Techniques for differentiating human induced pluripotent stem cells (hiPSCs) into cardiomyocytes (hiPSC-CMs) can yield up to 98% purity, yet control over hiPSC-CM fate is lacking. Here, we present a protocol for insulin dosing during the expansion and maturation of hiPSC-derived cardiomyocytes. We describe steps for differentiation, purification, replating, cryopreservation, and thawing. We then detail a procedure for expansion using insulin and Wnt/β-catenin signaling, followed by steps to initiate maturation by either removing insulin completely or adding a low dose of insulin. For complete details on the use and execution of this protocol, please refer to Yuan et al.
Verbueken et al. (Mon,) studied this question.