proteins showed in the following panels; (B) TGF- β1 and IL-6 ELISA assays of the supernatants collected from the different chambers of the transwell with MSC co-cultured with HOS-CSC. TGFβ1 is secreted exclusively by MSC co-coltured in the presence of stem-like cells whereaseIL-6 is secreated by MSC independently from the co-cultuing with MSC. However secreated IL-6 levels were increased in MSC when co-cultured with CSC. The amount of IL-6 detected in lanes 2 and 4 is most likely due to leakage from the upper chamber, as no amount could be detected in the control in lane 6, where no MSC were seeded; (C) IL-6 ELISA assay for the evaluation of the same phenomenon with MG63-CSC secretion. The trend ofMG63-CSC was the same observed for HOS-CSC cells.

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture

    Model for the circuit between MSC and HOS-CSC. Recruitment of MSC to the tumor environment leads to enhanced proliferation of OS stem cells. The presence of CSC, in turn, leads to a consistent secretion of TGFβ1 that activates a stromal autocrine loop that might be responsible for the activation of NF-kB genes and IL-6 secretion by MSC. Indeed, neutralization of TGFβ1 reduces the amount of secreted IL-6. Pro-tumorigenic effects of MSC, via IL-6, including induction of HOS-CSC migration and sphere growth, can be counteracted also by IL-6 neutralizing antibody. The presence of MSC is also responsible for increased expression of adhesion molecules involved in intra- or extra-vasation and the expression of MET can be counteracted by IL-6 neutralization.

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: Model for the circuit between MSC and HOS-CSC. Recruitment of MSC to the tumor environment leads to enhanced proliferation of OS stem cells. The presence of CSC, in turn, leads to a consistent secretion of TGFβ1 that activates a stromal autocrine loop that might be responsible for the activation of NF-kB genes and IL-6 secretion by MSC. Indeed, neutralization of TGFβ1 reduces the amount of secreted IL-6. Pro-tumorigenic effects of MSC, via IL-6, including induction of HOS-CSC migration and sphere growth, can be counteracted also by IL-6 neutralizing antibody. The presence of MSC is also responsible for increased expression of adhesion molecules involved in intra- or extra-vasation and the expression of MET can be counteracted by IL-6 neutralization.

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques: Activation Assay, Neutralization, Migration, Expressing

    The presence of HOS-CSC enhance the secretion of TGFβ1 from MSC that, in turn. autocrinally induce the activation of inflammatory pathways. (A) Scheme of the experiments used to evaluate the secreted proteins showed in the following panels. The secreted protein (only the supernatant in the MSC upper compartment was) (B) and mRNA expression (C) of TGFβ1 in MSC when co-cultured with HOS-CSC, after 3 days (*p

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: The presence of HOS-CSC enhance the secretion of TGFβ1 from MSC that, in turn. autocrinally induce the activation of inflammatory pathways. (A) Scheme of the experiments used to evaluate the secreted proteins showed in the following panels. The secreted protein (only the supernatant in the MSC upper compartment was) (B) and mRNA expression (C) of TGFβ1 in MSC when co-cultured with HOS-CSC, after 3 days (*p

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques: Activation Assay, Expressing, Cell Culture

    Stromal cells enhance HOS-CSC migration via IL-6 and the expression of adhesion molecules. (A) We assessed whether treatment with Tocilizumab affected HOS-CSC migration. MSC were treated with Tocilizumab [100 μg/mL] 2 hours prior CSC seeding. HOS spheres were trypsinized and single cells were let to migrate for 3 hours. As a control, medium only was added in the lower chambers, representative images; (B) Quantification of the migration assay shown in panel (A) (*p

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: Stromal cells enhance HOS-CSC migration via IL-6 and the expression of adhesion molecules. (A) We assessed whether treatment with Tocilizumab affected HOS-CSC migration. MSC were treated with Tocilizumab [100 μg/mL] 2 hours prior CSC seeding. HOS spheres were trypsinized and single cells were let to migrate for 3 hours. As a control, medium only was added in the lower chambers, representative images; (B) Quantification of the migration assay shown in panel (A) (*p

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques: Migration, Expressing

    TGFβ1-dependent IL-6 secretion is responsible for increased CSC sphere formation. (A) HOS-CSC were let to grow for 6 days in the presence of MSC; the latter cells were added every 24 hours with 100 μg/mL of Tocilizumab. Spheres were then photographed, counted, and quantified (scale bar, 500 μm). Representative figures and schematic representation of the performed assay; (B) Treatment with Tocilizumab significantly decreased the number of HOS-CSC spheres showed in panel A (*p

