Abstract
In recent years, the assembly and annotation of chemogenomic libraries have gained interest by the phenotypic screening community. Apart from basic annotations of the compound potency and selectivity, these compound libraries benefit in particular from annotation regarding the effect of the inhibitors on cellular viability to distinguish between on-target effects of a compound and unspecific cytotoxicity. Here, we provide a protocol to determine viability as a first determinant in compound quality control, using the Incucyte live-cell imaging system. The compounds are classified according to their calculated growth rate to determine a cytotoxic, cytostatic, or healthy outcome. All compounds affecting the growth rate can be further evaluated regarding their specific effects on cell health in a high-content live-cell multiplex assay, described in Chapter 5.
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Acknowledgement
The authors are grateful for support by the Structural Genomics Consortium (SGC), a registered charity (No: 1097737) that receives funds from Bayer AG, Boehringer Ingelheim, Bristol Myers Squibb, Genentech, Genome Canada through Ontario Genomics Institute, Janssen, Merck KGaA, Pfizer and Takeda and by the German Cancer Research Center DKTK and the Frankfurt Cancer Institute (FCI). This project has received funding from the Innovative Medicines Initiative 2 Joint Undertaking (JU) under grant agreement No 875510. The JU receives support from the European Union’s Horizon 2020 research and innovation program and EFPIA and Ontario Institute for Cancer Research, Royal Institution for the Advancement of Learning McGill University, Kungliga Tekniska Hoegskolan, Diamond Light Source Limited. Disclaimer: This communication reflects the views of the authors, and the JU is not liable for any use that may be made of the information contained herein. A.T. is supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – grant number 259130777 (SFB1177). We would also like to thank Dr. Alexandra Stolz and her team for assistance with experimental setup and image optimization.
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Elson, L., Tjaden, A., Knapp, S., Müller, S. (2023). Characterization of Cellular Viability Using Label-Free Brightfield Live-Cell Imaging. In: Merk, D., Chaikuad, A. (eds) Chemogenomics. Methods in Molecular Biology, vol 2706. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3397-7_6
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DOI: https://doi.org/10.1007/978-1-0716-3397-7_6
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