Human ESC-derived Expandable Hepatic Organoids Enable Therapeutic Liver Repopulation and Pathophysiological Modeling of Alcoholic Liver Injury

Abstract

Introduction: Here, we report the generation of human ESC-derived, expandable hepatic organoids (hEHOs) enabling therapeutic liver repopulation and pathophysiological modeling of alcoholic liver injury. Methods: The culture system including a mix of A83-01, Forskolin, R-spondin 1, Wnt3a and EGF along with the extracellular matrix was used to generate hEHOs. The liver repopulation potential of hEHOs was assayed in FRG mouse model of inherited metabolic liver disorders. The tumorigenesis and lineage restriction of hEHOs were evaluated by transplanting the hEHOs into epididymal fat pads of NOD/SCID mice. To model alcoholic liver disease, a derivative model by incorporating human fetal liver mesenchymal cells (hFLMCs) into the hEHOs, referred to as hFLMC/hEHO was generated and treated with 100mM ethanol for 7days. Results: The hEHOs stably maintain phenotypic features of bipotential liver stem/progenitor cells that can differentiate into functional hepatocytes or cholangiocytes. The hEHOs can expand for 20 passages enabling large scale expansion to cell numbers requisite for industry or clinical programs. The hEHOs display remarkable repopulation capacity in injured livers of FRG mice following transplantation. If implanted into the epididymal fat pads of immune-deficient mice, they do not generate non-hepatic lineages and have no tendency to form teratomas. The hFLMC/hEHO can model alcoholic liver disease-associated pathophysiologic changes, including oxidative stress generation, steatosis, inflammatory mediators release and fibrosis, under ethanol treatment. Conclusion: The hEHOs have considerable potential to be a novel, ex vivo pathophysiological model for studying alcoholic liver disease as well as a promising cellular source for treating human liver diseases.