大鼠肌肉卫星细胞的分离、鉴定与诱导分化  

Isolation,Identification and Differentiation of Rat Muscle Satellite Cell

作　　者：焦泽华[1] 魏著英[1] 白春玲[1] 段彪[1] 程磊[1] 李光鹏[1] 

机构地区：[1]内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室,呼和浩特010021

出　　处：《农业生物技术学报》2011年第2期302-307,共6页Journal of Agricultural Biotechnology

基　　金：国家高技术研究发展计划(No.2007AA100505;No.2008AA10Z159)资助

摘　　要：肌肉卫星细胞是组织特异性干细胞,负责着骨骼肌的生长和再生。尽管发现卫星细胞的存在已经有50多年的历史了,但分离鉴定肌肉卫星细胞的方法仍然不稳定。本研究使用两步酶消化和差速贴壁相结合的方法分离大鼠(Rattus norvegicus)肌肉卫星细胞。免疫细胞化学染色显示,分离的大鼠肌肉卫星细胞高表达Desmin;通过RT-PCR鉴定所分离的细胞表达肌源性干细胞因子Myf5及Myod1;成肌诱导48h后,卫星细胞开始形成多核的肌管;成脂诱导1周后,细胞内开始出现脂滴,2周后充脂细胞明显增多。研究结果提示,分离的卫星细胞具有肌源性,有发育形成肌管和向成脂细胞分化的能力。Satellite cells are regarded as a population of muscle-specific committed progenitors which are responsible for the postnatal maintenance,growth,repair and regeneration of skeletal muscles.Although satellite cells have been first isolated for 50 years,the isolation protocols are still to be modified.In this study,the combination of two-step enzyme digestion and purification with different speed adherence protocols were used to isolate rat（Rattus norvegicus） satellite cells.Analyses of the cells with desmin immunocytochemical staining indicated that the cells were derived from skeletal muscle;RT-PCR of Myf5 and Myod1 showed that they expressed muscle-specific transcription factor;Myotubes began to form at 48 h in induced-culture medium;Lipid droplets began to appear in cells after 1 week,and the number of adipose cells increased significantly after 2 weeks.The results suggested that isolated satellite cells could be induced into adipogenesis,and had the abilities to develop into myobubes and differentiated into adipose cells.

关 键 词：大鼠 肌肉卫星细胞 成肌诱导 成脂诱导 

分 类 号：S858.31[农业科学—临床兽医学]