Isolation of cardiac myocytes and mesenchymal progenitor cells (MPCs)

To isolate cardiac myocytes explanted hearts were perfused with a Ca²⁺ free perfusion buffer and subjected to digestion in a collagenase buffer. Myocytes were dissociated and gravity precipitated in the presence of 200 mM ATP and centrifugation at 20 g. The isolated myocytes were re-introduced to increasing concentrations of calcium at a final concentration of 1.5 mM of CaCl2 and placed in culture dishes or cover glasses coated with laminin.

Cardiac MPCs were isolated by sorting of non-myocyte fraction of cardiac cells against anti CD44 and anti platelet-derived growth factor α (PDGFRA) antibodies, per published protocols ^34-36.