Measuring Cardiomyocyte Contractility and Calcium Handling In Vitro

Methods Mol Biol. 2018;1816:93-104. doi: 10.1007/978-1-4939-8597-5_7.

Abstract

In vitro measurements of cardiomyocyte contractility and Ca2+ handling have been used as a platform for determining physiological consequence of various genetic manipulations and identifying potential therapeutic targets for the treatment of heart failure. The Myocyte Calcium and Contractility System (IonOptix) offers a simultaneous trace of sarcomere movements and changes of intracellular Ca2+ levels in a single cardiomyocyte. Herein, we describe a modified protocol for the isolation of adult cardiomyocytes from murine hearts and provide a step-by-step description on how to analyze cardiomyocyte Ca2+ transient and contractility data collected using the IonOptix system. In our modified protocol, we recommend a novel cannulation technique which simplifies this difficult method and leads to improved viability of isolated cardiomyocytes. In addition, a comprehensive analysis of intracellular Ca2+ handling, SR Ca2+ load, myofilament Ca2+ sensitivity, and cardiomyocyte contractility is described in order to provide important insights into myocardial mechanics.

Keywords: Calcium handling; Cardiomyocyte; Contractility; Heart disease; Intracellular calcium.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Calcium Signaling
  • Cardiac Catheterization / methods
  • Cell Culture Techniques / methods
  • Cell Separation / methods
  • Cells, Cultured
  • Mice
  • Myocardial Contraction*
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Myofibrils / metabolism
  • Sarcomeres / metabolism
  • Sarcoplasmic Reticulum / metabolism

Substances

  • Calcium

Grant support