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新生大鼠心肌细胞的分离、培养及微小RNA对其增殖影响的测定

Neonatal Rat Cardiomyocytes Isolation, Culture, and Determination of MicroRNAs' Effects in Proliferation.

作者信息

Tao Lichan, Bei Yihua, Li Yongqin, Xiao Junjie

机构信息

Department of Cardiology, The Third Affiliated Hospital of Soochow University, Changzhou, China.

Cardiac Regeneration and Ageing Lab, School of Life Science, Shanghai University, Shanghai, China.

出版信息

Methods Mol Biol. 2018;1733:203-213. doi: 10.1007/978-1-4939-7601-0_17.

Abstract

Cardiomyocytes loss is a major contributor for many cardiovascular diseases including heart failure and myocardial infarction. Although extremely limited, adult cardiomyocytes are able to proliferate. Understanding the molecular mechanisms controlling cardiomyocytes proliferation is extremely important for enhancing cardiomyocyte proliferation to promote cardiac regeneration and repair. MicroRNAs (miRNAs, miRs) are powerful controllers of many essential biological processes including cell proliferation. Here, we described in detail a protocol for isolation and culture of neonatal rat cardiomyocytes and the determination of miRNAs' effects in proliferation based on two well-established methods including EdU and Ki67 immunofluorescent stainings.

摘要

心肌细胞丢失是包括心力衰竭和心肌梗死在内的许多心血管疾病的主要原因。尽管成年心肌细胞增殖能力极其有限,但仍能够增殖。了解控制心肌细胞增殖的分子机制对于增强心肌细胞增殖以促进心脏再生和修复极为重要。微小RNA(miRNA,miR)是包括细胞增殖在内的许多重要生物学过程的有力调控因子。在此,我们详细描述了一种分离和培养新生大鼠心肌细胞的方法,以及基于EdU和Ki67免疫荧光染色这两种成熟方法来测定miRNA对增殖的影响的方法。

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