摘要
目的探讨成年大鼠原代骨骼肌卫星细胞体外培养方法,观察细胞的增殖、成肌分化特性。方法采用改进的两步酶消化法结合差速贴壁技术,获得较高纯度的骨骼肌卫星细胞。免疫组化鉴定所获取细胞的肌源性标志desmin。MTT法观察骨骼肌卫星细胞的体外增殖特性并绘制生长曲线图。同时观察不同培养条件下骨骼肌卫星细胞的成肌分化特性。结果所分离的原代骨骼肌卫星细胞纯度在90%以上。细胞体外培养时在第3天进入对数生长期,5～6d进入增殖平台期。细胞体外培养时无须特殊诱导可以自发出现成肌定向分化,相互融合形成具有自发收缩特性的肌管细胞。结论体外培养获取的骨骼肌卫星细胞数量可以满足组织工程研究的需要。
Objective To discuss isolation and culture methods of primary rat skeletal muscle satellite cells and observe its characterization of proliferation and differentiation in vitro culture. Methods Primary muscle satellite cells of relative high purity were obtained by improved two-steps enzymatic digestion combined with pre-plating techniques. Marker of muscle-derived cell, desmin, was identified by histocytochemistry. The characterization of cell proliferation was studied by the way of MTT assay experiment. Differentiation of muscle satellite cell under different culture conditions in vitro was also studied. Results The purity of harvested muscle satellite cell was more than 90 percent. When cultured in vitro, the latent phase of the cell is at the Ist to 2nd days, platform phase is at the 5th to 6th day. It shows strong proliferative ability in vitro culture and could fuse with each other into myotube cells without special induction. Conclusions Muscle satellite cells by in vitro culture were enough to meet study of tissue engineering and cell transplantation.
出处
《实验动物与比较医学》
CAS
2007年第1期20-24,共5页
Laboratory Animal and Comparative Medicine
基金
上海市科委资助项目(044119605)
关键词
骨骼肌卫星细胞
增殖
分化
鉴定
Skeletal muscle satellite cell
Proliferation
Differentiation
Identification
