一种改良的原代大鼠乳鼠心肌细胞分离及培养方法  

An improved method of isolation and culture myocardial cells in neonatal rat

作　　者：程俊[1] 曾晓荣[1] 谭晓秋[1] 李鹏云[1] 文静[1] 陈琳琳[1] 杨艳[1] 

机构地区：[1]西南医科大学心血管医学研究所医学电生理教育部重点实验室,四川泸州646000

出　　处：《四川生理科学杂志》2016年第4期187-189,共3页Sichuan Journal of Physiological Sciences

基　　金：国家自然科学基金资助项目(编号:81173661);泸州市-泸州医学院联合专项(编号:2013LZLY-J49)

摘　　要：目的:本研究旨在探讨一种经改良后既稳定又高效的分离大鼠乳鼠心肌细胞的原代分离和培养方法,拟建立此种方法,为进一步研究单个心肌细胞上的各种电生理机制等诸多实验提供有利的前提保障。方法:取出生1-3天内SD大鼠心脏,联合应用胰蛋白酶和Ⅱ胶原酶两步消化,离心收集到心肌细胞,差速培养,加入5-溴脱氧尿嘧啶核苷纯化后得到原代心肌细胞,经高糖DMEM培养,即可得到立体感较强的大鼠乳鼠心肌细胞。结果:联合应用胰蛋白酶和Ⅱ胶原酶两步消化,经过高糖DMEM培养后得到的心肌细胞存活率高、纯度高、折光性好、立体感较强,有节律性搏动。结论:成功建立了一种既简便高效又稳定的分离大鼠乳鼠心肌细胞的方法,为今后研究乳鼠心肌细胞的功能及其基本电生理特性提供良好的单个细胞,可广泛应用于各种心血管疾病的研究中。Objective：To investigate a stable and efficient isolation and culture rat myocardial cells in rat neonatal,to provide premise and guarantee for many experiments to study various kinds of electrophysiological mechanism on single myocardial cells.Methods：Rat myocardial cells were digested using two step method with trypsin and collagenase II in newborn SD rat aged 1-3days.All collected myocardial cells were differential cultured,added 5-5-bromodeoxyuridine,and cultured in high glucose DMEM to get the strong three-dimensional sense of rat myocardial cells.Results：Myocardial cells with high survival rate,high purity,good refraction,good refraction,strong dimensional sense and rhythmic beat could be obtained after trypsin and collagenase digestion and high glucose DMEM cultivation.Conclusion：We have successfully established a simple effective and stable method for the isolation of neonatal rat cardiac muscle cells,which provides a good single cell for the future study of the function and basic electrophysiological properties of neonatal rat cardiac muscle cells,which can be widely used in various heart disease studies.

关 键 词：乳鼠 心肌细胞 

分 类 号：R54[医药卫生—心血管疾病]