BALB/c小鼠乳鼠心肌细胞的原代培养

The primary culture of cardiomyocytes from neonatal BALB/c mice

目的:建立分离、纯化和培养BALB/c小鼠乳鼠心肌细胞的方法。方法:取出生1～3天以内的BALB/c小鼠乳鼠心肌组织,用0.125%胰蛋白酶和0.1%Ⅰ型胶原酶消化,将心肌细胞收集到含15%胎牛血清的DMEM培养基中,用差速贴壁分离法和5-溴脱氧尿苷相结合分离、纯化心肌细胞。采用α-actin抗体进行免疫组化染色鉴定心肌细胞,用台盼蓝染色检查心肌细胞成活率。结果:24小时后可见部分单个心肌细胞搏动,48小时后可见心肌细胞相互连接同步搏动;免疫组化染色后可见培养的细胞是心肌细胞,心肌细胞成活率达95%。结论:本研究应用的方法可获得状态好、高成活率的BALB/c小鼠心肌细胞,是一种可靠的心肌细胞培养方法。

Objective：To establish a method for isolating,purifying and culture cardiomyocytes from neonatal BALB/c mice.Methods：Myocardial tissues from neonatal BALB/c mice born within 1 to 3 day were digested with 0.125% trypsin and 0.1% collagenaseⅠ.The cardiomyocytes were collected in DMEM medium,which contains 15% fetal bovine serum.Purification of cardiomyocytes was done by differential adhesion and the supplement of 5-bromodeoxyuridine.Cardiomyocytes were identified by immunohistochemistry staining of α-actin and we stained Cardiomyocytes with trypan blue.Results：After 24 hours some single cardiomyocytes began to beat spontaneously.After 48 hours,cardiomyocytes got together and beat synchronously.Cells used in experiments were confirmed as cardiomyocyte and 95 percent were alive.Conclusion：High quantity and survival cardiomyocytes of BALB/c mice can be effectively and reliably cultured by this method.