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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 a method for the isolation and culture of human bone marrow-derived mesenchymal stem cells (hMSCs) in vitro and to investigate the biological characteristics of hBMSCs such as phenotype, cell cycle and growth curve. Methods The hBMSCs were isolated and cultured by density gradient centrifugation. The phenotypes of hBMSCs were detected by flow cytometry. The cell cycle and hBMSCs growth curve were drawn. Results The hBMSCs with high purity were separated by density gradient centrifugation. The expression of CD105, CD13, CD34, CD45 and HLA-DR in hBMSCs were detected by flow cytometry. The growth curve of hBMSCs was S-type. Cells are in a relatively inactive quiescent state. Conclusion The hBMSCs with high purity can be obtained by in vitro isolation and culture of bone marrow using density gradient centrifugation. The hBMSCs have unique biological characteristics.