Balb/c乳鼠心肌细胞的分离和体外培养方法的优化  

Optimization for the separation and cultivation of suckling Balb/c mouse myocardial cell in vitro

作　　者：张中海[1] 包琳[1] 史巧云[1] 李晓波[1] 钟照华[1] 

机构地区：[1]哈尔滨医科大学微生物学教研室,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2007年第4期324-326,共3页Journal of Harbin Medical University

基　　金：哈尔滨市科技攻关计划资助项目(2005AA9CS116-16);黑龙江省教育厅科学技术研究资助项目(11511159)

摘　　要：目的探讨Balb/c乳鼠心肌细胞分离和培养方法,优化出Balb/c乳鼠心肌细胞分离和培养的方法。方法取乳鼠心脏,采用胰酶和胶原酶Ⅱ消化方法分离乳鼠心肌细胞,比较消化次数和消化时间对原代乳鼠心肌细胞培养的影响。结果原代培养的Balb/c乳鼠心肌细胞生长迅速、自行搏动,细胞形态完整,细胞内肌丝、线粒体清晰。分离细胞时消化次数最好控制在不多于6次,每次消化时间不超过20min。2h差速贴壁可以提高心肌细胞纯度,经肌动蛋白鉴定,心肌细胞达94.6%。结论本研究优化了分离和原代培养Blab/c乳鼠心肌细胞的方法。Objective To optimize the protocol for separation and cultivation in vitro of suckling Balb/c mouse myocardial cells.Methods The suckling Balb/c mouse heart tissues were obtained and digested with trypsin and coagulase Ⅱ . The effects of digestion procedures on the primary cultivation of suckling Balb/c mouse myocardial cells had been compared. Results The primary cultured myocardial cells grew well and could contract autonomously in vitro. It kept intact cellular shape, filaments, and mitochondria. The digestions should be less than 6 times, and every digestion should last for less than 20 min. Two-hour differential adherence could improve the purity of myocardial cells. By this protocol, 94.6 % of cultured cells were live myocardial cells. Conclusion The protocol for separation and cultivation of suckling Balb/c mouse myocardial cells are optimized.

关 键 词：心肌细胞 Balb/c乳鼠 原代细胞培养 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]