Directed Differentiation of iPSCs to Cardiomyocytes

Human pluripotent stem cells were differentiated into iCMs using an established differentiation procedure as previously described (Wanjare et al., 2017 (link)). Briefly, iCMs were generated using a directed differentiation method that involved incubating iPSCs with Wnt agonist CHIR 99021 (6 μM, Selleck), followed by Wnt antagonist IWR-1 (5 μM, Selleck) in Roswell Park Memorial Institute (RPMI) 1640 medium (Thermo Fisher), supplemented with insulin-free B27 (Invitrogen). After 7 days, the culture medium was replaced with RPMI 1640 supplemented with B27 and insulin (Invitrogen). To enrich the iCM populations, the differentiating cells were deprived of glucose after 9 days. The iCMs were used for experiments after 15 days of differentiation. Only dishes containing >80% spontaneously contracting iCMs were used for the described experiments.