摘要
目的体外分离培养和纯化小鼠快、慢肌卫星细胞,观察细胞内β肌动蛋白(β-actin)的表达情况。方法采用胶原酶与胰蛋白酶联合消化法及差速贴壁法纯化ICR小鼠后肢骨骼肌腓肠肌(快肌)和比目鱼肌(慢肌)卫星细胞,免疫荧光抗体细胞化学染色法鉴定。取第2代快、慢肌卫星细胞提取总RNA,以GAPDH作为内参行RT-PCR,半定量分析快、慢肌卫星细胞内β-actin基因的相对表达量。结果体外分离及传代培养快、慢肌卫星细胞的快、慢肌肌浆蛋白表达阳性,细胞生长和增殖情况良好。快、慢肌卫星细胞内β-actin基因的相对表达量分别为3.71072和2.106028,两者比较差异有统计学意义(P<0.01)。结论小鼠快肌卫星细胞内β-actin表达高于慢肌,提示骨骼肌卫星细胞内β-actin表达可能与肌纤维类型有关。
Objective To culture and purify the mouse satellite cells of gastrocnemius and soleus muscles in vitro,and detect the expression of β-actin in cells.Methods Skeletal muscle satellite cells of gastrocnemius and soleus muscles of ICR mice were purified by collagenase and trypsin digestion and differential adhesion method,and were identified by immunofluorescent antibody cytochemical staining.Total RNA was extracted from the second generation of gastrocnemius and soleus muscles,and semiquantitative RT-PCR was ...
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2011年第1期26-30,共5页
Journal of Shanghai Jiao tong University:Medical Science
基金
南通市科技局社会发展科技计划(S2008022)~~
关键词
β肌动蛋白
卫星细胞
体外
骨骼肌
快肌
慢肌
小鼠
β-actin
satellite cell
in vitro
skeletal muscle
gastrocnemius
soleus muscle
mouse
