Isolation and Culture of Neonatal Mouse Cardiomyocytes

The overall goal of this procedure is to isolate and culture neonatal mouse cardiomyocytes for use in subsequent assays. This is accomplished by first dissecting the hearts of neonatal mice, and then manually cutting the heart tissue into small fragments. In the second step, the cardiac cells are dissociated through a series of enzymatic digestions. Next, the isolated cells are plated, to enrich for cardiomyocytes. Finally, the cultured cardiomyocytes can be observed, adhering to the plate, and displaying spontaneous beating behavior. Ultimately, the subcellular localization of proteins within cardiac muscle cells transfected with eukaryotic expression constructs, can be observed by immunofluorescent microscopy.

- The main advantage of this technique over other methods is the robustness and reproducibility in the isolation and culture of neonatal mouse cardiomyocytes.