A modified Langendorff-free method for isolation of cardiomyocytes from adult rat heart

Abstract

Single isolated cardiomyocytes that are free from connective tissue and endothelium are widely used to investigate cellular and molecular mechanisms of cardiac regulation in the healthy hart and pathology. In this study, we present a new method to obtain single myocytes from adult rat hearts for biophysical experiments. We compare cell morphometry and parameters of intercellular calcium transient and contraction of rat ventricular cells isolated by Langendorff-free injection method and traditional Langendirff-based perfusion technique. Using the injection protocol, we found the number of viable cardiac myocytes with good morphometric characteristic was higher compared with the Langendorff-protocol. In both protocols, the cells retained rod-shape and sarcomere striation. The absence of statistically significant differences in the morphometric characteristics, calcium handling and mechanical activity of cardiomyocytes isolated by two different methods indicated that a new injection method is a simple adequate alternative for cardiomyocyte isolation to the Langendorff-based perfusion technique.Single isolated cardiomyocytes that are free from connective tissue and endothelium are widely used to investigate cellular and molecular mechanisms of cardiac regulation in the healthy hart and pathology. In this study, we present a new method to obtain single myocytes from adult rat hearts for biophysical experiments. We compare cell morphometry and parameters of intercellular calcium transient and contraction of rat ventricular cells isolated by Langendorff-free injection method and traditional Langendirff-based perfusion technique. Using the injection protocol, we found the number of viable cardiac myocytes with good morphometric characteristic was higher compared with the Langendorff-protocol. In both protocols, the cells retained rod-shape and sarcomere striation. The absence of statistically significant differences in the morphometric characteristics, calcium handling and mechanical activity of cardiomyocytes isolated by two different methods indicated that a new injection method is a simple adequa...