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目的建立单个人心房肌细胞的急性分离技术,以满足检测人心肌细胞离子通道的需要。方法两步酶解法消化分离人心房肌细胞,采用膜片钳全细胞技术记录瞬时外向钾电流(Ito)。结果根据患者的性别、年龄及组织块的大小,选择适当的消化时间,分离得到结构清晰,横纹清楚,边缘整齐,少颗粒,舒展伸直,呈长杆状的单个心肌细胞;采用全细胞膜片钳技术能记录到瞬时外向钾电流。结论该方法能分离到形态和状态均良好的细胞,有利于进行单个人心房肌细胞膜片钳实验的研究。
Objective To establish an acute isolation technique of single atrial myocyte in order to meet the need of detecting ion channels of human cardiomyocytes. Methods Two-step enzymatic digestion of human atrial myocytes was performed and transient outward potassium current (Ito) was recorded using patch-clamp whole cell technique. Results According to the patient’s sex, age and the size of the tissue block, the appropriate digestion time was selected to obtain a single cardiomyocyte with clear structure, clear stripes, neat edges, few grains, stretched straight, Patch-clamp technology records transient outward potassium current. Conclusion This method can separate the cells with good morphology and state, and is beneficial to the patch clamp experiments of single human atrial myocytes.