Objective

Drug-induced hepatotoxicity is a major cause of drug failure during development and contributes to high rates of late stage drug attrition. Improving drug development outcomes remains challenging, due to a lack of robust assays and model systems to facilitate early and accurate prediction of hepatotoxicity. Advances in predictive toxicology have benefited from high content imaging approaches that enable rapid detection and analysis of complex hepatotoxic mechanisms. We have utilised this technology to develop robust and versatile 2D and 3D assay formats for the detection of drug induced phospholipidosis, a major adverse contributor to hepatotoxicity. We highlight the development of a highly reproducible and sensitive LipidTOX Red detection assay using HepG2 cells for the screening of phospholipidosis inducers using the ImageXpress MicroConfocal® imaging platform. Additionally, we demonstrate the adaption of this assay to 3D HepG2 spheroid cultures with additional multiplex detection of cellular viability and reactive oxidative species (ROS) markers. Together, these assays provide highly predictive solutions, adaptable for primary hepatocyte culture, to enable early detection of in vitro drug induced hepatotoxicity.