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|本期目录/Table of Contents|

成年家兔心房肌细胞的分离和电生理记录(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2008年第5期
页码:
533-536
栏目:
基础研究
出版日期:
2008-10-20

文章信息/Info

Title:
Isolation and electrophysiological recording of adult rabbit atrial myocytes
作者:
马瑞彦1肖颖彬1陈劲进1陈中山2陈鹏慧3程伟1
第三军医大学: 1.新桥医院全军心血管外科中心; 2.西南医院全军眼科中心; 3.基础部神经生物学教研室,重庆 400037
Author(s):
MA Ruiyan XIAO Yingbin CHEN Jinjin CHEN Zhongshan CHEN Penghui CHENG Wei
Department of Cardiovascular Surgery, Xinqiao Hospital, Third Military Medical University Chongqing 400037, China
关键词:
心房肌细胞分离电生理学
Keywords:
rabbit atrial myocytes isolation electrophysiology
分类号:
R329.25
DOI:
-
文献标识码:
A
摘要:
目的 建立一种简单稳定的成年家兔心房肌细胞的分离、培养方法并进行电生理记录。方法 麻醉后取出成年家兔心脏,采用Langendorff灌流装置及急性酶裂解法分离心房肌细胞,差速贴壁法进行纯化后培养于DMEM培养基。在倒置显微镜下观察细胞形态,利用透射电镜观察细胞超微结构,用免疫荧光染色法对心房肌细胞进行鉴定,利用全细胞膜片钳技术记录动作电位和内向钙电流和外向钾电流。结果 本方法分离的心房肌细胞纯度和细胞存活率较高,并使用膜电钳技术成功记录了L型钙电流和瞬时外向钾电流。结论 该方法简便有效,细胞存活率高且为进一步进行各种电生理实验打下了基础。
Abstract:
AIM To construct a simple and stable method of isolating single atrial myoctes in adult rabbits for culturing and various electrophysiological recording. METHODS Single atrial myoctes of adult rabbits were isolated enzymatically by modified Langendorff perfusion technique and were cultured in DMEM culture media after purification. Morphological and ultrastracture were examined by inverted microscope and transmission electron microscope and immunofluorescenct examination was used to identificate the atrial myocytes. Patch clamp was used to record the membrane currents, including ICaL, and Ito current with wholecell recording mode. RESULTS The survival rate and purity were satisfactory and Ltype calcium current and transient outward potassium current were recoded in single atrial myocytes. CONCLUSION This method is simple and effective to get single adult atrial myocytes, which is can be used for further research on electrophysiological properties.

参考文献/References

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[2] Schackow TE, Decker RS, Ten Eick RE. Electrophysiology of adult cat cardiac ventricular myocytes: changes during primary culture[J]. Am J Physiol, 1995, 268(4 pt 1):C1002-C1017.

[3] Mitcheson JS, Hancox JC, Levi AJ, et al. Cultured adult cardiac myocytes: future applications, culture methods, morphological and electrophysiological properties [J]. Cardiovasc Res, 1998, 39(2):280-300.

[4] Zhang R, Yu ZB, Wang YY. \[Isolation of cardiomyocytes from the adult mouse heart\][J]. Sheng Li Xue Bao, 2004, 56(5):656-660.

[5] Piper HM, Volz A, Schwartz P. Adult ventricular rat heart muscle cells[M]// Piper HM. Cell culture techniques in heart and vessel research. Berlin: SpringerVerlag, 1990:36-60.

[6] Bird SD,Doevendans PA, VanRooijen MA, et al. The human adult cardiomyocyte phenotype[J]. Cardiovasc Res, 2003, 58(2):423-434.

[7] Folliguet TA, RucherMartin C, Pavoine C, et al. Adult cardiac myocytes survive and remain excitable during longterm culture on synthetic supports[J]. J Thorac Cardiovasc Surg, 2001, 121(3):510-519.

[8] Messina E, DeAngelis L, Frati G, et al. isolation and expansion of adult cardiac stem cells from human and murine heart[J]. Circ Res, 2004, 95(9):911-921.

备注/Memo

备注/Memo:
收稿日期:2007-01-06.基金项目:国家自然科学基金(30370583) 通讯作者:肖颖彬,教授,主要从事心脏房颤的发病机制及临床治疗研究Email:xiaoyb1@vip.sina.com 作者简介:马瑞彦,讲师,博士Email:maruiyan001@tom.com
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