Isolation of Cardiomyocyte and Nonc

Isolation of Cardiomyocyte and Noncardiomyocyte Fractions for Iron Quantitation.
Following cardiac perfusion, hearts were dissected into small pieces in ice-cold Hanks buffer and were subjected to collagenase P digestion at 37 °C for 1 h (11213857001; Roche Diagnostics). Following lysis of red blood cells, cell suspensions were passed through a 70-um sieve before being labeled using a cardiomyocyte isolation kit (130-100-825; Miltenyi Biotec). Separation of cardiomyocyte and noncardiomyocyte fractions was carried out using a MACS magnetic separation system (Miltenyi Biotec) according to the manufacturer’s instructions. Fractions were lysed immediately for ICP-MS analysis.