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2TMR Integrative Medicine  2019, 3: e19001.       DOI: 10.12032/TMRIM201903001
    
A method for isolating rat cardiomyocytes by acute enzymatic hydrolysis
Bo Liang1,2, Hui-Ling Liao1,2,*()
1School of Integrated Traditional Chinese and Western Medicine, Luzhou, China
2Cardiovascular Department, Hospital (TCM) Affiliated to Southwest Medical University, Luzhou, China
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Highlights:

This research was introduced a method of isolate rat cardiomyocytes though acute enzymatic hydrolysis, provides a new direction for clinical research.

Abstract

Objective: This research showed a method of isolate rat cardiomyocytes though acute enzymatic hydrolysis. Method: Our method was divided into five steps: preparation, heart extraction, perfusion, mechanical dissociation and purification and recalcification. Firstly, make a preparation, including solutions, equipment and so on. Secondly, after anesthesia satisfactorily, open the chest of rat and take the heart out. Thirdly, transect the aorta and hang the aorta onto the Langendroff perfusion system such that the tip is just distal to the aortic valve. Forth, perfuse the heart with digestion solution for 17-20 min after Ca2+ free tyrode solution for 5 min. Finally, remove the heart from the system quickly and dissociate mechanically in the KB solution and recalcification after filter. Result: Though the microscope, good-state cardiomyocytes (clear stripes, strong sense of three-dimensional, stick well) can be observed. Conclusion: Those cells can be used to undergo medical electrophysiological experiment, including the study of iron channel.



Key wordsCardiomyocytes      Acute isolation      Acute enzymatic hydrolysis      Patch clamp      Myocardial electrophysiology     
Published: 14 January 2019
Corresponding Authors: Liao Hui-Ling   
E-mail: 15181400995@qq.com
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Effects of hypoxia on sodium current in rat cardiomyocytes [2TMR Integrative Medicine. 2018, 2 (4): 192-196. ]
Cite this article:

Bo Liang, Hui-Ling Liao. A method for isolating rat cardiomyocytes by acute enzymatic hydrolysis. 2TMR Integrative Medicine, 2019, 3: e19001.  

URL:

https://www.tmrjournals.com/im/EN/10.12032/TMRIM201903001     OR     https://www.tmrjournals.com/im/EN/Y2019/V3/IVolume 3/88888

Figure 1 Langendroff perfusion system
Solution Composition pH
Ca2+ free Tyrode solution (mM) NaCl 135
KCl 5.4
MgCl2 1
NaH2PO4 0.33
D-glucose 5
HEPES 10
7.4 with NaOH
Digestion solution Ca2+ free Tyrode solution
collagenase 0.5g / L
bovine serum albumin 1 g / L
7.4 with NaOH
KB solution (mM) KOH 80
KCl 40
KH2PO4 25
MgSO4 3
glutamic acid 50
taurine 20
EGTA 0.5
HEPES 10
glucose 10
7.2 with KOH.
Table 1 The relevant solution of this treatment
Figure 2 The good-state cardiomyocytes with clear stripes and strong sense of three-dimensional can be observed
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[1] Bo Liang, Hui-Ling Liao. Effects of hypoxia on sodium current in rat cardiomyocytes[J]. 2TMR Integrative Medicine, 2018, 2(4): 192-196.