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一种改进的新生小鼠心肌细胞原代培养方法。

An improved protocol for primary culture of cardiomyocyte from neonatal mice.

作者信息

Sreejit P, Kumar Suresh, Verma Rama S

机构信息

Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600036 TN, India.

出版信息

In Vitro Cell Dev Biol Anim. 2008 Mar-Apr;44(3-4):45-50. doi: 10.1007/s11626-007-9079-4. Epub 2008 Feb 23.

Abstract

The primary culture of neonatal mice cardiomyocyte model enables researchers to study and understand the morphological, biochemical, and electrophysiological characteristics of the heart, besides being a valuable tool for pharmacological and toxicological studies. Because cardiomyocytes do not proliferate after birth, primary myocardial culture is recalcitrant. The present study describes an improved method for rapid isolation of cardiomyocytes from neonatal mice, as well as the maintenance and propagation of such cultures for the long term. Immunocytochemical and gene expression data also confirmed the presence of several cardiac markers in the beating cells during the long-term culture condition used in this protocol. The whole culture process can be effectively shortened by reducing the enzyme digestion period and the cardiomyocyte enrichment step.

摘要

新生小鼠心肌细胞模型的原代培养,除了是药理和毒理学研究的宝贵工具外,还使研究人员能够研究和了解心脏的形态、生化和电生理特征。由于心肌细胞出生后不会增殖,原代心肌培养很困难。本研究描述了一种从新生小鼠快速分离心肌细胞的改进方法,以及此类培养物的长期维持和传代。免疫细胞化学和基因表达数据也证实,在本方案使用的长期培养条件下,跳动细胞中存在几种心脏标志物。通过缩短酶消化时间和心肌细胞富集步骤,可以有效缩短整个培养过程。

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