骨髓间质干细胞贴壁筛选法的提纯

Purification of the adherence culture for the Mesenchymal stem cells

目的体外分离贴壁筛选大鼠骨髓间质干细胞（mesenchymaIstemcells，MSCs），并对MSCs的表面标志进行测定。方法分离4周龄的大鼠股骨、胫骨骨髓进行体外贴壁培养、筛选，取第1～10代的MSCs观察形态，并用流式细胞仪检测其表面标志。结果大鼠MSCs细胞形态呈长梭形，培养筛选至第3—6代时流式细胞术检测：CD29、CD90阳性率均在90％以上，CIM5阳性率在10％以下。结论本方法分离纯化出的是MSCs，第3～6代时纯度最高。

Objective To isolate and select rat bone marrow mesenchymal stem cells （MSCs） in vitro by means of adherence culture, and to determine the surface marker of MSCs. Methods The bone marrow from femur and tibia of four- weeks-old rat was isolated, purified and cultured in vitro. The morpholog of MSCs from generations 1 to 10 were observed by microscope constantly and the surface marker was analyzed by flow cytometry. Results The MSCs were elongated. The cells of third to sixth generations were analyzed by flow cytometry ： the positive rate of CD29 and CD90 were all more than 90% and CD45 was less than 10%. Conclusions This method could isolate and purify MSCs. The third-sixth generations have the highest purity.