Li Miao-Ling, Zeng Xiao-Rong, Yang Yan, Liu Zhi-Fei, Ding Yin-Yuan, Zhou Wen, Pei Jie
Department of Myocardial Electrophysiology, Luzhou Medical College, Luzhou 646000, China.
Sheng Li Xue Bao. 2007 Dec 25;59(6):858-64.
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(to1)), 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 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.
为探讨耐钙人心房肌细胞的分离方法,采用蛋白酶(XXIV型)和胶原酶(V型)酶解改良法分离单个心肌细胞。运用全细胞膜片钳技术记录分离的心房肌细胞中的L型钙通道电流(I(Ca-L))、钠电流(I(Na))、瞬时外向钾电流(I(to1))和内向整流钾电流(I(K1))。用该方法分离的单个心肌细胞表面光滑、横纹清晰且呈杆状。可用于电生理研究的存活且耐钙的可记录心肌细胞产量占分离细胞总数的50%-60%。与其他分离方法相比,该方法简单稳定,能获得大量合格的心肌细胞。在这些细胞中记录到的电流具有功能性和活性。我们的研究表明,本文所述方法分离的心肌细胞具有正常的电生理功能,适合进行膜片钳实验。