Enrichment of Cardiomyocytes in Primary Cultures of Murine Neonatal Hearts

  • Sreejit Parameswaran
  • Rajalakshmi Santhakumar
  • Prasanna Vidyasekar
  • Rama S. Verma
Part of the Methods in Molecular Biology book series (MIMB, volume 1299)

Abstract

In vitro culture of neonatal murine cardiomyocytes is vital for understanding the functions of the heart. Cardiomyocyte cultures are difficult to maintain because they do not proliferate after birth. The maintenance of primary cultures of viable and functional cardiomyocytes is considerably affected by the yield from initial steps of isolation procedures. This protocol describes an efficient and rapid method for isolation and maintenance of long-term cultures of neonatal murine cardiomyocytes by effectively shortening the trypsin enzyme digestion period and the cardiomyocyte enrichment step.

Key words

Primary cell culture Neonatal mice Murine cardiomyocyte enrichment Immunocytochemistry 

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).

References

  1. 1.
    Yamashita N, Nishida M, Hoshida S, Kuzuya T, Hori M, Taniguchi N et al (1994) Induction of manganese superoxide dismutase in rat cardiac myocytes increases tolerance to hypoxia 24 h after preconditioning. J Clin Invest 94:2193–2199CrossRefPubMedCentralPubMedGoogle Scholar
  2. 2.
    Bahi N, Zhang J, Llovera M, Ballester M, Comella JX, Sanchis D (2006) Switch from caspase-dependent to caspase-independent death during heart development: essential role of endonuclease G in ischemia-induced DNA processing of differentiated cardiomyocytes. J Biol Chem 281(32):22943–22952CrossRefPubMedGoogle Scholar
  3. 3.
    Limaye DA, Shaikh ZA (1999) Cytotoxicity of cadmium and characteristics of its transport in cardiomyocytes. Toxicol Appl Pharmacol 154(1):59–66CrossRefPubMedGoogle Scholar
  4. 4.
    Wang GW, Kang YJ (1999) Inhibition of doxorubicin toxicity in cultured neonatal mouse cardiomyocytes with elevated metallothionein levels. J Pharmacol Exp Ther 288(3):938–944PubMedGoogle Scholar
  5. 5.
    Kruppenbacher JP, May T, Eggers HJ, Piper HM (1993) Cardiomyocytes of adult mice in long-term culture. Naturwissenschaften 80:132–134CrossRefPubMedGoogle Scholar
  6. 6.
    Chlopclkova Š, Psotova J, Miketova P (2001) Neonatal rat cardiomyocytes—a model for the study of morphological, biochemical and electrophysiological characteristic of the heart. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 145:49–55CrossRefGoogle Scholar
  7. 7.
    Nuss HB, Marban E (1994) Electrophysiological properties of neonatal mouse cardiac myocytes in primary culture. J Physiol 479(2):265–279CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Sreejit Parameswaran
    • 1
  • Rajalakshmi Santhakumar
    • 1
  • Prasanna Vidyasekar
    • 1
  • Rama S. Verma
    • 1
  1. 1.Stem Cell and Molecular Biology Laboratory, Department of BiotechnologyIndian Institute of Technology MadrasChennaiIndia

Personalised recommendations