成年大鼠骨骼肌成肌细胞的培养和鉴定

Culture and identification of adult rat skeletal myoblasts

目的探索成年大鼠骨骼肌成肌细胞的高浓度培养方法。方法以成年同种系Wistar大鼠为研究对象,采用两步消化法获取大鼠骨骼肌卫星细胞,进行体外培养,对获得的细胞进行形态学研究,以免疫组织化学方法进行鉴定。结果细胞增殖旺盛,分化良好,可融合成肌管。免疫细胞化学染色显示,骨骼肌卫星细胞呈弱阳性,肌管呈强阳性。结论两步消化法体外培养的骨骼肌卫星细胞具有良好的增殖与分化能力,用此种方法可培养出高纯度的骨骼肌成肌细胞,操作简单、污染少。

Objective To establish an experimental method for primary culture of adult rat skeletal myoblasts and to explore the biological characteristics of the cultured cells. Methods Rat skeletal muscle satellite cells were obtained with the two-step digestion using type I collagenase and trypsin and passaged following primary culture in vitro. The cells were counted and observed by light microscopy combined with immunocytochemical identification. Results Immunocytochemical analysis showed that most of the cells were stained positive for desmin and myosin, indicated factors for skeletal myocytes. Conclusion The modified method can produce high purified skeletal myoblasts with simple process.