Abstract

Organoids For Disease Modeling and In Vitro Assessment of Drug Effects by High-Content Imaging and 3D Analysis

Oksana Sirenko, Sr. Imaging Applications Scientist, Molecular Devices

3D cell models representing different tissues were successfully used for studying complex biological effects, tissue architecture, and functionality. While complexity of 3D models remains a hurdle for the wider adoption in research and drug screening, significant progress has been made in developing tools to automate the culture, monitoring, and analysis of these structures. This enabled an increase in quality of information, throughput, and precision of complex biological assays. We describe automated cell culture and high-content imaging methods that allow monitoring and characterization of growth and differentiation of lung organoids, as well as toxicity and drug effects. Increase in size and complexity of developing organoids was monitored during eight weeks of development. We demonstrated concentration-dependent toxicity effects of several drugs that have been known to cause lung damage. In addition, responses to several anti-cancer drugs were monitored in 3D tumoroids derived from primary tumor tissues.  Advanced image analysis allowed 3D reconstitution and complex analysis of organoid structures, characterization of cell morphology and viability, as well as determining expression level of differentiation markers. Image analysis included traditional and Machine Learning-based tools to increase the ability to discern important biological information from the complex multi-dimensional datasets. These quantitative characterization of organoids morphology can be used for studying disease phenotypes and compound effects in complex assay models.