新生大鼠心肌细胞原代培养方法的改良  

The primary culture of neonatal rat cardiac cells

作　　者：李博[1] 吴慧颖[1] 朴虎林[1] 柳克祥[1] 

机构地区：[1]吉林大学第二医院,吉林长春130041

出　　处：《中国实验诊断学》2012年第2期200-203,共4页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技厅科技发展计划项目资助(200905144)

摘　　要：目的探讨酶法分离新生大鼠心肌细胞,提高心肌原代细胞存活率及纯度。方法应用0.1%的Ⅱ型胶原酶消化出生第1d乳鼠的心肌组织,收集的细胞用含10%胎牛血清的DMEM培养基中和,加入DNA酶消化一次,用差速贴壁分离法分离。结果在分离培养新生大鼠心肌细胞过程中,胶原酶和DNA酶的组合使用,并改变差速贴壁时间,可得到高存活率和高纯度的心肌细胞,细胞搏动率高,持续时间久。结论本研究应用改良的新生乳鼠心肌细胞培养方法,心肌细胞存活率高,纯度高,且操作简便,重复性好,是一种较为理想的心肌细胞原代培养方法,可满足实验要求。Objective To investigate how to enzyme isolate the cardiomyocytes of neonatal rat,and improve survival rate of primary cardiac cells and purity of isolated cells.Methods Digest 0.1% collagenase type Ⅱ to born the first 1 d neonatal rat cardiac tissue.With 10% fetal bovine serum DMEM neutralizes the collecting cells.Add a DNA enzyme to digest it,and then separate it by differential adhesion separation.Results During the process of isolating and culturing rat cardiac cells,a combination use of collagenase and DNA enzymes and change the differential adhesion time can get high survival rate and high purity of myocardial cells which has high pulse rate and long duration.Conclusion This study applies the improved cell culture of neonatal rat cardiac cell,so that it has higher survival rate of myocardial cells,high purity,simple operation and good repeatability.It is an ideal method for primary culture of myocardial cells.It can meet the experimental requirements.

关 键 词：心肌细胞 原代培养 胶原酶 分离 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]