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成年小鼠心肌细胞分离技术

张然, 余志斌, 王云英

第四军医大学航空航天生理学教研室,西安,710032

摘要

为进行成年小鼠心肌细胞培养与收缩功能研究,首先必须获得高产量与高质量的心肌细胞.本实验采用Langendorff装置行恒流灌流心脏,同时监测灌流压力的变化.根据小鼠鼠龄微调灌流流速,使初始灌流压力保持在40 mmHg.用0.05%单一粗制胶原酶在37℃条件下消化心脏,当灌流压力下降至28 mmHg时,即刻终止消化。

关键词： 小鼠; 心肌细胞; 分离

Isolation of cardiomyocytes from the adult mouse heart

Zhang Ran, Yu Zhibin, Wang Yunying

Department of Aerospace Physiology, Fourth Military Medical University. Xi'an 710032, China

Abstract

In order to culture cardiomyocytes or to observe the contractile function of adult mouse cardiomyocytes, it is necessary to isolate high-yield and high-quality cardiomyocytes at first. The mouse was injected with heparin (5 000 IU/kg, i.p.) 20 min prior to the experimental protocol, then was sacrificed by cervical dislocation. The heart was excised and the aorta was cannulated rapidly. The cannulated heart was mounted on a Langendorff perfusion apparatus with constant flow and perfusion pressure was monitored. The initial perfusion pressure was maintained at 40 mmHg by regulating the flow rate. The heart was digested by 0.05 % crude collagenase I at 37 oC and the enzymatic digestion was terminated immediately when the perfusion pressure was lowered to 28 mmHg.

Key words: Mice;Cardiac myocytes;Isolation

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引用本文：

张然, 余志斌, 王云英. 成年小鼠心肌细胞分离技术[J]. 生理学报 2004; 56 (5): .

Zhang Ran, Yu Zhibin, Wang Yunying. Isolation of cardiomyocytes from the adult mouse heart. Acta Physiol Sin 2004; 56 (5): (in Chinese with English abstract).