农业生物技术学报2011，Vol.19Issue(5)：856-863,8.DOI:10.3969/i.issn.1674-7968.2011.05.011
利用单根肌纤维法分离和培养猪骨骼肌卫星细胞及其成肌诱导分化
Isolation and Culture of Porcine Skeletal Muscle Satellite Cells by Single Muscle Fiber and Their Muscle Differentiation
摘要
Abstract
The present study established a porcine satellite cells ire vitro model by means of the single fiber method to clarify the proliferation and myogenesis. Satellite cells can dissociate from the myofibers which isolated from porcine extensor digitorum longus (EDL) after digesting by collagenase I . Then the cells were detected by their immunocytochemistry, growth curve, myogenesis induction and Western blot. The results showed that the cells adhered to the myofibers originally and then moved outside. More than 90 percent of the cells expressd the marker genes paired protein box(Pax7) and myogenic differentiation antigen(AfyoZ>). After myogenesis induction, satellite cells began to convergence ,showed orientation growth, and finally formed multinucleated myotubule with expression of myogenin and myosin heavy chain (MyHC). Moreover, MyoD protein was highly expressed in proliferated stage, while the protein of myogenin and MyHC was detected only in the terminal differentiated stage. This study successfully cultured porcine satellite cells by means of the single fiber method and obtained high purity skeletal muscle satellite cells which can provide cell source for viable transplantation and cure related diseases.关键词
骨骼肌纤维/卫星细胞/Pax7/猪Key words
Skeletal muscle single fiber/ Satellite cells/ Pax7/ Porcine引用本文复制引用
刘月光,史新娥,沈清武,袁媛,杨秋梅,高晓娟,陈宗正,杨公社..利用单根肌纤维法分离和培养猪骨骼肌卫星细胞及其成肌诱导分化[J].农业生物技术学报,2011,19(5):856-863,8.基金项目
本研究由国家转基因生物新品种培育科技重大专项基金(NO.2009zx08009-157B)资助 （NO.2009zx08009-157B）