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: TGFβ1-dependent IL-6 secretion is responsible for increased CSC sphere formation. (A) HOS-CSC were let to grow for 6 days in the presence of MSC; the latter cells were added every 24 hours with 100 μg/mL of Tocilizumab. Spheres were then photographed, counted, and quantified (scale bar, 500 μm). Representative figures and schematic representation of the performed assay; (B) Treatment with Tocilizumab significantly decreased the number of HOS-CSC spheres showed in panel A (*p

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques:

    Gene expression of stem-cell markers confirmed the enhancement of stem-like features of CSC when are co-cultured with MSC in transwell. (A) Scheme of the co-culture system of HOS-CSCwith MSC used in this study. The co-culturing was prolonged for 3 days; (B) Sox2, Oct4, Nanog, and CXCR4 expression evaluated by Real Time PCR in HOS-CSC spheres that were cultured alone or with MSC (in transwell). Gene expression of CSC was also compared to parental HOS adherent cells (T0) (*p

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: Gene expression of stem-cell markers confirmed the enhancement of stem-like features of CSC when are co-cultured with MSC in transwell. (A) Scheme of the co-culture system of HOS-CSCwith MSC used in this study. The co-culturing was prolonged for 3 days; (B) Sox2, Oct4, Nanog, and CXCR4 expression evaluated by Real Time PCR in HOS-CSC spheres that were cultured alone or with MSC (in transwell). Gene expression of CSC was also compared to parental HOS adherent cells (T0) (*p

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques: Expressing, Cell Culture, Co-Culture Assay, Real-time Polymerase Chain Reaction

    MSC increase the proliferation of HOS-CSC. Homotypic cultures of CSC or co-cultures of CSC+MSC for 3 days were trypsinized, fixed and immunostained with anti-Ki67 antibody (green). Hoecst 33342(blue) was used to counterstain nuclei. (A) Representative image; (B) The percentage of Ki67 positive nuclei was quantified and expressed as ratio to the total number of cells (*p

    Journal: PLoS ONE

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion

    doi: 10.1371/journal.pone.0166500

    Figure Lengend Snippet: MSC increase the proliferation of HOS-CSC. Homotypic cultures of CSC or co-cultures of CSC+MSC for 3 days were trypsinized, fixed and immunostained with anti-Ki67 antibody (green). Hoecst 33342(blue) was used to counterstain nuclei. (A) Representative image; (B) The percentage of Ki67 positive nuclei was quantified and expressed as ratio to the total number of cells (*p

    Article Snippet: HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium).

    Techniques:

    T-shaped implant placed in vitamin D sufficient (V+) and deficient (V-) rats. A . T-shaped experimental implant with a hollow inner chamber was fabricated with Ti6Al4V and the implant surface was treated with dual acid-etching and discrete deposition of HA nanoparticles. T-shaped implant was placed in the osteotomy site of the distal end of rat femur. B . Serum chemistry evaluations of 25-hydroxy vitamin D3 (25D), parathyroid hormone (PTH), calcium (Ca), and phosphorous (P). *: p

    Journal: PLoS ONE

    Article Title: Circadian Rhythm and Cartilage Extracellular Matrix Genes in Osseointegration: A Genome-Wide Screening of Implant Failure by Vitamin D Deficiency

    doi: 10.1371/journal.pone.0015848

    Figure Lengend Snippet: T-shaped implant placed in vitamin D sufficient (V+) and deficient (V-) rats. A . T-shaped experimental implant with a hollow inner chamber was fabricated with Ti6Al4V and the implant surface was treated with dual acid-etching and discrete deposition of HA nanoparticles. T-shaped implant was placed in the osteotomy site of the distal end of rat femur. B . Serum chemistry evaluations of 25-hydroxy vitamin D3 (25D), parathyroid hormone (PTH), calcium (Ca), and phosphorous (P). *: p

    Article Snippet: Bone marrow mesenchymal stem cell culture on titanium disks and vitamin D supplementation Mouse bone marrow derived mesenchymal stem cells (D1 ORL UVA [D1], ATCC® Number: CRL-s12424™) were plated at a density of 16,000 cells/well either on 48-well culture dishes (Becton Dickinson Labware, Franklin Lakes, NJ) or on sterile 10-mm Ti discs with surface of dual acid-etching and discrete crystalline deposition of HA nanoparticles (Biomet3I) .

    Techniques:

    Expression of bone and cartilage extracellular matrix (ECM). A . Compiled microarray data of bone and cartilage ECM genes. Cartilage ECM genes were more significantly affected by implant placement and vitamin D deficiency than bone ECM genes. B . Microarray data for bone and cartilage ECM genes were confirmed by real time reverse transcription polymerase chain reaction (RTPCT).

