Abstract
Human primary cardiomyocytes are a valuable in vitro model for studying human heart physiology and disease. Here, we present a protocol for the dissociation of adult human primary cardiomyocytes using a methylcellulose-supplemented digestion solution. We describe steps for sample collection, tissue pre-processing, and cell dissociation. We then detail procedures for calcium reintroduction and viability assessment. This protocol can significantly improve the efficiency of cardiomyocyte isolation and subsequent cell function, making them an even more reliable model for cardiac research. For complete details on the use and execution of this protocol, please refer to Shi et al.1.