Abstract
Drug-induced liver injury is one of the leading causes of market withdrawal in the pharmaceutical industry and poses a serious health risk to affected patients. Here, we developed a 3D in vitro model of the human liver, the OrganoPlate LiverTox compatible with automated liquid handling and validated for hepatotoxicity screening. To build the model, up to 96 independent 3D perfused cultures were established on MIMETAS’ OrganoPlate 2-lane. Induced pluripotent stem cell-derived hepatocytes (iHep) in extracellular matrix were added to a microfluidic channel, following which endothelial and Kupffer cells were added to an adjacent channel. Characterization of the model revealed hepatocyte function including CYP3A4 activity and albumin production for up to 14 days. Fetal hepatocyte marker alpha-fetoprotein (AFP) declined over the 14 day culture, supporting iHep maturation. Assay validation studies using troglitazone as a positive hepatotoxic control revealed robust Z-factors ≥ 0.2 for albumin, urea, iHep viability (propidium iodide staining), and iHep nuclear size (Hoechst 33342 staining) assay readouts. Using these assays, 159 compounds of known hepatotoxicity were screened in the OrganoPlate LiverTox (50 μM, 72 h) and ranked by a composite score by combining the assay readouts. A follow-up dose response evaluation of select hits suggested the albumin assay to be the most sensitive readout in calculating TC50 values. Together, the OrganoPlate LiverTox is a promising platform for hepatotoxicity detection and has the potential to be used in a high throughput screening capacity.
