Resveratrol effectively promoted hepatocyte differentiation of hESCs by enhancing mitochondrial function through PGC-1α PPARγ pathway.

Despite their significant potential, hepatocytes derived from human embryonic stem cells (hESCs) exhibits characteristics of immature hepatocytes, similar to fetal liver cells. The quantity of the mitochondria, which functions as the main energy generators in mammalian cells, display the variation across distinct cell types with specific energy demands. However, the specific correlation between this variability and hepatocyte differentiation remains elusive. Recently, we developed a 3D suspension differentiation method to differentiate hESCs into hepatocyte spheres, and observed that resveratrol (RES) treatment further promoted hepatocyte maturation and improved the polarization of hepatocytes during hepatocyte differentiation of hESCs. We also found that the treatment with RES resulted in a significant augmentation in intracellular ATP content, mitochondrial DNA copy number, and protein subunits associated with mitochondrial biogenesis, indicating that mitochondrial function was enhanced. In addition, PGC-1α, a crucial regulator of mitochondrial biogenesis, was up-regulated during RES treatment, and eventually our further investigation revealed that RES effectively promoted hESC-derived hepatocyte maturation and polarization by enhancing mitochondrial function through PGC-1α-mediated activation of PPARγ pathway. Thus, our results demonstrated that RES-mediated enhancement of mitochondrial biogenesis could promote hepatocyte differentiation from hESCs. Therefore, this finding provides a new avenue to generate more mature hepatocytes for pharmaceutical and toxicological studies as well as for cell-based therapies.