A simple technique for isolating healthy heart cells from mouse models

被引:117
作者
Shioya, Takao [1 ]
机构
[1] Saga Univ, Fac Med, Dept Physiol, Saga 8498501, Japan
关键词
mouse; cardiac myocyte; contraction; action potential; ionic current;
D O I
10.2170/physiolsci.RP010107
中图分类号
Q4 [生理学];
学科分类号
071003 [生理学];
摘要
Single heart cells of mouse models provide powerful tools for heart research. However, their isolation is not easy, and it imposes a significant bottleneck on their use in cellular studies of the heart. Aiming to overcome this problem, this report introduces a novel technique that reproducibly isolates healthy heart cells from mouse models. Using simple devices that ensure easy handling and the rapid aortic cannulation of a small mouse heart, cell isolation was done under physiological conditions without using the "KB" medium or 2,3-butanedione monoxime (BDM). The isolated cells consistently had a healthy appearance and a high viability of 75 +/- 5% (mean +/- SD) in Tyrode solution containing 1.8 mm Ca2+. After 8 h of storage at 37 degrees C, they still had a viability of 45 +/- 12%. The cells showed normal contraction properties when field-stimulated, and they generated normal action potentials and membrane currents under the whole-cell clamp condition. The beta-adrenergic signal transduction of the cells was also normal when it was examined with the isoproterenol enhancement of the L-type Ca2+ current.
引用
收藏
页码:327 / 335
页数:9
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