SD大鼠骨髓间充质干细胞的体外分离培养鉴定及骨向诱导分化研究

目的：建立骨髓间充质干细胞（MSCs）体外分离培养体系,并进行骨向分化诱导以探讨大鼠MSCs的体外培养方法及向成骨细胞分化的条件。方法：采用全骨髓贴壁筛选培养法分离成年SD大鼠MSCs,经条件培养液诱导培养后,进行细胞形态学观察、绘制细胞生长曲线、免疫细胞化学检测及碱性磷酸酶活性、钙结节的形成测定。结果：SD大鼠MSCs在体外分离培养扩增形态呈长梭形,细胞生长曲线呈S形。诱导条件下,细胞碱性磷酸酶活性明显增高,并出现了矿化结节。结论：SD大鼠MSCs体外分离培养体系能够成功建立,培养出的细胞能向成骨细胞分化,可以作为骨组织工程的种子细胞。

Isolation, culture, identfication and induced differentination of bone marrow mesenchymal stem cells into osteoblasts in sd rats

Objective： To establish an isolation and culture system of SD rat bone marrow mesenchymal stem cells and osteogenic differentiation in vitro in order to study the condition for induced differentiation to osteoblasts. Method： The bone rMSCs were isolated from adult SD rats and purified by wall adhesion culture procedure and passaged in vitro. Examining the rMSCs under microscope, immunocytochemical staining and drawing cell growth curve. The passage cells were stained by alkaline phosphatase （ALP） and calcifying nodule Vonkossa. Result： Colonies of rMSCs were formed in the shuttle and the growth curve reports the S form through isolation, purification and amplification. In the cells of osteogenic differentiation, the ALP expression and calcifying nodule Vonkossa were positive. Conclusion： The isolation and culture system of rMSCs can be successfully established and the rMSCs sustained their osteogenic differentiation potential into vitro, rMSCs can be seed cells for tissue engineering of bone.