人骨髓间充质干细胞体外培养及生物学特性的研究

Study of Isolation,Cultivation and Biological Identification of Human Bone Marrow Mesenchymal Stem Cells

目的建立体外分离培养人骨髓间充质干细胞(Human Bone-marrow Mesenchymal Stem Cells,hBMSCs)方法,并就hBMSCs表型鉴定、细胞周期、生长曲线等生物学特性进行初步研究。方法采用密度梯度离心法分离培养hBMSCs,流式细胞仪检测hBMSCs表型鉴定、细胞周期、绘制hBMSCs生长曲线。结果密度梯度离心法能分离出纯度较高的hBMSCs;流式细胞仪检测hBMSCs CD105、CD13阳性,CD34、CD45、HLA-DR表达阴性;hBMSCs的生长曲线呈S型;hBMSCs细胞生长周期提示大部分细胞处于相对不活跃的静止期。结论采用密度梯度离心法体外分离培养骨髓可获得纯度较高的hBMSCs,hBMSCs具有独特的生物学特征。

Objective To establish the method of isolation and cultivation of human bone marrow-mesenchymal stem cells （hBMSCs）, and to conduct the preliminary study on the specific surface markers, cell cycle and growth curve of hBMSOs. Methods Bone marrow aspirate were harvested and cultured with density- gradient centrifugation method： The third passage cells were used to detect the specific surface markers and cell cycle with Flow Cytometry. The fifth passage cells were cultured for another days to draw the growth curve. Results Higher purity hBMSCs can be achieved by the density - gradient centrifugation method. The cells were positive for CD105 and OD13,. but negative for OD34, OD45, and HLA-DR. The： growth curve of hBMSCs was ＂S＂ shape. （85.62 ± 1. 21） % of the hBMSCs were found at G0/G1 phase, that meant most of the hBMSCs were in the metabolic stage. Conclusions The method of density- gradient centrifugation is applicable to get higher purity hBMSCs, and the hBMSCs have the special biological characters.