A roadmap for the characterization of energy metabolism in human cardiomyocytes derived from induced pluripotent stem cells

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are an increasingly employed model in cardiac research and drug discovery. As cellular metabolism plays an integral role in determining phenotype, the characterization of the metabolic profile of hiPSC-CM during maturation is crucial for their translational application. In this study we employ a combination of methods including extracellular flux, C-13-glucose enrichment and targeted metabolomics to characterize the metabolic profile of hiPSC-CM during their matura-tion in culture from 6 weeks, up to 12 weeks. Results show a progressive remodeling of pathways involved energy metabolism and substrate utilization along with an increase in sarcomere regularity. The oxidative ca-pacity of hiPSC-CM and particularly their ability to utilize fatty acids increased with time. In parallel, relative glucose oxidation was reduced while glutamine oxidation was maintained at similar levels. There was also ev-idence of increased coupling of glycolysis to mitochondrial respiration, and away from glycolytic branch path-ways at later stages of maturation. The rate of glycolysis as assessed by lactate production was maintained at both stages but with significant alterations in proximal glycolytic enzymes such as hexokinase and phosphofructo-kinase. We observed a progressive maturation of mitochondrial oxidative capacity at comparable levels mitochondrial content between these time-points with enhancement of mitochondrial network structure. These results show that the metabolic profile of hiPSC-CM is progressively restructured, recapitulating aspects of early post-natal heart development. This would be particularly important to consider when employing these cell model in studies where metabolism plays an important role.