3D Spheroidal Human Liver Models for Drug Toxicity Testing
thesisposted on 2020-12-01, 00:00 authored by Shivam Ohri
Drug-induced liver injury (DILI) remains the leading cause of preclinical and clinical drug attrition, drug withdrawals from the market, and black-box warnings on currently marketed drugs. This study aims to provide an efficient model to predict drug-induced hepatotoxicity through optimization of a widely used three-dimensional liver model system, namely spheroids. Addressing the different rates of formation of monoculture spheroids, we evaluated the role of coculture with primary human hepatocytes by adding relevant cell types demonstrating differences in functionality and spheroid formation. We also assessed drug sensitivity of all the culture conditions using known toxic and non-toxic pharmaceutical drugs, providing a comprehensive comparison between different types of spheroids. Our results suggest that different monoculture spheroids formation rates can be mitigated using a coculture model system with few limitations related to drug sensitivity. Lastly, we fabricated microwells using poly (ethylene glycol) diacrylate (PEGDA), which is considered a cytocompatible and easy to use polymer, on a glass substrate with the aim of multiple spheroid formation per well in a 96 well plate and enabling full media changes that reduces handling time of the cultures. The results suggest consistent spheroid formation with a detectable level of functions for all culture conditions, enabling future use of the model system for many applications, including drug screening.