This protocol describes the isolation of primary adult mouse cardiomyocytes using sequential calcium depletion, enzymatic digestion, and gradual calcium reintroduction. The approach is adapted from the method of Matthew Ackers-Johnson et al.

Ahlers, B. A., Song, J., Wang, J., Zhang, X., Carl, L. L., Tadros, G. M., Rothblum, L.I., and Cheung, J. Y. (2005). Journal of Applied Physiology, 98(6), 2169-2176