大鼠骨髓基质干细胞体外分离、纯化、培养的实验研究

A Study on the Methods of Separation,Depuration and Cultivation in Vitro for Bone Marrow Stromal Cells of Rats

目的：探讨Wistar大鼠骨髓基质干细胞（ BMSCs）体外分离、纯化、培养的方法。方法对5周龄Wistar大鼠予1%戊巴比妥钠（0.006 ml/g）行腹腔麻醉,成功后于大鼠双侧后肢股骨、胫骨提取BMSCs,用贴壁培养法分离、纯化、培养BMSCs,观察细胞形态变化及绘制细胞生长曲线了解其生长、传代、扩增特性,并应用流式细胞仪检测细胞表面标志物行细胞鉴定。结果应用贴壁培养法分离、纯化、培养的大鼠BMSCs增殖迅速、生长曲线良好,经细胞表面标志物检测证实为BMSCs,且细胞具有很好的均一性（96.36%）。结论贴壁培养法能够很好地分离、纯化、培养大鼠BMSCs,并且获得的细胞具有很强的增殖能力,取第3~6代细胞用于实验研究最为合适。

Objective To explore the methods of separation, depuration and cultivation in vitro for bone marrow stromal cells （ BMSCs） of rats. Methods Five-week-old Wistar rats were anesthetized through abdominal cavities using 1% sodium pentobarbital （0. 006 ml/g）, and then BMSCs were extracted from the rats＆#39;femurs and tibias. The adherent culture method was used to separate, purify and culture the BMSCs. The changes in cell morphology were observed, and characteristics of cell growth, passage and amplification were studied by drawing curve of growth. The cells were identi-fied with flow cytometry. Results The BMSCs obtained with adherent culture method had rapid proliferation and good growth curve, which proved BMSCs by the detection of cell surface marker, and the cells had good homogeneity （96. 36%）. Conclusion Adherent culture can well separate, purify and culture the BMSCs, and the obtained cells have better proliferative capacity, and the 3th -6th generations are more suitable for experimental research.