Protocol for primary cardiac cell culture from individual neonatal mice

Here, we present an adapted primary cardiac cell culture protocol (from Ehler et al.¹) to generate cardiac cultures from single neonatal mouse hearts that retain cardiomyocytes together with endogenous fibroblast and endothelial cell populations. The production of cultures from individual animals enables genotype- and sex-specific analysis. We describe the optimized enzymatic dissociation of an individual heart and culture conditions that support cardiomyocyte maintenance. The sex of each sample is determined using a previously established PCR-based genotyping assay.².