小鼠胚胎心肌细胞分离及电生理特性.pdfVIP

生理学报 Acta Physiologica Sinica, October 25, 2004, 56 (5): 651-655 651 骆红艳等: 小鼠胚胎心肌细胞的分离及电生理特性 实验技术 小鼠胚胎心肌细胞的分离及电生理特性 * 骆红艳, 唐 明, 胡新武, 宋明柯, 梁华敏, 杜以梅, 张 毅 华中科技大学同济医学院生理系, 武汉 430030 摘 要: 本文旨在探索小鼠胚胎心肌细胞的分离方法并观察其电生理特性。应用胶原酶B 消化法获得不同时期单个小鼠胚胎 心肌细胞; 利用全细胞膜片钳技术, 记录胚胎心肌细胞的超极化激活的非选择性内向阳离子电流(If)和 L- 钙电流( ICa-L ), 并用电流 钳记录其自发性动作电位。胚胎心肌细胞通过相差显微镜依据其形态和自发性收缩进行鉴定。本法分离所获得的胚胎心肌 细胞容易进行全细胞膜片钳记录, 可用于记录I f 、I Ca-L 电流和自发性动作电位, 已证实胚胎心肌细胞I f 和I Ca-L 的电生理特性与 成年起搏细胞或心肌细胞相似。本实验建立的分离方法简单、稳定、有效、可靠, 最早可获得 8.5 d 的胚胎心肌细胞。胚 胎心肌细胞的电生理记录为探索胚胎心肌细胞的电生理特性提供了一个可用的模型, 并可能为某些心脏疾病产生的机制提供实 验依据。 关键词: 胚胎; 心肌细胞; 小鼠; 分离 中图分类号: Q463; R329 Isolation and electrophysiological characteristics of embryonic cardiomyocytes in mice * LUO Hong-Yan, TANG Ming , HU Xin-Wu, SONG Ming-Ke, LIANG Hua-Min, DU Yi-Mei, ZHANG Yi Department of Physiology, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China Abstract : To explore the electrophysiological characteristics of embryonic cardiomyocytes, single embryonic cardiomyocytes were obtained from mice at different periods by a collegenase B digestion approach, whole cell patch clamp recording technique was used to record If and ICa-L, and spontaneous action potential was also recorded. The morphological and spontaneous contractile properties of the isolated cells appeared to be typical embryonic cardiomyocytes when the cells were assessed by phase-contrast microscope. Whole cell recording of isolated cells is easily performed by the whole cell patch clamp technique. Elelctrophysiolgical properties of If and Ica-L from embryonic cardiomyocytes have been proved to be similar to those from adult pacemaker cells or cardiomyocytes. The established method of isolation is simple, stable, effective and reliable. It allows to obtain as ear