摘要
目的 :探讨骨骼肌卫星细胞体外培养、鉴定的方法及其生物学特性。方法 :采用 型胶原酶和胰蛋白酶二步消化法获取大鼠骨骼肌卫星细胞 ,进行体外原代和传代培养。观察细胞的形态 ,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 :二步消化法适用于骨骼肌卫星细胞的获取。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。 α sarcometric actin和 m yosin免疫细胞化学染色 ,骨骼肌卫星细胞呈弱阳性 ,肌管呈强阳性。结论 :体外培养的骨骼肌卫星细胞具有良好的增殖与分化能力 。
Objective:To establish the methods for culture,identification and biological characteristics of skeletal muscle satellite cells.Methods:Rat skeletal muscle satellite cells were obtained by the twostep method of collegenaseⅠ and trypsin,and were subject to primary as well as secondary culture in vitro.Morphological observation,growth curve and rate of myotube formation (RMF) were observed to evaluate the proliferative and differentiation ability of skeletal muscle satellite cells.The cells were identified with cellular immunochemical stain.Results:The twostep method with collegenaseⅠ and trypsin was suitable for collection of skeletal muscle satellite cells.The cells showed strong proliferative ability in the proliferative media and could form myotubes in differentiation media.Skeletal muscle satellite cells were weak positive and myotubes were strong positive by cellular immunochemical stain with αsarcometric actin and myosin.Conclusions:Skeletal muscle satellite cells cultured in vitro have strong proliferative and differentiation ability and can be used to tissue engineering and gene therapy.
出处
《中国危重病急救医学》
CAS
CSCD
2002年第2期104-106,共3页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目 ( No.39870 811)
关键词
骨骼肌卫星细胞
鉴定
生物学特性
细胞培养
skeletal muscle satellite cells
culture
identification
biological characteristics
