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H9c2 cells were induced to cardiomyocyte differentiation through reduction of FBS content from 10 to 1% in the cell culture medium and addition of retinoic acid (RA) for 6 days. 106 cells were plated and, once they reached approximately 80% confluence, differentiation was initiated by changing the medium to high-glucose DMEM (same supplementation) but with 1% FBS and 10 nmol/L retinoic acid (RA), resulting in an increase in cardiac-like cells. The differentiation medium was changed every 2 days (Pereira et al., 2011; Suhaeri et al., 2015; Patten et al., 2017).
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