    Journal: PLoS ONE

    Article Title: Circadian Rhythm and Cartilage Extracellular Matrix Genes in Osseointegration: A Genome-Wide Screening of Implant Failure by Vitamin D Deficiency

    doi: 10.1371/journal.pone.0015848

    Figure Lengend Snippet: Expression of bone and cartilage extracellular matrix (ECM). A . Compiled microarray data of bone and cartilage ECM genes. Cartilage ECM genes were more significantly affected by implant placement and vitamin D deficiency than bone ECM genes. B . Microarray data for bone and cartilage ECM genes were confirmed by real time reverse transcription polymerase chain reaction (RTPCT).

    Article Snippet: Bone marrow mesenchymal stem cell culture on titanium disks and vitamin D supplementation Mouse bone marrow derived mesenchymal stem cells (D1 ORL UVA [D1], ATCC® Number: CRL-s12424™) were plated at a density of 16,000 cells/well either on 48-well culture dishes (Becton Dickinson Labware, Franklin Lakes, NJ) or on sterile 10-mm Ti discs with surface of dual acid-etching and discrete crystalline deposition of HA nanoparticles (Biomet3I) .

    Techniques: Expressing, Microarray, Reverse Transcription Polymerase Chain Reaction

    Endothelial tube-like formation stimulated by MSC conditioned medium. Representative phase contrast microscopy images of tubular structures formed by HUVEC EA.Hy926 incubated with ( A , B , E , F ) mbMSC and ( C , D , G , H ) bmMSC conditioned medium derived of ( A – D ) 48 and ( E – H ) 72 h of culture, carried out in ( A , C , E , G ) normoxia and ( B , D , F , H ) 1% hypoxia, 20 h after plating in Matrigel TM GFR with serum-free MSC conditioned media. ( I , J ) Quantification of the total length and ( K , L ) branches number of the tubular network formed by HUVEC cultivated with the 48 and 72 h-CM of mbMSC and bmMSC. Values are expressed as mean ± SD (EGM2 48 h, n = 4; EGM-2 72 h, n = 5; mbMSC and bmMSC-CM, n = 5 for each condition); * p

    Journal: International Journal of Molecular Sciences

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow

    doi: 10.3390/ijms21249563

    Figure Lengend Snippet: Endothelial tube-like formation stimulated by MSC conditioned medium. Representative phase contrast microscopy images of tubular structures formed by HUVEC EA.Hy926 incubated with ( A , B , E , F ) mbMSC and ( C , D , G , H ) bmMSC conditioned medium derived of ( A – D ) 48 and ( E – H ) 72 h of culture, carried out in ( A , C , E , G ) normoxia and ( B , D , F , H ) 1% hypoxia, 20 h after plating in Matrigel TM GFR with serum-free MSC conditioned media. ( I , J ) Quantification of the total length and ( K , L ) branches number of the tubular network formed by HUVEC cultivated with the 48 and 72 h-CM of mbMSC and bmMSC. Values are expressed as mean ± SD (EGM2 48 h, n = 4; EGM-2 72 h, n = 5; mbMSC and bmMSC-CM, n = 5 for each condition); * p

    Article Snippet: Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in .

    Techniques: Microscopy, Incubation, Derivative Assay

    Impact of MSC conditioned medium over HUVEC EA.Hy926 migration by scratch wound assay in vitro. HUVEC migration curves in the presence of serum-free mbMSC and bmMSC-CM, obtained after ( A ) 48 and ( B ) 72-h of culture in normoxia and 1% hypoxia. Sample size for each condition are indicated. ( C , D ) Area under the curve (AUC) relative to HUVEC migration curves in the presence of 48 and 72 h-CM of mbMSC and bmMSC. ( E , F ) Wound recovery promoted by 48 and 72 h-CM of mbMSC and bmMSC at the point of 48 h after scratch. Values are expressed as mean ± SD; * p

    Journal: International Journal of Molecular Sciences

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow

    doi: 10.3390/ijms21249563

    Figure Lengend Snippet: Impact of MSC conditioned medium over HUVEC EA.Hy926 migration by scratch wound assay in vitro. HUVEC migration curves in the presence of serum-free mbMSC and bmMSC-CM, obtained after ( A ) 48 and ( B ) 72-h of culture in normoxia and 1% hypoxia. Sample size for each condition are indicated. ( C , D ) Area under the curve (AUC) relative to HUVEC migration curves in the presence of 48 and 72 h-CM of mbMSC and bmMSC. ( E , F ) Wound recovery promoted by 48 and 72 h-CM of mbMSC and bmMSC at the point of 48 h after scratch. Values are expressed as mean ± SD; * p

    Article Snippet: Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in .

