A modified method for isolation of rat cardiomyocytes and recording of potassium,sodium and calium current

LONG Yi; ZHOU Ya-nan; YIN Yue-hui

doi:10.13406/j.cnki.cyxb.2011.08.012

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A modified method for isolation of rat cardiomyocytes and recording of potassium,sodium and calium current

更新时间：2024-01-17

- A modified method for isolation of rat cardiomyocytes and recording of potassium,sodium and calium current
- Journal of Chongqing Medical University Vol. 36, Issue 8, Pages: 910-912(2011)
- 作者机构：
  
  重庆医科大学附属第二医院心血管内科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2011.08.012
  CLC： R329
- Published：2011，
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[1]龙毅,周亚南,殷跃辉.急性分离大鼠心房肌细胞方法的改进及对钾、钠、钙电流的记录[J].重庆医科大学学报,2011,36(08):910-912.

LONG Yi, ZHOU Ya-nan, YIN Yue-hui. A modified method for isolation of rat cardiomyocytes and recording of potassium,sodium and calium current. [J]. Journal of Chongqing Medical University 36(8):910-912(2011)
[1]龙毅,周亚南,殷跃辉.急性分离大鼠心房肌细胞方法的改进及对钾、钠、钙电流的记录[J].重庆医科大学学报,2011,36(08):910-912. DOI： 10.13406/j.cnki.cyxb.2011.08.012.

LONG Yi, ZHOU Ya-nan, YIN Yue-hui. A modified method for isolation of rat cardiomyocytes and recording of potassium,sodium and calium current. [J]. Journal of Chongqing Medical University 36(8):910-912(2011) DOI： 10.13406/j.cnki.cyxb.2011.08.012.

摘要

目的:介绍1种改进的成年SD大鼠心房肌细胞分离方法。方法:SD大鼠麻醉后

快速取出心脏

将主动脉结扎在Langendorff灌流装置上

无钙台式液灌流5 min后换用酶液灌流10～12 min

剪取心耳组织于KB液中剪碎并吹打

200目细胞筛网过滤

将细胞滤液500 r离心2 min

梯度复钙后将细胞悬液置于4℃冰箱中静置1 h

应用全细胞膜片钳技术记录大鼠心房肌细胞瞬时外向钾电流（Ito）、快钠电流（INa）以及L型钙电流（ICa-L）。结果:本方法分离出的心房肌细胞存活率高

结构完整

具有良好的耐钙性

可成功记录出Ito、INa、ICa-L。结论:改进的实验方法可获得大量耐钙心肌细胞

并具有正常的电生理特性。

Abstract

Objective:To introduce a modified isolation method for acquiring rat atrial myocytes.Methods:The heart of adult sprague-dawley(SD) rat was rapidly excised after successful anesthesia and mounted on a Langendorff apparatus.It was perfused retrogradely via the aorta for five minutes with Ca2+-free Tyrode’s solution and then perfused with collagenase dissolved in Tyrode’s solution for ten to twelve minutes

and then the atrial appendage was sheared and stored in a kraftbruhe(KB) solution.Thereafter tissue was minced and filtrated.After being centrifuged at 500 r for 2 min

the isolated myocytes were performed gradient calcium reintroduction and allowed to pellet via gravity for 1 hour in refrigerator

then the transient outward potassium current

sodium current and L-tpye calium current were recorded by the use of patch clamp whole cell recording technique.Results:The survival rate of atrial myocytes was high and the morphous and construction of atrial myocytes were fine and the property of calcium-tolerance was good.The transient outward potassium current

sodium current and L-tpye calium current were recorded successfully.Conclusion:The modified isolation method can produce large numbers of single calcium-tolerance myocardial cells and normal electrophysiological properties.

关键词

膜片钳心肌细胞瞬时外向钾电流钠电流L型钙电流细胞分离

Keywords

patch clampatrial myocytestransient outward potassium currentsodium currentL-type calium currentcell isolation

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Department of Gastroenterology, Second College of Clinical Medicine, Chongqing Medical University

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