小鼠骨骼肌卫星细胞的分离培养和鉴定

摘要

采用胶原酶和胰酶联用的酶消化法分离小鼠骨骼肌卫星细胞，应用差速贴壁法进行纯化，并利用RT-PCR和免疫荧光染色方法对分化前后细胞的标志基因进行鉴定。结果显示：分离出的肌卫星细胞生长状态良好，RT-PCR和免疫荧光染色显示肌卫星细胞Pax7和MyoD呈阳性表达，诱导分化形成肌管后，分化标志基因MyoG和MHC呈阳性表达。本研究成功从小鼠肌肉组织中分离出了肌卫星细胞，并具有很好的体外分化能力，可以为肌肉的发育分化和损伤修复研究提供良好的细胞模型。%In this study, mouse skeletal muscle satellite cells were isolated by collagenase and trypsinase digestion method. Then the satellite cells were purified and induced to differentiate into myotubes, and the marker genes of the cells before or after the differentiation were detected by RT-PCR and immunocytochemistry technology. The results show that the isolated muscle satellite cells were healthy, and expressed the marker genes of paired protein box(Pax7) and myogenic differentiation antigen (MyoD). After inducing to the myotubes, myogenin(MyoG) and myosin heavy chain(MHC)showed positive expression. The results indicate that mouse skeletal muscle satellite cells of high purity, which can provide cell sources for muscle development and damage repair study, are successfully isolated.