3D human hepatic organoids for testing Fibrosis, Cholestasis and Phospholipidosis

Current in vitro models of liver fibrosis consist in simple mono-layer cultures of rodent hepatic stellate cells (HSC), thereby ignoring the role of hepatocyte injury. We developed a human 3D hepatic organoid culture model including HepaRG cells and primary human hepatic stellate cells (HSCs), for the screening of pro-fibrotic compounds. These scaffold-free organoids were characterized for CYP induction, albumin secretion, and hepatocyte and HSC-specific gene expression. The metabolic competence of organoids over 21 days allows activation of HSCs in a drug- and hepatocyte-dependent manner. A range of compounds including Acetaminophen, Methotrexate, Allyl Alcohol, Pyperonyl Butoxide, Bosentan, Valproic Acid and Amiodarone has been tested. The model has been successfully used for the detection of various drug induced liver injury (DILI) mechanisms including fibrosis, cholestasis and phospholipidosis both after single and repeated exposures. In depth characterization of the organoids is ongoing as well as the use of rat and mouse primary cells. This novel hepatic organoid culture model is the first to detect hepatocyte-dependent and compound-induced HSC activation, thereby representing an important step towards in vitro testing for drug-induced liver injury especially fibrosis