小鼠骨髓间质干细胞的体外培养与多向分化潜能的鉴定

In Vitro Culture and Multipotential Differentiation Identification of Murine Bone Marrow Mesenchymal Stem Cells

【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 develop a method of culture, purification, enrichment, and identification of murine bone marrow mesenchymal stem cells in vitro, and identify the muhipotential differentiation under different induction media. [Methods] The bone marrow cells were collected from 3-18 months old male C57BL/6J mice （48 mice）, and they were isolated, enriched and expanded by using bone marrow adherent culture. The cells were purified by using immunomagnetic microbeads, and the growth curve of MSCs from passages 3, passages 10, and passages 20 were protracted. The cell morphology, as well as their osteogenic differentiation, adipogenic differentiation, and myogenic differentiation under different induction media were examined. [Results] Osteoblast, adipocyte, and myoblast could be induced from murine bone marrow mesenchymal stem cells in vitro. The MSC retained their morphology, properties and multipotential differentiation after more than 30 passages in culture as well as from frozen stocks. [Conclusion] The MSC can be isolated, expanded and enriched according to our method, and the cells have stable properties, muhipotential differentiation.