膜片钳技术研究中钙耐受的心室肌细胞分离方法

Patch-clamp Technique in the Study of Calcium Tolerant Ventricular Myocytes Separation Method

目的：探索并建立适合于膜片技术研究的心室肌细胞急性酶分离方法。方法：采用改进的自制的Langendorff心脏灌流系统,用无钙Tyrode液和酶解液逆行主动脉灌流之后,分次剪取心肌组织置于细胞保存液（KB液）中保存以供全细胞膜片钳方式记录膜电位和膜电流。结果：分离所得80%-90%的细胞呈杆状,边缘清楚,表面光滑,纹理清晰,可记录到典型的动作电位。结论：控制好分离心室肌细胞的实验条件以及熟练掌握实验操作的方法,是分离出适合于膜片钳实验的心室肌细胞的重要因素。

Objective ：To explore and establish a simple and reliable isolation method of single myocardial cell in animols for patch clamp experiments. Methods：Hearts were firstly perfused by the Langendorff perfusion apparatus with Ca^2＋ -free Tyrode＇S solution and enzyme solution retrogradated arteriae aorta. The myocardial tissues were minced into small pieces in KB solution, and conserved in KB solution. The myocardial cells were supplied to record electric current and ventricular myocytes through the way of whole cell patch clamp. Results：The survival animols of myocardial ceils was 80% -90% and the morphous and construction of myocardial ceils was fine. The ventricular myocytes was recorded successfully. Conclusion ： Control the separation of myocardial cell experimental conditions, as well as proficiency in the experimental operation, is an important factor in isolated myocardial ceils suitable for patch clamp experiments.