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An improved method for isolation of single atrial myocyte from human heart

Li Miaoling, Zeng Xiaorong, Yang Yan, Liu Zhifei, Ding Yinyuan, Zhou Wen, Pei Jie

Department of Myocardial Electrophysiology, Luzhou Medical College.Luzhou 646000,Sichuan

Abstract

To approach the method of isolation of tolerant human atrial myocytes, single myocytes were isolated by modified procedure of enzymatic dissociation with protease （type XXIV） and collagenase （type V）. L-type calcium channel current （I_(Ca-L)）, sodium current （I_(Na)）, transient outward potassium current （I_(tol)）, and inward rectifier potassium current （I_(K1)） in isolated atrial myocytes were recorded by using whole-cell patch-clamp techniques. Single cardiocytes isolated by this method were smooth, well-striated and rod-shaped. The yields of recordable myocytes, which were viable and calcium-tolerant for electrophysiological studies, were 50%-60% of the total isolated cells. Compared with other isolation methods, this method was simple and steady, but with yield of a great number of qualified myocytes. The currents recorded in these cells were functional and active. Our research suggests that the myocytes isolated by the described method in this paper have normal electrophysiological function and are appropriate for patch-clamp experiments.

Key words: atrial myocyte;cell separation;Patch-clamp techniques;ionic channel

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Citing This Article：

Li Miaoling, Zeng Xiaorong, Yang Yan, Liu Zhifei, Ding Yinyuan, Zhou Wen, Pei Jie. An improved method for isolation of single atrial myocyte from human heart. Acta Physiol Sin 2007; 59 (6): 858-864 (in Chinese with English abstract).