新生大鼠心肌细胞的分离与培养  

Isolation and culture of cardiomyocytes from neonatal rats

作　　者：王现涛[1] 李浪[1] 苏强[1] 孙羽涵[1] 杨华锋[1] 

机构地区：[1]广西医科大学第一附属医院心内科广西心血管病研究所,南宁530021

出　　处：《中华老年心脑血管病杂志》2015年第7期745-747,共3页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：广西优秀博士学位论文培育项目(YCBE2014023)

摘　　要：目的探讨新生大鼠心肌细胞分离与培养的条件和方法,建立简单、可靠的原代心肌细胞培养方法。方法新生SD乳鼠10只,开胸取心剪碎后,胰蛋白酶和Ⅱ型胶原酶联合消化心肌组织,差速贴壁法纯化心肌细胞。倒置荧光相差显微镜下连续观察15 d心肌细胞形态特征及变化。用0.4%锥虫蓝染色测心肌细胞存活率,记录心肌细胞搏动次数。结果心肌细胞24 h基本贴壁并伸出伪足,出现自发性搏动,72 h后心肌细胞逐渐形成细胞簇,并呈现部分同步搏动,之后心肌细胞连接成片生长。心肌细胞存活率为96.5%,培养72 h至第15天内心肌细胞搏动次数无差异(P>0.05)。结论用该法分离与培养新生大鼠心肌细胞,操作简单且稳定可靠,心肌细胞存活率较高。Objective To establish a simple and reliable technique to culture primary cardiomycytes from neonatal rats by studying their isolation and culture.Methods Cardiac tissue taken from 10 neonatal rats was digested with trypsinase plus type Ⅱ collagenase.The cardiomycytes were purified by differential adherent culture and their morphology was observed under inverted fluorescence microscope for 15 days.The survival rate of cardiomycytes was assayed with 0.4%Trypan blue staining.The beating times of cardiomycytes were recorded.Results The cardiomycytes were adhered with spontaneous pulsation and their parapodia were stretched out after 24 hours of culture followed by synchronous pulsation after 72 hours of culture,and then grew in a patchy manner.The survival rate of cardiomycytes was 96.5%.No significant difference was found in the beating times.Conclusion The technique we established is simple and stable for isolation and culture of cardiomyocytes from neonatal rats.The survival rate of cardiomyocytes is rather high.

关 键 词：肌细胞 心脏 胰蛋白酶 胶原酶类 存活率 细胞 培养的 模型 动物 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]