Lipid metabolism remodeling in human cardiomyocyte differentiation and maturation.

Cardiac development involves extensive metabolic remodeling, yet dynamic changes of endogenous lipid species during human cardiomyocyte (CM) differentiation remain poorly characterized. In this study, we systematically performed high-coverage lipidomic profiling across five consecutive CM differentiation stages, using high-performance liquid chromatography. MYL2 expression confirmed ventricular lineage specification in late-stage CMs. Ceramide levels peaked at the cardiac progenitor cell (CPC) stage, suggesting a stage-specific association with the mesendoderm (MES)-to-CPC transition. Meanwhile, lipid categories such as monosialodihexosylganglioside, phosphatidylcholine (PC), lysophosphatidylcholine, and lysophosphatidic acid were markedly elevated in mature CMs. Integrated transcriptomic and chromatin accessibility analyses further delineated the temporal regulatory landscape underlying stage-dependent lipid remodeling during cardiogenic differentiation. Functional assays revealed that PC promotes the expression of mature CM marker genes and improves CM viability. Collectively, these findings establish a comprehensive, high-resolution dynamic lipidomic atlas of in vitro hESC-derived CM differentiation, providing a valuable resource for understanding lipid-associated mechanisms in heart development.