Minimal contribution of IP₃R2 in cardiac differentiation and derived ventricular-like myocytes from human embryonic stem cells

Type 2 inositol 1,4,5-trisphosphate receptor (IP₃R2) regulates the intracellular Ca²⁺ release from endoplasmic reticulum in human embryonic stem cells (hESCs), cardiovascular progenitor cells (CVPCs), and mammalian cardiomyocytes. However, the role of IP₃R2 in human cardiac development is unknown and its function in mammalian cardiomyocytes is controversial. hESC-derived cardiomyocytes have unique merits in disease modeling, cell therapy, and drug screening. Therefore, understanding the role of IP₃R2 in the generation and function of human cardiomyocytes would be valuable for the application of hESC-derived cardiomyocytes. In the current study, we investigated the role of IP₃R2 in the differentiation of hESCs to cardiomyocytes and in the hESC-derived cardiomyocytes. By using IP₃R2 knockout (IP₃R2KO) hESCs, we showed that IP₃R2KO did not affect the self-renewal of hESCs as well as the differentiation ability of hESCs into CVPCs and cardiomyocytes. Furthermore, we demonstrated the ventricular-like myocyte characteristics of hESC-derived cardiomyocytes. Under the α₁-adrenergic stimulation by phenylephrine (10 μmol/L), the amplitude and maximum rate of depolarization of action potential (AP) were slightly affected in the IP₃R2KO hESC-derived cardiomyocytes at differentiation day 90, whereas the other parameters of APs and the Ca²⁺ transients did not show significant changes compared with these in the wide-type ones. These results demonstrate that IP₃R2 has minimal contribution to the differentiation and function of human cardiomyocytes derived from hESCs, thus provide the new knowledge to the function of IP₃R2 in the generation of human cardiac lineage cells and in the early cardiomyocytes.