Development of rat liver 3D organoid methods to address genotoxicity screening

Project Summary Studying cells in simple 2D culture systems has many advantages with of ease of use, ability to screen many conditions in a short amount of time, targeted addition or deletion of genes, proteins or other components and focused study of specific pathways representing only a few. However, that simplicity can also lead to loss of key functions and an overall lack of relevancy to whole organisms, especially humans. In an effort to bridge this gap between cells in a dish and a human being, in vitro models are being developed that mimic the function of whole organs, so-called organotypic cultures. These models represent an opportunity to study highly complex, multicellular systems that will respond more like actual organs as opposed to cells grown in a simple monolayer. Our studies will develop a model of rat liver cells grown in a 3-dimensional format and pair that with an efficient, high-content analytical platform that will provide information on DNA function and cell health. The value of these types of models is significant for both basic science and more targeted research efforts such as human safety studies. Assessment of the safety of compounds like drug candidates, industrial chemicals and consumer products relies on preclinical test models to report useful information of human safety. The ability of cell-based models to do this is often challenged when the in vitro and in vivo systems lack sufficient relevancy to the human condition. The proposed 3D organ models may overcome existing limitations and result in lower costs while providing better information. The combination of data-rich assays that enable fast and efficient assessment of in vitro animal-derived organ models represents an opportunity to improve safety testing and create tools that will benefit numerous other research initiatives that rely on in vitro systems.Project Narrative Recent advances in cell culture have lead to the development of highly complex miniaturized tissue systems that replicate the function of human organs. The use of more relevant animal-derived organ models and the development of tools to better assess their responses could greatly improve chemical/drug safety studies, as well as aid in discoveries made in the basic sciences. This project seeks to create novel assays that can be used to provide quantitative information on activity and toxicological responses of complex 3D liver models to improve the relevancy and efficacy of these important in vitro organ systems.