哈萨克羊骨骼肌卫星细胞的分离培养及鉴定

Isolation, culture and identification of skeletal muscle satellite cells from Kazakh sheep

为了建立可稳定传代的哈萨克羊骨骼肌卫星细胞培养体系,以90日龄的哈萨克羊胎羊后腿肌为试验材料,采用酶消化法(胶原酶与胰蛋白酶)对绵羊骨骼肌卫星细胞进行分离,差速贴壁法对细胞纯化;对得到的骨骼肌卫星细胞进行形态学鉴定;使用Pax7特异性标志蛋白进行免疫荧光鉴定。经过免疫荧光染色的Pax7基因的免疫阳性产物,呈明显的绿荧光,分布于细胞质中。本研究成功从哈萨克胎羊中分离出了具有良好增殖分化能力且便于传代的骨骼肌卫星细胞,为研究调控哈萨克羊肌肉生长发育的分子机制,为完善哈萨克羊肉品质的研究提供基础。

This study was to establish a stable cell culture system for skeletal muscles of Kazakh sheep. The thigh muscles of 90-day-old Kazakh sheep were used as experimental materials, satellite cells were separated the muscles by enzyme digestion(with collagenase and trypsin), and then the cells were purified using the differential adherence method. Finally, morphological identification was performed of the obtained skeletal muscle satellite cells;and immunofluorescence identification was done on htem using the Pax7-specific marker protein. Immunofluorescent staining was carried out on the immune-positive products of the Pax7 gene which were clearly bright green and were distributed in the cytoplasm. The results indicated that skeletal muscle satellite cells with good capabilities of proliferation and differentiation ability and of easy passage were successfully isolated from Kazakh fetal sheep, which provided a basis for research on the molecular mechanism of muscle development and improving the mutton quality of Kazakh sheep.