Priming adult human hepatocytes as organoids for liver tissue engineering

While human hepatocytes in the adult liver perform hundreds of life-sustaining functions, culture methods to support their phenotype and function in vitro remain elusive. Here, we describe establishment of an organoid culture system for adult human hepatocytes and demonstrate its robustness by generating organoids from eight different adult donors. Organoids accurately model the phenotype and morphology of hepatocytes in the adult liver, express a mature liver transcriptome distinct from fetal hepatocytes, and exhibit mature functions such as inducible cytochrome activity. Finally, hepatocyte organoids efficiently engraft in vivo in engineered tissues, exhibiting up to 25-fold greater functionality after engraftment compared to previous adult hepatocyte models. Furthermore, organoids negate the need for inclusion of exogenous non-parenchymal cells in engineered tissues to support engraftment, an important translational milestone. These results demonstrate the potential of adult human hepatocyte organoids for basic and translational applications such as pharmaceutical screening and hepatic regenerative medicine. Adult human hepatocytes organoids were grown, digested to single cells after 5 or 15 days of growth, and analyzed using scRNAseq. ----------------------------------------- Authors state "Unfortunately the raw data files were lost when our server crashed recently."