ASCs were cultured at a density of 1 x 10 5 in T25 flasks and counted every 24 h for 6 days. ( B ) Comparison of the growth rate between WT and trappc9-deficient (KO) ASCs. Arrowheads point to cells containing two or more nuclei. ( A ) Confocal images showed that ASCs isolated from WT and trappc9-null (KO) mice expressed mesenchymal stem cell markers and were devoid of hematopoietic CD45. Both WT and KO ASCs at passage-3 were subjected to analyses. ASCs were isolated from WT and trappc9-deficient mice and cultured as in Methods. MSCs (-HSCs).Ĭompromised self-renewal of ASCs cultured from trappc9-null mice. ( E ) Real-time PCR analysis of mesenchymal lineage marker expression in MSCs following a 3-week treatment with mesenchymal differentiation media after co-culture with HSCs ( n = 5). After co-culture of MSCs and HSCs for 3, 5, and 7 days, cells were collected, sorted based on EGFP expression, and both cell types were detected using stem cell markers by flow cytometry.

MSCs were sorted on day -1, and we waited until they attached to the bottom of the dish on day 0. ( D ) MSC and HSC population in whole mesenchymal and hematopoietic cells. * p < 0.05 and ** p < 0.01 between direct and indirect co-culture. ( B, C ) Cell proliferation of MSCs ( B ) and HSCs ( C ) 2, 4, 6, and 8 days after co-culture ( n = 6). Microscopic views of MSCs (upper) and HSCs (bottom). ( A ) Representative flow cytometric profiles of MSCs (upper, CD31 -, CD45 -, Ter119 -, Sca-1 +, and PDGFR-alpha + cells) and HSCs (bottom, c-Kit +, Lin -, and Sca-1 + cells). Clonal assay of MSCs after co-culture with HSCs.

Super fm canli