壁虎骨骼肌卫星细胞的原代培养与鉴定

Primary cultivation and identification of satellite cells of Gekko japonicus skeletal muscles

目的:建立一种有效的壁虎骨骼肌卫星细胞分离、培养及鉴定体系。方法:取壁虎后肢肌,采用I型胶原酶消化法获得细胞悬液,经差速贴壁法纯化骨骼肌卫星细胞,通过Pax7免疫细胞化学方法鉴定骨骼肌卫星细胞。用此法也分离到小鼠骨骼肌卫星细胞。结果:(1)分离到的细胞呈小圆形,培养2天后可见纺锤形细胞。(2)免疫细胞化学染色显示,分离得到的壁虎骨骼肌卫星细胞表达Pax7。Pax7阳性的小鼠骨骼肌卫星细胞达99%以上。培养3天后卫星细胞开始增殖。结论:成功建立壁虎骨骼肌卫星细胞的纯化和鉴定方法,为进一步研究卫星细胞在高等哺乳动物损伤修复中的可塑性奠定基础。

Objective： To establish a method for isolation,culture and identification of satellite cells from gecko skeletal muscles.Methods： Cell suspension was obtained from posterior limb skeletal muscles of gecko by collagenase I digestion,and the muscle satellite cells were purified by the method of differential adherence.Finally satellite cells were identified with monoclonal antibody against Pax7.Satellite cells of mice skeletal muscles were obtained in accordance with this method as well.Results：（1）The cells isolated by this method showed high survival rate and strong proliferate ability.（2）Immunocytochemical staining showed that the obtained cells of gecko express Pax7.Pax7-positive satellite cells of mice was more than 99%.Conclusion： The method for satellite cell isolation and purification from gecko was successfully established.