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目的采用组织块培养技术探索大鼠骨骼肌卫星细胞的原代培养方法。方法以成年SPF级Sprague-Dawley大鼠为研究对象,采用组织块培养法获取大鼠骨骼肌卫星细胞,并与C2C12成肌细胞进行比较,对两种细胞进行形态学研究及采用免疫荧光和免疫组织化学法测定两种细胞α-actin蛋白和Desmin蛋白的表达及分布,从而对骨骼肌卫星细胞进行鉴定。结果通过组织块培养法获取的细胞增殖旺盛,分化良好。免疫细胞荧光和免疫组织化学实验结果显示,α-actin蛋白和Desmin蛋白在两种细胞胞浆中均有分布。结论用组织块培养法获取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卫星细胞。
Objective To explore the primary culture method of rat skeletal muscle satellite cells by tissue culture technique. Methods Sprague-Dawley adult Sprague-Dawley rats were used as experimental materials. Skeletal muscle satellite cells of rat were obtained by tissue culture method and compared with C2C12 myoblasts. Morphological study of the two kinds of cells and immunofluorescence and immunofluorescence Histochemical methods were used to determine the expression and distribution of α-actin and Desmin proteins in skeletal muscle satellite cells. Results The cells obtained by tissue culture method proliferated well and differentiated well. Immunofluorescence and immunohistochemistry results showed that α-actin protein and Desmin protein were distributed in both cytoplasm. Conclusion Skeletal muscle satellite cells obtained by tissue culture method have good proliferation and differentiation ability, and high purity skeletal muscle satellite cells can be cultured by this method.