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【目的】建立小鼠骨髓间质干细胞(MSC)体外培养、纯化、扩增方法,并进行体外定向诱导条件下的多向分化潜能鉴定。【方法】取3～18月雄性C57BL/6J小鼠(共48只)骨髓细胞,贴壁培养后,用免疫磁珠法纯化,观察纯化后的细胞形态,对3、10、20代的MSC进行细胞生长曲线绘制;并检测细胞在不同诱导条件下向成骨细胞、脂肪细胞、肌肉细胞的分化能力。【结果】小鼠MSC在体外可以诱导成成骨细胞、脂肪细胞、肌肉细胞。MSC连续传代培养达30代以上,细胞的形态、抗原表达无改变,并保持多向分化潜能。【结论】该方法获得的MSC具有高度增殖、多向分化潜能、生物学特性稳定的特点。
【Objective】 To establish a method for culturing, purifying and amplifying mouse bone marrow mesenchymal stem cells (MSCs) in vitro and to identify the potential of multidrug differentiation under in vitro induction. 【Methods】 Bone marrow cells of male C57BL / 6J mice (a total of 48) were collected from March to August. After culturing them adherently, the cells were purified by immunomagnetic beads method. The cell morphology was observed. The cell growth curve was drawn and the differentiation ability of cells to osteoblasts, adipocytes and muscle cells under different induction conditions was tested. 【Results】 Mouse MSCs can be induced into osteoblasts, adipocytes and muscle cells in vitro. MSC continuous subculture up to 30 generations, the cell morphology, antigen expression was unchanged, and maintain the potential of multi-directional differentiation. 【Conclusion】 The MSCs obtained by this method have the characteristics of high proliferation, multi-directional differentiation and stable biological characteristics.