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Isolation and Culture of Adult Mouse Cardiac Myocytes

  • Timothy D. O’Connell
  • Manoj C. Rodrigo
  • Paul C. Simpson
Part of the Methods in Molecular Biology™ book series (MIMB, volume 357)

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.

Key Words

Cardiac myocyte cell isolation cell culture hypertrophy cell signaling apoptosis adenovirus β-adrenergic 

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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Timothy D. O’Connell
    • 1
    • 2
    • 3
  • Manoj C. Rodrigo
    • 4
    • 5
    • 6
  • Paul C. Simpson
    • 4
    • 5
    • 6
  1. 1.Cardiovascular Research InstituteThe University of South DakotaSioux Falls
  2. 2.Department of MedicineThe University of South DakotaSioux Falls
  3. 3.South Dakota Health Research FoundationThe University of South DakotaSioux Falls
  4. 4.Cardiology DivisionSan Francisco VA Medical CenterSan Francisco
  5. 5.Department of MedicineThe University of California at San FranciscoSan Francisco
  6. 6.Cardiovascular Research InstituteThe University of California at San FranciscoSan Francisco

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