primary human adipose derived mscs  (PromoCell)

Journal: International Journal of Molecular Sciences

Article Title: Human Omental Mesothelial Cells Impart an Immunomodulatory Landscape Impeding B- and T-Cell Activation

doi: 10.3390/ijms23115924

Figure Lengend Snippet: Phenotypic characterization of human omental mesothelial cells (OMC) and human mesenchymal stem cells (MSC). ( A ) Phase contrast pictures of confluent OMC and MSC cultures displaying typical cobblestone-type and fibroblastic morphologies, respectively. Scale bar is 100 µm. ( B ) Flow cytometric analysis of OMC and MSC for expression of hematopoietic (CD45), endothelial (CD31), HLA class II (HLA-DR/DP/DQ), and stromal/mesenchymal (CD90, CD29, CD44, CD73, CD73, CD13, CD105, and CD166) cell markers. Histograms for OMC and MSC are red and blue, respectively. M1 bar marks positivity delimited from isotype histograms (not shown). ( C ) Immunofluorescence analysis of OMC and MSC showing highly expressed epithelial (pan-cytokeratin or pan-CK) and mesothelial (Wilm’s tumor protein 1 or WT1) cell markers in OMC, but not in MSC, while the cell–cell junction protein β-catenin was expressed in both cell types. ( D ) Multilineage differentiation assay of OMC and MSC revealed limited adipogenic differentiation of OMC (Oilred O staining) compared to MSC. In contrast, OMC displayed more similar osteogenic and chondrogenic differentiation compared to MSC. ( A – C ) OMC used are ALIC1 cells.

Article Snippet: Primary human mesenchymal stem cells (MSC) derived from subcutaneous adipose tissue acquired by liposuction (ATCC ® PCS-500-011) were used as a reference of cells with immunosuppressive properties [ , ].

Techniques: Expressing, Immunofluorescence, Wilms Tumor Assay, Differentiation Assay, Staining

Journal: Stem Cell Research & Therapy

Article Title: Hypoxic preconditioning potentiates the trophic effects of mesenchymal stem cells on co-cultured human primary hepatocytes

doi: 10.1186/s13287-015-0218-7

Figure Lengend Snippet: HPc potentiates the hepatotrophic and anti-apoptotic effects of co-culture with MSCs. a Representative histogram of DCFDA to measure intracellular ROS fluorescence intensity of serum-deprived, HPc-, and HPc + NAC-pretreated MSCs. b Bar chart of normalised MFI of intra-MSC ROS activity. Values are mean ± standard deviation ( n = 6). * p < 0.05 and ** p < 0.01, versus NPc-MSCs. c Albumin secretion and d urea synthesis of mono- and co-cultured hepatocytes with NPc- or HPc-MSCs, with or without NAC 10 mM. e Soluble CCK18 and f CK18 releases of mono- and co-cultured hepatocytes. g Staurosporine cytotoxicity assay of mono-cultured hepatocytes and resistance of co-cultured hepatocytes to 1 μM staurosporine-induced cellular apoptosis and h total death. Values are mean ± standard deviation ( n = 6). ** p < 0.01, versus hepatocyte mono-culture; ## p <0.01, versus co-culture with NPc-MSCs; + p <0.05 and ++ p <0.01, versus co-culture with HPc-MSCs; ^^ p <0.01, versus hepatocyte mono-culture treated with 1 μM staurosporine. CCK18 Caspase-cleaved cytokeratin 18, CK18 Cytokeratin 18, Co Co-culture; DCFDA Dichloro-dihydrofluorescein diacetate acetyl ester, HPc Hypoxia-preconditioned, Hx Hepatocyte mono-culture; MFI Median fluorescence intensity, NAC N-acetylcysteine, NPc Normoxia-preconditioned, SD Serum-deprived, SF Serum-free , SS Staurosporine

Article Snippet: Human primary adipose tissue-derived MSCs were purchased from Invitrogen Ltd (Paisley, UK), and passages 6−8 (P6–P8) of MSCs were subcultured using a low-serum MSC expansion medium (Invitrogen).

