题名页.docVIP

题名页 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，宋桂利，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 通讯作者：唐明 地址：湖北省武汉市桥口区航空路13# 电话：027传真：+86-27E-mail: tangming49@ 中图分类号：R329 总页数：9 图数：4 文章字数：3000 This work was supported by the National Natural Science Foundation of China (No. 本研究工作受国家自然科学基金资助（No。 Corresponding author. Tel: +86-27 Fax: 86-27 E-mail: tangming49@ 小鼠胚胎心肌细胞的分离及电生理特性 骆红艳唐 明*胡新武宋桂利梁华敏杜以梅张 毅 华中科技大学同济医学院生理系，武汉，430030 摘 要摸索小鼠胚胎心肌细胞分离方法并观察其电生理特性。应用胶原酶B消化法可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），并用电流钳可记录其自发性动作电位。胚胎心肌细胞可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If、ICa-L电流和自发性动作电位，已证实胚胎心肌细胞If和ICa-L的电生理特性与成年起搏细胞或心肌细胞相似。本实验所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有用的模型，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：R329 Isolation and electrophysiological characteristics of embryonic cardiomyocytes in mice LUO Hong-Yan, TANG Ming*, HU Xin-Wu, SONG Gui-Li, 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, the single embryonic cardiomyocyte was obtained from mice at different period with collegenase B digestion method and whole cell patch clamp recording technique was used to record If and ICa-L and spontaneous action potential was recorded. The morphological and spontaneous contractile properties of isolated cell appeared to be typical embryonic cardiomyocytes when assessed by phase-contrast microscope. Whole cell recording of isolated cells was easily performed by the whole cell patch clamp technique. Spontaneous action potentials have been recorded. 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 was simple, stable,