成人骨髓间充质干细胞的体外培养及生物学性质

Culture and biological characteristics of adult human bone marrow-derived mesenchymal stem cells in vitro

目的 :建立人骨髓间充质干细胞 (BM- MSCs)分离培养方法 ,观察细胞形态 ,探讨 BM- MSCs生物学特征。方法 :从人骨髓中利用 Percoll分离有核细胞 ,培养于含 10 %胎牛血清的低糖 DMEM培养基中 ,台盼蓝拒染法测定细胞增殖状态。应用流式细胞仪测定细胞周期及细胞表面抗原特征。结果 ;BM- MSCs具有纤维样细胞形态特征 ,独特的表型 ,即 CD2 9、CD4 4、CD16 6和 HL A- ABC阳性 ,CD34、CD4 5和 HL A- DR阴性 ;细胞周期分析显示 :12 .8%处于 S期 ,76 .4 %处于 G0 / G1 期 ;细胞生长倍增时间为 2～ 3d。结论 :BM- MSCS是一群均一的单克隆细胞 ,具有独特的增殖特征及表面标记。

Objective To establish a method to isolate and culture the bone marrow-derived mesenchymal stem cells (BM-MSCs) in vitro and investigate their biologic characteristics. Methods The culture of mesenchymal stem cells was initiated from bone marrow low-density mononuclear cells separated by Pecoll centrifugation and maintained in low-glucose DMEM with 10% fetal calf serum. The cell growth pattern was evaluated by trypan blue exclusion. The cell cycle and surface antigenic features were analyzed by flow cytometry. Results The BM-MSCs were positive for CD29,CD44,CD166,HLA-ABC and negative for CD34, CD45,and HLA-DR. Cell cycle analysis showed that 12.8% of them was in S phase and 76.4% of them was in G 0/G 1 phase. The cells had a population doubling time of 2-3 days. Conclusion BM-MSCs are a homogenous monoclone population of cells with unique growth phenotype characteristics.