Techniques: Co-Culture Assay, Fluorescence, Activity Assay, Standard Deviation, Cell Culture, Cytotoxicity Assay

Journal: Stem Cell Research & Therapy

Article Title: Hypoxic preconditioning potentiates the trophic effects of mesenchymal stem cells on co-cultured human primary hepatocytes

doi: 10.1186/s13287-015-0218-7

Figure Lengend Snippet: Decreased TNF-α and increased TGF-β1 in hepatocytes co-cultured with MSCs related to apoptosis. a TNF-α secretion of mono-/co-cultured hepatocytes and NPc-/HPc-MSCs. b TGF-β1 secretion of mono-/co-cultured hepatocytes and NPc-/HPc-MSCs. c Neutralisation with anti-TNF inhibits cellular apoptosis and, to a lesser extent, total death of mono-cultured hepatocytes, switching the main cell death mode from apoptosis to necrosis. d Neutralisation of MSCs with anti-TGF-β1 induces cellular apoptosis and, to a lesser extent, total death of co-cultured hepatocytes, switching the main cell death mode from necrosis to apoptosis. Values are mean ± standard deviation ( n = 6). * p < 0.05 and ** p <0.01, versus control mono- or co-culture; ^^ p <0.01, versus co-culture with HPc-MSCs; ## p <0.01, versus control mono-culture; ++ p <0.01, versus co-culture. α-TGF-β1 Anti-transforming growth factor-β1, α-TNF Anti-tumour necrosis factor, CCK18 Caspase-cleaved cytokeratin 18, CK18 Cytokeratin 18, Co Co-culture, HPc Hypoxia-preconditioned, Hx Hepatocyte mono-culture, ind. Indirect, Mono mono-culture, MSC Mesenchymal stem cell, NAC N-acetylcysteine, NPc Normoxia-preconditioned, UD undetectable

Article Snippet: Human primary adipose tissue-derived MSCs were purchased from Invitrogen Ltd (Paisley, UK), and passages 6−8 (P6–P8) of MSCs were subcultured using a low-serum MSC expansion medium (Invitrogen).

Techniques: Cell Culture, Standard Deviation, Co-Culture Assay

Journal: Stem Cell Research & Therapy

Article Title: Hypoxic preconditioning potentiates the trophic effects of mesenchymal stem cells on co-cultured human primary hepatocytes

doi: 10.1186/s13287-015-0218-7

Figure Lengend Snippet: Dependence of co-culture hepatotrophic and HPc-induced potentiative effects on ROS-dependent MSC deposition of extracellular collagen. a Extracellular and b cellular collagen contents of mono-/co-cultured hepatocytes and NPc-/HPc-MSCs with or without NAC addition. c Inhibitory effect of MaIBA on extracellular collagen deposit of NPc- and HPc-MSCs. d Effects of 5 mM MaIBA pretreatment on main cellular death mode of hepatocytes co-cultured with NPc- versus HPc-MSCs. e and f Picro-sirius red staining (200×) for cellular and extracellular collagen of MSCs in mono-culture ( left panel ) and co-culture ( right panel ); nuclei are counterstained with hematoxylin. Values are mean ± standard deviation ( n = 6). * p <0.05 and ** p <0.01, versus control mono- or co-culture; ^^ p <0.01, versus co-culture with HPc-MSCs; ## p <0.01, versus control mono-culture; ++ p <0.01, versus co-culture. CCK18 Caspase-cleaved cytokeratin 18, CK18 Cytokeratin 18, Co Co-culture, HPc Hypoxia-preconditioned, Hx Hepatocyte mono-culture, ind. Indirect, MaIBA N-(methylamino)-isobutyric acid, Mono mono-culture, MSC Mesenchymal stem cell, NAC N-acetylcysteine, NPc Normoxia-preconditioned, PSR OD Picro-sirius red OD reading , UD undetectable

Article Snippet: Human primary adipose tissue-derived MSCs were purchased from Invitrogen Ltd (Paisley, UK), and passages 6−8 (P6–P8) of MSCs were subcultured using a low-serum MSC expansion medium (Invitrogen).

Techniques: Co-Culture Assay, Cell Culture, Staining, Standard Deviation