犬左心房心肌细胞的分离方法及电生理记录

Cell isolation method and electrophysiological recording for canine left atrial cardiomyocytes.

目的探索简单、稳定的犬左心房肌细胞的分离方法。方法采用改良的Langendorff灌流法及自制的冠脉插管装置。对分离得到犬左心房肌细胞进行形态学观察和应用膜片钳技术记录动作电位和瞬间外向整流钾电流(Ito)以验证其电生理特性。结果分离状态良好的单个左心房细胞率为70%～80%,呈杆状,边缘锐利,横纹清晰;电生理记录显示具备正常的心房肌细胞电生理特性。结论该方法简单,可节约实验成本和时间,分离的活细胞产量高,质量好,值得推广使用。

Objective To explore an easier and reliable technique for isolation of myocytes from left atrium of dogs. Methods Modified Langendorff perfusion method and home-made coronary catheterization devices were used in the experiments.Morphologic observation and electrophysiological study via recording action potentials and transient outward rectifier K＋ current（Ito） using patch-clamp technique were carried out in isolated canine left atrial cardiomyocytes. Results Specific cardiomyocytes,which presented as rod-shaped,viable and quiescent cells with normal electrophysiological properties,accounted for 70%~80% of the isolated cells using the modified method. Conclusion This method is simple and can save experimental cost and time with high yield of good quality cardiomyocytes.