摘要
目的探讨体外培养的骨骼肌卫星细胞的生长与分化特性。方法取大鼠肱三头肌,采用二步消化法分离骨骼肌卫星细胞,体外进行原代和传代培养。通过生长曲线、细胞融合率研究骨骼肌卫星细胞的增殖与分化能力,利用免疫细胞化学染色对所获得的细胞进行鉴定。结果二步消化法适用于大鼠骨骼肌卫星细胞的分离,在生长培养基作用下,细胞增殖旺盛;在分化培养基条件下,细胞分化良好,可融合成肌管。骨骼肌特异性蛋白α-sarcometricactin和myosin免疫化学染色显示,骨骼肌卫星细胞弱阳性,肌管强阳性。结论体外培养的骨骼肌卫星细胞具有良好的生长与分化能力,适用于骨骼肌组织工程的构建与研究。
Objective To investigate the proliferation an d differentiation characteristics of skeletal muscle satellite cells in vitro.Methods Rat triceps brachii muscle was acqui red to separate skeletal muscle sate llite cells with the two-step method of collegen ase-Ⅰand trypsin.Cells were cultured and subcultured in vitro.Morphological observation,growth curve and rate o f myotube formation(RMF )were investigated to evaluate the pr oliferation and differentiation characteristics o f skeletal muscle satellite cells.T he cells were identified with immunochemical stain.Results The two-stpe method of collegenase-Ⅰand trypsin was fit to acquire skeletal muscle satellite cells.The cells showed strong proliferative a bility in the proliferative media an d could form myotubes in differentia tion media.Skeletal muscle satellite cells were faint positive,while myotu bes were strong positive with immunochemical stain ofα-sarcometric actin and myosin.Conclusions Skeletal muscle satellite cells cul tured in vitro have strong proliferation and differentiation ability and can be used in tissue-engineered skeletal muscle.[
基金
国家自然科学基金(No.39870811)
关键词
骨骼肌
卫星细胞
培养
生长
分化
Skeletal muscle Satellite cells Cul ture Proliferation Differentiation
