A 3D human liver model of nonalcoholic steatohepatitis

Abstract

We developed a 3D human liver model which exhibits many features of nonalcoholic steatohepatitis and that could become a platform for medium throughput drug screening. ABSTRACTWe have developed an in vitro preclinical 3D Non-Alcoholic SteatoHepatitis (NASH) model by co-culturing four human primary liver cell types: hepatocytes, stellate, endothelial and Kupffer cells. Cells were embedded in a hydrogel of rat collagen in 96-well plate and a NASH-like environment was induced with a medium containing free fatty acids (FFAs) and tumor necrosis factor  (TNF). This model was characterized by biochemical, imaging and transcriptomics analysis. On the one hand, we succeed in defining suitable culture conditions to maintain the 3D co-culture up to 10 days in vitro with the lowest level of steatosis, and reproducible low levels of inflammation and fibrosis. On the other hand, we induced NASH disease with a custom medium mimicking NASH features (hepatocyte injury, steatosis, inflammation and fibrosis). The 10-day cell viability and cost effectiveness of the model make it suitable for medium throughput drug screening and provide attractive avenues to better understand disease physiology and to identify and characterize new drug targets.

A 3D primary human cell-based in vitro model of non-alcoholic steatohepatitis for efficacy testing of clinical drug candidates

“…In contrast to studies presented here, the authors reported only low level of steatosis, inflammation and early fibrosis. No increased extracellular collagen fibers deposition and formation was demonstrated on histological level [51].…”

“…The gene expression of a range of cytokines/chemokines such as interleukin-1 (IL-1), chemokine (C-X-C motif) ligand 6 (CXCL6), C-C motif chemokine ligand 20 (CCL20), was increased by the loading of cells with FFAs and LPS (Supplementary figure 2d). Furthermore, the fibrosis biomarkers such as tissue inhibitor of metalloproteinases 1 (TIMP-1), collagens, fibronectin and elastin (Supplementary Figure 2d) were upregulated on gene expression level [2,51].…”

Obesity III: Obesogen assays: Limitations, strengths, and new directions

“…Essential factors such as spheroid size, time in culture, and culture media composition affect basal levels of xenobiotic metabolism and liver enzyme inducibility via activators of hepatic receptors such as the aryl hydrocarbon receptor (AhR), constitutive androstane receptor (CAR), and pregnane X receptor (PXR) [124]. Various co-culture techniques have also been developed for liver cell assays to recreate more tissue or disease-relevant environments for the evaluation of disease biology and toxicology [125].…”