C57BL6小鼠骨髓间充质干细胞的分离培养鉴定

目的建立一种简单实用的小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)体外分离培养方法,为BMSCs的进一步研究和应用提供实验材料和依据。方法采用直接贴壁法和分阶段更换培养基的方法分离纯化BMSCs,显微镜下观察其形态,应用流式细胞技术鉴定细胞表面标记。结果显微镜下BMSCs贴壁生长后呈梭形,形态均一,经流式细胞仪检测第2代BMSCs高表达CD34、Sca-1,不表达CD45。结论本实验方法能简单高效的获得高纯度BMSCs,且细胞生物学特性稳定,适用于对BMSCs的进一步研究和应用。

Isolation,culture,identification of mesenchymal stem cells from C57BL/6 mouse bone marrow

Objective To establish a simple and practical method for isolation and culture of mesenchymal stem cells from C57BL/6 mouse bone marrow(BMSCs) in vitro,and to provide material and a experimental base for further study and application of BMSCs.Methods The BMSCs were isolated from C57BL/6 mouse bone marrow,isolated and purified by adherent culture combined with using different medium in the different stages of BMSCs.The morphology was observed by invert-microscopy and the membrane antigens of BMSCs were detected by flow cytometric analysis.Results The BMSCs were adherent cells with a homogeneous spindle-shaped morphology under invert-microscope.The second generation of BMSCs tested were negative for the hematopoietic cell marker CD45.However,they expressed high levels of CD34 and Sca-1.Conclusions In this study,a simple and efficient method to obtain high purity of BMSCs has been established,and BMSCs isolated and purified by this method have stable biological properties.So this method is suitable to be used in the further research and application of BMSCs.