耐钙心肌细胞的分离及其L型钙通道电流观察

Preparation of Rat Cardiomyocytes for Experiment of Patch Clamp and Recording of the Currents of L-type Calcium Channels

目的:建立大鼠心肌细胞分离方法,用于膜片钳实验,并记录L型钙离子通道电流。方法:基于Langendorff逆行主动脉灌流模型,以经改进的心肌细胞分离技术获得单个耐钙心肌细胞,用膜片钳全细胞方式记录L型钙电流。结果:在灌流所得50%存活单个心肌细胞中约70％为耐钙心肌细胞,并记录到典型的L型钙电流。结论:这种心肌细胞分离技术能获得大量具有正常电生理特性的耐钙心肌细胞,可用于心肌细胞离子通道和信号转导机制的研究。

Objective: To devise a method to isolate calcium-tolerant cardiomyocytes from SD rats for the experiment of patch clamp, and record the currents of L-type calcium channels. Methods: Single calcium-tolerant cardiomyocytes were prepared by a modified technique based on Langendorff model of aorta retrograde perfusion apparatus. The currents of L-type calcium channels were recorded by patch clamp in the entire cell mode. Results: There were about 70% single calcium-tolerant cardiomyocytes in all the surviving mono-celled cardiomyocytes. A typical current of L-type calcium channel was recorded. Conclusion: A large number of single calcium-tolerant cardiomyocytes with normal electrophysiological action can be obtained through the isolation technique of cardiomyocytes, which can be used to investigate the mechanism of ion channels and information transmission of cardiomyocytes.