改良的乳鼠原代心房肌细胞的培养及鉴定方法

目的:主要探讨原代心房肌细胞的培养及鉴定方法,为进一步研究心房颤动的重构机制及治疗方法奠定基础。方法:选取1-3 d的SD乳鼠40只,雌雄不限,分离心房、心室肌,胰酶联合EDTA充分消化心房肌细胞,利用心房肌与成纤维细胞的差速贴壁及细胞传代方法纯化心房肌细胞,免疫细胞化学染色鉴定心房肌细胞。结果:心房肌细胞培养至第3天,可见心房肌细胞覆盖率高达90%,并出现波动性,免疫细胞化学染色可见90%的心房肌细胞肌经α-肌动蛋白抗体染色阳性。结论:经酶化学消化法可成功培养出原代心房肌细胞,是一种较好的培养及鉴定乳鼠心房肌细胞的方法。

Improved Primary Culture and Identification of Atrial Myocytes in Neonatal Rat

Objective:To investigate the culture and identification of major primary atrial myocytes, which can be used to study the remodelling of atrium or relative future research.Methods:Atrial and ventricular muscle is isolated from 40 Sprague Dawley rats(1-3 days old, male or female), trypsin EDTA is added to digest fully, then atrial myocytes were purified with differential adhesion method and passage and identified with immunocytochemistry.Results:Atrial myocytes coverage are up to 90% after 72 hours primary culture, and show the feature of volatility, 90% of myocytes of atrium can be authenticated by alpha-actin antibody staining with immunocytochemistry.Conclusion:Atrial myocytes with high purity and vigor can be separated by enzyme digestion, it is a better method of cultivation and identification of atrial myocytes of neonatal rat.