    Techniques: Migration, Scratch Wound Assay Assay, In Vitro

    E. coli MG1655 and P. aeruginosa PA14 sensitivity to TAT-RasGAP 317-326 varies depending on the growth medium used. E. coli MG1655 was grown overnight in the indicated medium, diluted to 0.1 OD 600 and grown for 1h. Culture was then diluted 20 times and 10 µl was added per well of a 96-well plate containing serial dilutions of TAT-RasGAP 317-326 . OD 600 measurements after 16 hours of incubation in presence of the indicated concentrations of TAT-RasGAP 317-326 are shown and MIC is defined as the lowest concentration of TAT-RasGAP 317-326 that completely inhibits bacterial proliferation. IC 50 is defined as the concentration required to inhibit 50 % of growth and was calculated using GraphPad Prism 8. Indicated strains were grown overnight and diluted respectively in LB (A-B) , BM2 with 2mM MgSO 4 (Mg low ) (C-D) or BM2 with 20 µM MgSO 4 (Mg high ) (E-F) .

    Journal: bioRxiv

    Article Title: Characterization of the antimicrobial activity of the cell-penetrating peptide TAT-RasGAP317-326

    doi: 10.1101/2020.10.02.321802

    Figure Lengend Snippet: E. coli MG1655 and P. aeruginosa PA14 sensitivity to TAT-RasGAP 317-326 varies depending on the growth medium used. E. coli MG1655 was grown overnight in the indicated medium, diluted to 0.1 OD 600 and grown for 1h. Culture was then diluted 20 times and 10 µl was added per well of a 96-well plate containing serial dilutions of TAT-RasGAP 317-326 . OD 600 measurements after 16 hours of incubation in presence of the indicated concentrations of TAT-RasGAP 317-326 are shown and MIC is defined as the lowest concentration of TAT-RasGAP 317-326 that completely inhibits bacterial proliferation. IC 50 is defined as the concentration required to inhibit 50 % of growth and was calculated using GraphPad Prism 8. Indicated strains were grown overnight and diluted respectively in LB (A-B) , BM2 with 2mM MgSO 4 (Mg low ) (C-D) or BM2 with 20 µM MgSO 4 (Mg high ) (E-F) .

    Article Snippet: Pseudomonas aeruginosa strain PA14 was grown either in LB or BM2 medium.

    Techniques: Incubation, Concentration Assay

    TAT-RasGAP 317-326 is bactericidal against E. coli and P. aeruginosa . A-D) Overnight cultures of E. coli MG1655 in LB (A-B) and P. aeruginosa PA14 in BM2 Mg low (C-D) were diluted to 0.1 OD 600 . Peptide was added at the indicated concentrations 1 hour after dilution. Samples were taken at the indicated time points, serially diluted 10-fold in fresh LB and plated on LB agar plates. Number of colony forming unit per ml (CFU/ml) of original culture was calculated. E) E. coli MG1655 strain was incubated for one hour with or without 10 μM FITC-labelled TAT-RasGAP 317-326 . The bacterial sample was then labelled with 5 μg/ml FM4-64 and fixed with 4% paraformaldehyde solution. Incubation with DAPI was subsequently performed. Pictures were taken with a Zeiss LSM710 confocal microscope and analyzed using ImageJ software. F) E. coli bacteria treated as in (E) were fixed using glutaraldehyde and prepared for electron microscopy as described in Material and Methods section. Samples were imaged using transmission electron microscopy. Images were analyzed using ImageJ software.

    Journal: bioRxiv

    Article Title: Characterization of the antimicrobial activity of the cell-penetrating peptide TAT-RasGAP317-326

    doi: 10.1101/2020.10.02.321802

    Figure Lengend Snippet: TAT-RasGAP 317-326 is bactericidal against E. coli and P. aeruginosa . A-D) Overnight cultures of E. coli MG1655 in LB (A-B) and P. aeruginosa PA14 in BM2 Mg low (C-D) were diluted to 0.1 OD 600 . Peptide was added at the indicated concentrations 1 hour after dilution. Samples were taken at the indicated time points, serially diluted 10-fold in fresh LB and plated on LB agar plates. Number of colony forming unit per ml (CFU/ml) of original culture was calculated. E) E. coli MG1655 strain was incubated for one hour with or without 10 μM FITC-labelled TAT-RasGAP 317-326 . The bacterial sample was then labelled with 5 μg/ml FM4-64 and fixed with 4% paraformaldehyde solution. Incubation with DAPI was subsequently performed. Pictures were taken with a Zeiss LSM710 confocal microscope and analyzed using ImageJ software. F) E. coli bacteria treated as in (E) were fixed using glutaraldehyde and prepared for electron microscopy as described in Material and Methods section. Samples were imaged using transmission electron microscopy. Images were analyzed using ImageJ software.

    Article Snippet: Pseudomonas aeruginosa strain PA14 was grown either in LB or BM2 medium.

    Techniques: Incubation, Microscopy, Software, Electron Microscopy, Transmission Assay