Electrotonic coupled metabolic purification of chick cardiomyocytes

Methods Mol Biol. 2015:1299:167-75. doi: 10.1007/978-1-4939-2572-8_14.

Abstract

Cardiomyocytes isolated from chick and rodent are widely used in studying cardiac physiology. However, contaminating non-cardiomyocytes are an inherent problem that hinders downstream analysis. Here, we report a novel electrical stimulation coupled with metabolic selection method using cytosine arabinoside (AraC) to efficiently eliminate contaminating cells in isolating chick embryonic cardiomyocytes. Compared with conventional methods of pre-plating or AraC alone, electrical stimulation coupled with AraC increased the percentage purity of cardiomyocytes by 2-6-fold with added effect of improved contractile function and maturation. This simple method could be useful in isolating and maintaining purified cardiomyocytes for long-term studies of cardiac physiology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cell Separation / methods*
  • Cells, Cultured
  • Chick Embryo
  • Chickens
  • Electric Stimulation
  • Fluorescent Antibody Technique
  • Microscopy, Confocal
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism*