Phenotypic Cell-Based Screening of a High Content Imaging Cell Health Assay using the IN Cell Analyzer 2000

Cell health assays play a key role in early drug discovery as they provide a crucial means of ranking novel chemical or biotherapeutic leads during screening and help to gain a better understanding of changes at the cellular level in disease pathologies. Recently there has been the emergence of High Content Screening (HCS), which integrates cell based imaging with multiple readouts in relevant cells in a high-throughput format. This has allowed in vitro cell health assays to be developed into High Content Imaging assays and used for HCS where they employ a multi-parametric approach enabling the capture of both phenotypic and mechanistic information of the cells. High Content Screening of cell health assays is now commonly being used as an early predictive toxicity assay to evaluate compound toxicity, in particular hepatotoxicity.
We demonstrate how we established an in-house cell health assay to be used for high content phenotypic screening using the IN Cell Analyzer 2000 with the aim to screen subsets of MRCT compound libraries and in-house compounds. The assay development process is illustrated using drugs withdrawn from the market or known to cause hepatotoxicity as tool compounds and initial screening of the FDA set in HepG2 cells to validate the assay. Genedata Screener analysis software was used for data processing and to visualise the data generated from the high content imaging screen.