Generation and characterization of mature hepatocyte organoids for liver metabolic studies

The liver plays a central role in whole body metabolic regulation. Studies of liver metabolism are mostly done in vivo in animal models due to the lack of reliable in vitro systems that can recapitulate liver metabolic functions and their regulation. Hepatocyte organoids (HOs) generated in vitro recently are powerful tools for liver regeneration. Here we developed a novel culture condition that enabled successful generation of mature hepatocyte organoids (MHOs) with metabolic functions characteristic of adult livers. We showed that the MHOs can be used to study gene expression that exhibit zonal patterns in vivo, hepatic proliferation during tumorigenesis and metabolic alterations in disorders of both alcoholic and non-alcoholic fatty liver diseases. To induce hepatic maturation of HOs, several culture conditions were tested in the experiments. The HOs were switched to the maturation medium for HO maturation. Samples were collected on day 3, day 6, and day 12 after the medium switch. Each sample group has three replicate samples.