摘要
目的:探索建立简单可靠的心肌细胞的酶解分离方法,提高细胞存活率和膜片钳实验成功率。方法:采用Langendorff装置,无钙液和胶原酶P二步灌流法分离心肌细胞。显微镜下观察细胞形态结构,并应用膜片钳全细胞技术记录大鼠心室肌细胞膜瞬时外向钾电流。结果:心肌细胞存活率高,形态结构良好,可成功记录出瞬时外向钾电流。结论:通过改进的分离方法获得的心肌细胞存活率高,具有正常的电生理特性。
Objective: To explore and establish a simple and reliable method of isolating single cardiac myocytes in order to improve the quantity and quality of cells for patch clamp research.Methods: The ventricular myocytes were isolated enzymatically by two-step perfusion with calcium-free solution and collagenase P on the Langendorff apparatus.The morphous and construction were observed under the microscope,and the transient outward potassium current was recorded by the use of patch clamp whole-cell recording tech...
基金
国家自然科学基金(No.30672465)
广东省自然科学基金重点项目(No.06118928)
广东省医学科研基金(A2008435)
关键词
心肌
细胞分离
膜片钳
瞬时外向钾电流
myocardium
cell isolation
patch clamp
the transient outward potassium current
