An improved method for isolation and culture of neonatal rat cardiomyocytes

Objective To explore a simple and effective primary culture method of neonatal rat cardiomyocytes. Methods The ventricular myocardium was removed from neonatal rat within 24 h after birth. The cardiomyocytes were separated by repeated digestion with 0. 08% trypsin and collected by centrifugation. Differential adhesion method and 5-bromodeoxyuridine( Brd U) were used in purifying cardiomyocytes. The action potential was measured by whole cell patch-clamp. The calcium transients and mitochondrial membrane potential of cardiomyocytes were observed by confocal microscope. Results The cell viability was more than 93%. The purity of cardiomyocytes was more than 96%. The morphology of cardiomyocytes appeared normal. The calcium transients were steady and regular. The mitochondrial membrane potential and action potential of cardiomyocytes were normal. Conclusion The primary culture method of neonatal rat cardiomyocytes is simple and effective,which could be applied to different cardiology researches.