大鼠成体心肌细胞的分离和培养  

Isolation and culture of adult rat cardiomyocytes

作　　者：王亭忠[1] 席雨涛[1] 吴格如[1] 马爱群[1] 

机构地区：[1]西安交通大学第一医院心血管内科,教育部环境与疾病相关基因重点实验室,陕西西安710061

出　　处：《第四军医大学学报》2005年第17期1544-1546,共3页Journal of the Fourth Military Medical University

摘　　要：目的: 探索稳定的大鼠成体心肌细胞的分离和培养方法. 方法: 应用主动脉逆行生物酶(5 g/L胰蛋白酶及1 g/L胶原酶)灌流法分离大鼠成体心肌细胞,免疫荧光染色法鉴定分离的心肌细胞,比较有无血清培养对心肌细胞的影响. 结果: 心肌细胞的存活率为95%以上;存活细胞呈杆状,形态完整,长宽比约为4～6∶ 1;无血清培养心肌细胞只能存活1 wk,形态不发生改变;而加血清培养心肌细胞可存活2 wk以上,但超微结构发生了改变,且不同血清浓度对成纤维细胞生长具有不同的作用. 结论: 主动脉逆行生物酶(胶原酶和蛋白酶)灌流法是比较理想的心肌细胞分离方法;血清对成体心肌细胞的培养起着重要作用.AIM： To establish a method of isolation and culture of adult rat cardiomyocytes. METHODS： Adult rat cardiomyocytes were isolated by aorta retrograde collagenase perfusion and identified with immunofluorence of α-actin, desmin and cTnT. The effects of culture with or without serum on adult rat cardiomyocytes were compared. RESULTS： Homogeneous populations of 95% rod-shaped cells were obtained routinely and the cells were complete in shape and the ratio between the length and width was 4 or 6 to 1. Adult rat cardiomyocytes were alive in serum-free culture for only less than 1 weeks and no shape changes were observed, Myocytes in culture with serum were alive for more than 2 weeks but some ultrastructural changes were observed, Serum of different concentrations were found to have different effects on cultured cardiomyocytes. CONCLUSION： Aorta retrograde collagenase perfusion is an ideal method to isolate cardiomyocytes, Serum has some important effect on adult rat cardiomyocytes culture.

关 键 词：大鼠 心肌细胞 分离 培养 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]