大鼠骨髓间充质干细胞的体外培养与鉴定

In-vitro culture and identification of rat bone marrow mesenchymal stem cells

目的:探讨大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)的体外培养方法及其生物学特性。方法:体外分离、培养大鼠BMSCs,进行细胞形态学观察、生长曲线测定、细胞表面标记物检测;并采用体外诱导分化方法对BMSCs多向分化潜能进行鉴定。结果:大鼠BMSCs传代后细胞生长状态相对稳定,呈梭形;流式细胞仪鉴定CD90阳性率为88.2%,CD29阳性率为98%,CD34阴性率为2.8%,CD45阴性率为2.7%。成骨诱导21 d后,茜素红染色后可在显微镜下观察到矿化结节形成。成脂诱导14 d,油红O染色后显微镜下可见红色脂滴形成。结论:BMSCs在体外培养中贴壁生长,增殖较快;表达骨髓组织来源干细胞的表面标记物CD29、CD90;诱导后能向成骨细胞、脂肪细胞分化。

Objective： To culture and indentify rat bone marrow mesenchymal stem cells（BMSCs）.Methods： BMSCs were separated from SD rats bone marrow and cultured in vitro.Morphological characteristics were observed and the cell surface antigens were detected by flow cytometry;the multi-directional differentiation potency were examined by inductive differentiation experiments.Cell proliferation was detected with MTT assay.Results： The subcultured BMSCs grew well and most BMSCs presented shuttle shape under phase contrast microscope.Flow cytometry examination showed that CD29 and CD90 expressed positively and CD34,CD45 negatively in the second passage of BMSCs.After adipogenic induction for 14 days,lipid droplets in BMSCs were observed by oil red staining.After osteogenetic induction for 21 days,calcific nodus were found.Conclusion： Rat BMSCs can be cultured in vitro and maintain proliferation and differentiation potential.