中文摘要：

目的分离培养鉴定小鼠的骨髓问充质干细胞，观察体外培养生长特性，并在特定条件下诱导分化，探讨其成脂成骨分化能力。方法采用全骨髓培养法培养小鼠骨髓间充质干细胞，观察干细胞的形态和生长特性，用流式细胞仪对其细胞表型CD29、CD90、CD34、CD44、CD31、CD45进行鉴定，成脂成骨诱导分化能力鉴定。结果原代分离的细胞培养24h，干细胞开始贴壁，胞体呈圆形，其他血细胞悬浮。培养3d，贴壁细胞逐渐变梭形；培养第4d，细胞开始分裂；随着细胞数目的增加，逐渐形成漩涡状排列，细胞形态呈梭形、三角形、多边形、不规则形；培养第10d，贴壁细胞长满瓶底的80％，按1：2传代。选择第3代骨髓干细胞分析显示，CD29、CD44、CD90阳性表达，CD31、CD34、CD45阴性表达。细胞成脂成骨诱导后染色鉴定呈阳性。结论采用全骨髓法培养可获得生长状态良好，增殖能力强的骨髓间充质干细胞，方法简便、实用。

英文摘要：

Objective To isolate,culture,and identify the mice~ bone marrow mesenchymal stem cells （BMSCs） ,and to observe the biological characteristics of those cells cultureed in vitro. To carry out the induced differentiation under certain conditions, and to discuss adipogenic and osteogenie differentiation capability. Methods Full bone marrow culture method was used to culture the mice BMSCs. Their cell morphous and growth characteristics were observed. Flow cytometry was used to identify the cell phenotypes of CD29, CD90, CD34, CD44, CD31, and CD45 and also the adipogenic and osteogenie differentiation ability. Results After the culture of primarily isolated cells for 24 h, the adherence of the stem cell occurred, and the cell body appeared the round shape. Other blood cells suspended. Three days after the culturing, the shape of the adherent cells changed into spindle shape. On d 4 after the culture, the cells began to divide. Mong with the increasing number of cells, whirl - like pattern gradually formed, andthe shapes of cells changed into shuttle, triangle, polygon, and irregular. On d 10, the attached cells covered 80% of the bottle bottom and they were passaged with the radio of 1 to 2. Analysis of the third passage BMSCs showed that the expressions of CD29, CD44, and CD90 were positive, but the expressions of CD31, CD34,and CIM5 were negative. The identification result by staining was positive after the adipogenic and osteogenie differentiation. Conclusion The full bone marrow culture method can obtain BMSCs with good growth status and strong proliferation ability. This method is simple and practical.