An Efficient and Convenient Method for Isolation and Culturing of Neonatal Rat Cardiomyocytes.

Abstract

For isolation of neonatal rat cardiomyocytes (NRCM) the ventricular muscles of neonatal rats were treated with different digestive solutions: 0.06% trypsin (method I), 0.08% collagenase II (method II), 0.06% trypsin and 0.08% collagenase II for stepwise digestion (methods III and IV). After enzymatic dissociation of the tissue, the complete medium was added to stop this process. The cells suspensions obtained by methods I-III were collected and centrifuged. In contrast, the novel and improved method IV did not use centrifugation. Instead, various methods of adhesion were employed to separate non-myocardial cells. The isolation methods were compared by the quantity, survival rate, morphology, spontaneous pulsation rate, purity, and vitality of NRCM. These assessments showed that isolation method IV is a simple, efficient, and convenient way to obtain NRCM for culturing.