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Briefly, the heart obtained from neonatal rat was immersed in oxygenated Tyrodes solution and ventricles were isolated from the heart under a stereomicroscope. Then these ventricular tissues were immersed in Ca 2+ -free Tyrodes solution at room temperature to remove extracellular Ca 2+. The tissues were digested by collagenase (5 mg/ml) for 60 min at 37°C.

The digested tissues were transported to 250 cc su ne kadar Kraftbrühe (KB) solution and pipetteing was gently performed to make clenbuterol norge a cell suspension. Isolated naonatal cardiomyocytes were seeded into culture plates and cultured overnight in DMEM supplemented with 10% fetal bovine serum (FBS).