鼠骨髓基质干细胞的特性及向成骨细胞的分化

目的分离纯化成年小鼠骨髓基质干细胞并进行定向诱导。方法利用Percoll液分离成年小鼠骨髓基质干细胞,Ter119、CD45磁珠纯化细胞,培养2-3代的细胞用矿化液进行定向诱导。对诱导前后的细胞进行免疫组化染色。结果 镜下见原代细胞呈多种形态。Ter119、CD45磁珠筛选可获得纯度达57.95％以上骨髓基质干细胞。碱性磷酸酶、Von Kossa染色阳性,油红染色弱阳性,矿化液诱导2周后,可形成钙结节。结论利用Percoll液结合磁珠分离法是离体条件下获得骨髓基质干细胞的一个简便有效的方法,纯化后定向诱导可得到较纯的成骨细胞。

The Characterization of Mice Bone Marrow Mesenchymal Stem Cells with Respect to Osteoblastic Differentiation

Objective To get purified bone marrow messenchymal stem cells (BMMSCs) and induce them into osteoblasts. Methods Bone marrow stromal cells (BMSCs) were selected with Percoll (1. 077g/cm3) and CD45 - Ter119 - BMMSCs were purified. BMMCs were stained with HE, Oil Red, PAS, Von Kossa, ALP, and Alizarin Reds. BMMSCs of passage 2-3 were cultured with mineralized medium to be induced into osteoblasts. Results BMSCs showed variouse shapes. Over 57. 95% of CD45-Terll9-BMSCs were BMMSCs with spindle shape. BMMSCs were weakly positively stained with Alizarin Red S, ALP, Von Kossa, and negatively stained with Oil Red. After cultured 14 days in mineralized medium, calcium knurs were observed by Alizarin Red S staining. Conclusion It was simple and efficient to get purified BMMSCs by Percoll and magnetic beads. Most of the BMMSCs became osteoblasts after specific induction.