Multi-cell type human liver microtissues for hepatotoxicity testing

TLDR

Traditional 2‑D hepatic cultures lose hepatocyte phenotype and fail to predict drug‑induced liver injury, while 3‑D systems that preserve polarization and cell‑cell contacts in a multi‑cell environment are needed for accurate in‑vitro hepatotoxicity assessment. The authors aim to develop a 3‑D, multi‑cell type liver microtissue model from primary human hepatocytes and non‑parenchymal cells for routine 96‑well toxicity screening that reduces the risk of clinical hepatotoxicity. They generate stable microtissues that maintain hepatocyte phenotype for five weeks, enabling long‑term testing of acetaminophen and diclofenac and allowing inflammatory responses via Kupffer cells to be assessed. The microtissues remain functional for five weeks, support long‑term toxicity assays, and Kupffer cells respond to LPS, demonstrating the model’s capacity to detect both drug‑induced and inflammation‑mediated hepatotoxicity, as shown with trovafloxacin.

Abstract

Current 2-dimensional hepatic model systems often fail to predict chemically induced hepatotoxicity due to the loss of a hepatocyte-specific phenotype in culture. For more predictive in vitro models, hepatocytes have to be maintained in a 3-dimensional environment that allows for polarization and cell-cell contacts. Preferably, the model will reflect an in vivo-like multi-cell type environment necessary for liver-like responses. Here, we report the characterization of a multi-cell type microtissue model, generated from primary human hepatocytes and liver-derived non-parenchymal cells. Liver microtissues were stable and functional for 5 weeks in culture enabling, for example, long-term toxicity testing of acetaminophen and diclofenac. In addition, Kupffer cells were responsive to inflammatory stimuli such as LPS demonstrating the possibility to detect inflammation-mediated toxicity as exemplified by the drug trovafloxacin. Herewith, we present a novel 3D liver model for routine testing in 96-well format capable of reducing the risk of unwanted toxic effects in the clinic.