上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (1): 26-.doi: 10.3969/j.issn.1674-8115.2011.01.006

• 论著（基础研究） • 上一篇    下一篇

小鼠快、慢肌卫星细胞的分离培养及β肌动蛋白的表达

朱道立, 陈佩林, 王康乐, 葛 娟, 别 林   

  1. 南通大学 生命科学学院, 南通 226007
  • 出版日期:2011-01-28 发布日期:2011-02-01
  • 作者简介:朱道立 (1951—), 男, 教授；电子信箱: zhudaoli@ntu.edu.cn。
  • 基金资助:

    南通市科技局社会发展科技计划（S2008022）

Culture of mouse satellite cells of gastrocnemius and soleus muscles and expression of β-actin

ZHU Dao-li, CHEN Pei-lin, WANG Kang-le, GE Juan, BIE lin   

  1. School of Life Science, Nantong University, Nantong 226007, China
  • Online:2011-01-28 Published:2011-02-01
  • Supported by:

    Nantong Science and Technology Bureau Foundation, S2008022

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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。

关键词: β肌动蛋白, 卫星细胞, 体外, 骨骼肌, 快肌, 慢肌, 小鼠

Abstract:

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 performed to detect the relative expression of β-actin gene with GAPDH as an internal standard. Results The expression of fast- and slow-myosin was positive in satellite cells of gastrocnemius and soleus cultured in vitro, with favorable cell growth and proliferation. The relative expression of β-actin gene in gastrocnemius and soleus muscles was 3.71072 and 2.106028, respectively (P<0.01). Conclusion The expression of β-actin in gastrocnemius is higher than that in soleus muscle, which suggests the expression of β-actin may be related to the types of muscle fibers.

Key words: β-actin, satellite cell, in vitro, skeletal muscle, gastrocnemius, soleus muscle, mouse