人骨髓间充质干细胞的分离培养、多向分化与鉴定

Objective　To establish a method for effective isolation and multiplication of human bone marrow derived mesenchymal stem cells and verification by morphology and multi-potential differentiation. Methods MSCs were isolated from human bone marrow by combination of gradient centrifugation and different adherent time method. Morphology and growth characteristics were examined by phase contrast microscopy. Cell surface markers CD13, CD29, HLA-2, CD34, CD45 and HLA-DR were tested by flow cytometer. MSCs were induced in vitro respectively to osteoblasts by β-glycerophosphate, L-ascorbicacid-2-phosphate, dexamethasone and FK506; to chondrocytes by TGF-beta 3, recombulin, sodium pyruvate and selenious acid; to adipocytes by IBMX and indomethacin; to endothelial cells by VEGF and bFGF. Results Cells were spindle-shaped and presented active proliferation in primary and passage cultures. The Cell surface markers CD13, CD29, HLA-2 were positive and CD34, CD45 and HLA-DR were negative. Cells were successfully induced into osteoblasts, chondrocytes, adipocytes and endothelial cells. Conclusion Method is established to isolate and multiply MSCs from human bone marrow effectively and to verify by morphology and its multi-potential differentiation.