Objective: To explore a simple and practical method for isolating adult mice cardiac myocytes and observe the calcium current characteristic of the myoeytes.Methods: Perfusion from inferior vena cava, aortic cannulation in situ and eollagenase Ⅱ digestive methods were used to obtain single cardiomyocyte.These myocytes were observed by inverted microscope, the myocytes viability was evaluated by trypan bule staining, and the membrane calcium current was recorded by the whole cell patch clamp technique.Results:Rod-shaped, clear-striped, refractive, quiescent, and adherent myocytes with generous high viability were gotten.After step exposed to normal calcium concentration solution, 50％ ～60％ calcium-resistant myocytes were left.Typical L-type calcium current was easily achieved and disappeared after exposed to Tyrode's solution with nitrendipine.Conclusion: It is a simple convenient and reproducible method to isolate adult mice eardiomyocytes with nomal electrophysiological characteristics.%目的:探索建立简单实用的小鼠心肌细胞分离方法并观察其钙电流特征,以便有利于心肌细胞电生理和分子生物学研究.方法:采用下腔静脉灌流,原位主动脉插管和胶原酶Ⅱ消化法得单个心肌细胞.在倒置显微镜下观察细胞形态,台盼蓝染色判定心肌细胞活力,利用全细胞膜片钳技术记录钙电流.结果:分离得到的心肌细胞数量多、存活率高、杆状、横纹清晰、折光性好、静止、贴壁良好.逐步复钙后可获得50%～60%耐钙细胞,单个心肌细胞在全细胞膜片钳模式下可记录到典型的L-型钙电流,在细胞外液加入尼群地平后L-型钙电流消失.结论:该心肌细胞分离方法简单实用,重复性好,所获得的单个心肌细胞具有正常的电生理活性.
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