Journal: ESC Heart Failure

Article Title: Metformin inhibits proliferation of residing fibroadipogenic progenitor cells from failing human hearts

doi: 10.1093/eschf/xvag001

Figure Lengend Snippet: CD45 − CD31 − CD34 + cells isolated from the human myocardium possess fibro- and adipogenic differentiation potential. For subsequent experiments, mononuclear cells were isolated from the left ventricle of human hearts from heart transplantation recipients. CD45 − CD31 − CD34 + cells were further selected by fluorescence-activated cell sorting for in vitro experimentation. ( A ) PDGFRA is a marker of mesenchymal stem cells (multipotent stem cells) and was exclusively expressed in one cluster with a fibroblast phenotype. CD34 was expressed in the same cluster as PDGFRα , as well as in a large cluster of PECAM1 (CD31) positive cells, which represent endothelial cells. PTPRC (CD45) was exclusively expressed in two clusters representing macrophages and T/NK cells. ( B ) Live CD45 − CD31 − CD34 + cells isolated by fluorescence-activated cell sorting. ( C ) Isolated CD45 − CD31 − CD34 + cells can be grown in cell culture and express PDGFRα assessed by immunofluorescence staining (magnification 10X). ( D and E ) Isolated CD45 − CD31 − CD34 + cells increases COL-1 expression when grown in fibrogenic media, suggesting that they differentiate into fibroblasts. ( F and G ) Isolated CD45 − CD31 − CD34 + cells start to express PLIN-1 when grown in adipogenic media, suggesting that they differentiate into adipocytes. Thus, these cells may represent human cFAPs. The image magnifiation for D-G is 20X

Article Snippet: Anti CD45 , Miltenyi , Cat. no. 130-114-123.

Techniques: Isolation, Transplantation Assay, Fluorescence, FACS, In Vitro, Marker, Cell Culture, Immunofluorescence, Staining, Expressing

Journal: ESC Heart Failure

Article Title: Metformin inhibits proliferation of residing fibroadipogenic progenitor cells from failing human hearts

doi: 10.1093/eschf/xvag001

Figure Lengend Snippet: Metformin in pharmacological doses (0.1 mM) inhibits proliferation of fibrogenic progenitors. Mononuclear non-muscle cells were isolated from the left ventricle of explanted human hearts from heart transplantation recipients. Subsequently, CD45 − CD31 − CD34 + cells were isolated by fluorescence-activated cell sorting for in vitro experimentation. ( A and B ) COL-1 and COL-6 mRNA expression in cFAPs grown in fibrogenic media and incubated with metformin (0.1 mM) and cimetidine (0.1 mM) alone and in combination. ( C ) Quantification of proliferation assay as per cent 5-ethynyl-2'-deoxyuridine (EDU) positive cells. Individual donors ( A–C ) are indicated by the shape of the dots. ( D ) Cell proliferation assays (EDU) and apoptosis assays (TUNEL) performed on cFAPs in cell culture incubated with metformin (0.1 mM) and cimetidine (0.1 mM) alone and in combination (magnification 10X) (* P < .05)

Article Snippet: Anti CD45 , Miltenyi , Cat. no. 130-114-123.

Techniques: Isolation, Transplantation Assay, Fluorescence, FACS, In Vitro, Expressing, Incubation, Proliferation Assay, TUNEL Assay, Cell Culture