Isolation and culture of adult mouse cardiac myocytes

Timothy D. O'Connell, Manoj C. Rodrigo, Paul C. Simpson

Research output: Chapter in Book/Report/Conference proceedingChapter

201 Scopus citations

Abstract

Cardiac myocytes are activated by hormonal and mechanical signals and respond in a variety of ways, from, altering contractile function to inducing cardio-protection and growth responses. The use of genetic mouse models allows one to examine the role of cardiac-specific and other genes in cardiac function, hypertrophy, cardio-protection, and diseases such as ischemia and heart failure. However, studies at the cellular level have been, hampered by a lack of suitable techniques for isolating and culturing calcium-tolerant, adult mouse cardiac myocytes. We have developed a straightforward, reproducible protocol for isolating and culturing large numbers of adult mouse cardiac myocytes. This protocol is based on the traditional approach of retrograde perfusion of collagenase through the coronary arteries to digest the extracellular matrix of the heart and release rod-shaped myocytes. However, we have made modifications that are essential for isolating calcium-tolerant, rod-shaped adult mouse cardiac myocytes and maintaining them in culture. This protocol yields freshly isolated adult mouse myocytes that are suitable for biochemical assays and for measuring contractile function and calcium, transients, and cultured myocytes that are suitable for most biochemical and signaling assays, as well as gene transduction using adenovirus.

Original languageEnglish (US)
Title of host publicationCardiovascular Proteomics
Subtitle of host publicationMethods and Protocols
EditorsFernando Vivanco
Pages271-296
Number of pages26
DOIs
StatePublished - Feb 5 2007

Publication series

NameMethods in Molecular Biology
Volume357
ISSN (Print)1064-3745

Keywords

  • Adenovirus
  • Apoptosis
  • Cardiac myocyte
  • Cell culture
  • Cell isolation
  • Cell signaling
  • Hypertrophy
  • β-adrenergic

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  • Cite this

    O'Connell, T. D., Rodrigo, M. C., & Simpson, P. C. (2007). Isolation and culture of adult mouse cardiac myocytes. In F. Vivanco (Ed.), Cardiovascular Proteomics: Methods and Protocols (pp. 271-296). (Methods in Molecular Biology; Vol. 357). https://doi.org/10.1385/1-59745-214-9:271