大鼠成肌细胞体外培养及其自发收缩特性  

In vitro culture and auto-contraction of Sprague-Dawley rats’ skeletal myoblasts

作　　者：赵科研[1] 俞世强[1] 蔡振杰[1] 吴铁军[1] 胡军[1] 周和平[1] 

机构地区：[1]第四军医大学西京医院心脏外科,陕西西安710033

出　　处：《第四军医大学学报》2004年第1期34-36,共3页Journal of the Fourth Military Medical University

摘　　要：目的 :体外培养大鼠成肌细胞 ,并详细描述成肌细胞的体外培养的自主收缩特性 .方法 :采用 1 g/L胶原酶消化法分离细胞及非连续密度梯度离心法 (Percoll分离液 )分离纯化细胞 ,在收缩的细胞中加入异丙肾上腺素 (isoproterenol,IP)使终浓度为 0 .1 μmol/L .结果 :成肌细胞融合后 2～ 3d可发生自发收缩 ,大部分频率约 5 0～ 1 6 0 /min ,最快的可达2 0 0 /min .加入IP后细胞收缩频率平均增加约 1 8.4 /min .结论 :成肌细胞具有自发收缩特性 ,且IP对其存在正性变时作用 。AIM: To establish a practical procedure for culturing Sprague Dawley rats’ myoblasts in vitro and to describe the characteristics of these myoblasts. METHODS: Muscle samples were minced and digested by collagenaseⅠand purified with Percoll gradient liquid. Myoblasts were tested by α actin and desmin immunochemistry stains. Many of the myoblasts could auto contract, so isoproterenol (IP) was added into the culture bottles with the final density as 0.1μmol/L so as to compare the contracting rates. RESULTS: The myoblast viability was measured as up to 90 % by trypan blue. Cell proliferation reached the peak 3-5 d after the cells were planted, and cell purity could be as high as 95 % by α actin and desmin immunochemistry stains tests. Myoblastes could contract on the 2nd or 3rd d after they were fused and the contracting rates ranged from 50 to 160 beats per minute for the most part, while some contracted with 200 beats per minute. When isoproterenol was added, the mean contracting rate increased about 18.4 beats per minute compared with that before the adding isoproterenol. CONCLUSION: Myoblasts can auto contract and IP can increase their contracting rates. The transplant of myoblasts into the ischemia heart has been proved to be effective in the treatment of heart diseases.

关 键 词：成肌细胞 细胞培养 收缩 免疫组织化学 

分 类 号：R542.2[医药卫生—心血管疾病]