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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。
OBJECTIVE: To establish an in vitro culture system of bone marrow mesenchymal stem cells (MSCs), and to induce osteogenic differentiation to investigate the in vitro culture methods and differentiation conditions of osteoblasts. Methods: MSCs were isolated from adult SD rats by whole bone marrow adherent culture. After induced by conditioned medium, morphological changes were observed and cell growth curve, immunocytochemical detection and alkaline phosphatase activity were performed. The formation of the determination. Results: The SD rat MSCs were isolated and cultured in vitro with long spindle shape and S-shaped cell growth curve. Under the induction conditions, the cell alkaline phosphatase activity was significantly increased, and the emergence of mineralized nodules. CONCLUSION: SD rat MSCs can be successfully isolated and cultured in vitro. The cultured cells can differentiate into osteoblasts and can be used as seed cells for bone tissue engineering.