人骨髓间充质干细胞体外培养及生物学特性观察

Biologic characteristics of human bone marrow-derived mesenchymal stem cells cultured in vitro

目的:观察体外培养的人骨髓间充质干细胞(MSCs)部分生物学特性。方法:自健康人骨髓中分离出MSCs,进行原代培养,第5d首次更换培养液,以去除未贴壁细胞,以后每周换液2次,细胞铺满整个培养瓶底90%时开始传代培养。倒置相差显微镜下观察原代及传代细胞形态及生长特性,并绘制传代细胞的生长曲线。流式细胞仪检测第3代MSCs细胞表面分子CD29、CD44,以进行MSCs鉴定。结果:MSCs体外培养为贴壁生长。MSCs原代培养约18～21d才达传代要求;传代培养细胞生长潜伏期仅24～36h,对数增长期约3～5d,之后为平台期,4～6d传1代。传至第8代,出现凋亡征象。第3代MSCs细胞表面CD29、CD44的表达率分别为90.6%和91.5%,MSCs均一性较高。结论:从人骨髓中分离出的MSCs在体外环境中具有很强的自我更新和分裂增殖能力。

Aim: To investigate the biological characteristics of human bone marrow-derived mesenchymal stem cells (MSCs) in vitro. Methods: Human MSCs were isolated from bone marrow and purified by centrifuge and cultured in vitro. The proliferation and growth characteristics of MSCs were observed in primary and passage culture. Results: Human bone marrow-derived MSCs showed active proliferation capacity in vitro in primary and passage cultures.Conclusion: MSCs derived from human bone marrow have been successfully cultured. It founds a base for further investigation and using of mesenchyaml stem cells.