Functional culture and in vitro genetic and small-molecule manipulation of adult mouse cardiomyocytes

Primary adult cardiomyocyte (aCM) represent the mature form of myocytes found in the adult heart. However, culture of aCMs in particular is challenged by poor survival and loss of phenotype, rendering extended in vitro experiments unfeasible. Here, we establish murine aCM culture methods that enhance survival and maintain sarcomeric structure and Ca2+ cycling to enable physiologically relevant contractile force measurements. We also demonstrate genetic and small-molecule manipulations that probe mechanisms underlying myocyte functional performance. Together, these refinements to aCM culture present a toolbox with which to advance our understanding of myocardial physiology. Callaghan et al. present a combinatory approach to culturing harvested adult mouse cardiomyocytes (aCMs). Under traditional culture protocols, aCMs rapidly lose their phenotype and undergo cell death. With their protocol, the authors show aCMs remain viable and retain their phenotype for 7 days, enough time to do genetic manipulation and small molecule screening.