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An improved protocol for primary culture of cardiomyocyte from neonatal mice

  • P. Sreejit
  • Suresh Kumar
  • Rama S. Verma
Article

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.

Keywords

Primary cell culture Neonatal mice Murine cardiomyocyte enrichment Immunostaining Gene expression 

Notes

Acknowledgments

This work is supported by grants to R.S.V. by the Ministry of Human Resource Development (MHRD—BIO/2005–2006/007/MHRD/RAMS/859) and Department of Biotechnology, Ministry of Science and Technology (DBT-BT/PR5392/MED/14/693/2004).

Supplementary material

Supplementary data 1A

Cardiomyocte beating in cluster (MPG 1.15 MB)

Supplementary data 1B

Individual beating cardiomyocytes (MPG 988 kb)

Supplementary data 1C

Single beating cardiomyocyte (MPG 982 kb)

Supplementary data 1D

(MPG 1.42 MB)

11626_2007_9079_MOESM5_ESM.doc (30 kb)
Supplementary Table 1 PCR primers used for the study (DOC 30.5 KB)

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Copyright information

© The Society for In Vitro Biology 2008

Authors and Affiliations

  1. 1.Stem Cell and Molecular Biology Laboratory, Department of BiotechnologyIndian Institute of Technology MadrasChennaiIndia
  2. 2.201, Bhupat and Jyoti Mehta School of Biosciences, Department of BiotechnologyIndian Institute of Technology MadrasChennaiIndia

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