In the past few years, several different methods for differentiation of directed cardiomyocytes from human pluripotent stem cells (hPSCs) in chemically defined conditions have been reported, including our own (Burridge et al., 2014; Lian et al., 2012; Lin et al., 2017). To help researchers adapt to our simple and cost-effective method, we provide step-by-step protocols for hPSC-cardiomyocyte differentiation, including hPSC culture, cardiomyocyte differentiation, cardiomyocyte passaging, and cryopreservation. For complete details on the use and execution of this protocol, please refer to Lin et al. (2017).