Isolation, culturing, and characterization of cardiac muscle cells from nonhuman primate heart tissue.

Abstract

Cardiac safety pharmacology requires in vitro testing of all drug candidates before clinical trials in order to ensure they are screened for cardiotoxic effects which may result in severe arrhythmias and, ultimately, cardiomyopathy (Chi, Nat Rev Drug Discov 12:565-567, 2013). Given the physiological similarities between nonhuman primates and humans, isolated primate cardiac muscle cells are an ideal animal model for such in vitro testing. The aims of this chapter are to describe two methods for isolating and culturing primate cardiac muscle cells. One method uses mechanical dissociation of the tissue followed by placing the small pieces onto a Petri dish and culturing these tissue explants. The other method also uses mechanical dissociation but is then followed by enzymatic digestion and culturing of the cell suspension. Methods are also described for phenotypically characterizing cardiac muscle cells by flow cytometry. Based on the location within the heart tissue chosen for cell isolation, a dividing population of cardiac muscle cells expressing cardiomyocyte cell markers was obtained.