Modified culture conditions of bone marrow mesenchymal stem cells and directional differentiation

LIU Rong; HE Yong-jiang; GUO Lv-hua; YANG Xue-chao; CHEN Jun

Abstract：Objective To establish simple, rapid and effective separation and purification methods for bone mar-row mesenchymal stem cells ( BMSCs) in vitro.Methods The BMSCs were isolated by whole bone marrow centrifugal adherent method, and cultured in allogeneic serum medium ( allogeneic serum group) , fetal bovine serum medium ( fetal bovine serum group) or serum free medium (serum free group), respectively.First medium change was performed 12, 24, 48, 72 hours or 5 days after culturing.P3 BMSCs growth curve was drawn.Expression of CD44 and vimentin on BM-SCs, and CD31 and vWF on vascular endothelium like cells were identified by immunofluorescence.Expression of calcium nodules were detected by alizarin red.Results (1) There was no significant difference in cell morphology and growth rate between allogeneic serum group and fetal bovine serum group.(2) The cell growth rates of allogeneic serum group and fe-tal bovine serum group were higher than that of serum free group.(3) The CD44 and vimentin immunofluorescence stai-ning of allogeneic serum group and fetal bovine serum group were higher than that of serum free group.(4) Alizarin red staining showed a large number of orange red mineralized nodule formation after osteogenic induction for 14 days.(5) En-dothelial cell surface markers CD31 and vWF were positive.Conclusion The improved culture conditions for bone marrow cells harvests with phenotypic and functional characteristics of BMSCs.