Abstract

Automation of 3D Organoid Culture with Monitoring, Imaging and Analysis of Complex Biological Effects

Oksana Sirenko, Sr. Imaging Applications Scientist, Molecular Devices

There is an unmet need for cell-based model systems that better resemble human biology to improve pre-clinical selection of drug candidates. 3D cell models representing various 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 for automated cell culture, monitoring, and image analysis. We will describe an integrated cell culture and a high-content imaging process that allows automated culture, monitoring, maintenance, and characterization of growth and differentiation of organoids and stem cells, as well as testing the effects of various compounds. We will discuss the methods for automation of three complex workflows: maintenance and monitoring of iPSC cell culture, culturing and testing toxicity effects in lung organoids, and monitoring of the development of intestinal organoids. The automated integrated system included ImageXpress Confocal HT.ai High-Content Imaging System, (IXM-C HT.ai confocal imaging system), automated CO₂ incubator (Wave STX44), a liquid handler (Biomek i7), as well as an automation robotic system and software. Described workflow demonstrates the useful tools for increased throughput and automation in organoid assays and compound screening, and analysis methods and descriptors for gaining more information from complex assays ?useful for studying disease phenotypes and compound effects.