一种有效急性酶分离大鼠心肌细胞及复钙的方法  

An effective method of acute isolation of cardiomyocytes and calcium recovery in rats

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

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

出　　处：《泸州医学院学报》2016年第1期31-33,共3页Journal of Luzhou Medical College

基　　金：国家自然科学基金资助项目(No:81173661);泸州市-泸州医学院联合专项(2013LZLY-J49)

摘　　要：目的:本研究旨在探讨一种既简便又高效的急性酶分离大鼠心肌细胞及有效复钙的方法,拟建立一种稳定高效分离大鼠心肌细胞且复钙后细胞成活率高的方法,为进一步研究单个心肌细胞的各种生理机制等诸多实验提供有利的前提保障。方法:应用II型胶原酶于Langendorff灌流装置下进行恒压或恒流灌流,得到横纹清晰、立体感较强的大鼠心肌细胞,然后应用三步法对心肌细胞进行复钙。结果:应用Langendorff灌流系统对大鼠心脏进行恒压或恒流灌流II型胶原酶,成功得到横纹清晰、立体感较强的大鼠心肌细胞,且复钙后在台式液中存活率仍然高达50%。结论:成功建立了一种既简便高效又稳定的急性分离大鼠心肌细胞且复钙后心肌细胞成活率较高的方法,为今后研究大鼠心肌细胞舒缩功能及其基本生理特性提供良好的单个细胞。Objective： To establish a simple and efficient method for acute isolation of myocardial cells and effective recovery of calcium in rats, so as to provide a favorable prerequisite protection of cardiomyocytes for further downstream studies of the physiological mechanism of single myocardial cell and many other experiments.Methods： Using type II collagenase in Langendorff irrigation flow device under constant pressure or constant flow perfusion, cardiomyocytes were isolated from rats, followed by a three-step calcium recovery was applied.Results： Cardiomyocytes were successfully isolated from rats using Langendorff irrigation flow system at a constant pressure or constant flow perfusion of collagenase type II, which showed a clear cross striations and strong three-dimensional sense of myocardial cells. After calcium recovery, the survival rate of cardiomyocytes remains as high as 50% in Tyrode solution. Conclusion： A new and high efficient method of acute isolation of cardiomyocytes in rats was established successfully, which can provide functional single cardiomyocyte for further downstream studies.

关 键 词：大鼠 心肌细胞 LANGENDORFF 

分 类 号：R33-33[医药卫生—人体生理学]