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目的:研究用于膜片钳实验的SD大鼠心室肌细胞的分离技术,并记录心室肌细胞L型钙通道的电流。方法:采用改进的Langendorff酶液灌流技术分离获得单个心室肌细胞;采用全细胞膜片钳技术记录L型钙通道电流。结果:分离所得70%～80%的存活心肌细胞中有60%～70%左右的耐钙心肌细胞;并记录到典型的L型钙通道电流,其激活电位为-30mv,最大激活电压为0mv。结论:该方法能够分离出足量的可以进行膜片钳实验的心室肌细胞。
OBJECTIVE: To study the isolation technique of SD rat ventricular myocytes used in patch-clamp experiments and record the current of L-type calcium channels in ventricular myocytes. Methods: Single ventricular myocytes were isolated by modified Langendorff enzymatic perfusion technique. L-type calcium channel current was recorded by whole-cell patch-clamp technique. RESULTS: About 60% -70% of calcium-resistant cardiomyocytes were isolated from 70% to 80% of surviving cardiomyocytes. A typical L-type calcium channel current was recorded with an activation potential of -30mv and a maximum activation voltage of 0mv . Conclusion: This method can isolate enough ventricular myocytes to perform patch-clamp experiments.