Isolation and in vitroculture of primary cardiomyocytes from adult zebrafish hearts

This protocol describes how to isolate primary cardiomyocytes from adult zebrafish hearts and culture them for up to 4 weeks, thereby using them as an alternative to in vivoexperiments. after collagenase digestion of the ventricle, cells are exposed to increasing calcium concentrations in order to obtain high-purity cardiomyocytes. the whole isolation process can be accomplished in 4–5 h. the culture conditions we established allow the cells to preserve their mature sarcomeric integrity and contractile properties. Furthermore, adult zebrafish cardiomyocytes in culture, similarly to zebrafish in vivoheart regeneration, undergo partial dedifferentiation and, in contrast to their mammalian counterparts, are able to proliferate. our protocol enables the study of structural and functional properties in close-to-native cardiomyocytes and allows the application of in vitrotechniques and assays that are not feasible to perform in living animals.