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Dr. Terry Riss, Promega Corporation, Madison

Cell-based Assays To Detect The Mechanism Of Toxicity

Abstract

Investigating cytotoxicity in vitro often begins with measuring of the number of viable cells remaining in cultures incubated with test compounds for 2-3 days. Alternatively, the number of dead cells can be measured by detecting markers that leak from dead cells into the culture medium. Currently available assay technologies can go beyond providing simple "live or dead" information to indicate whether cells have died by apoptosis or necrosis. More advanced assay methods can detect involvement of various organelles and cellular pathways that may trigger events leading to cell death. The knowledge of which pathways are involved provides important information that can lead to identification of the molecular targets involved with initiating cytotoxic events. This seminar will present an overview of methods of directly measuring cytotoxicity, detecting upstream pathways indirectly leading to cytotoxicity, and multiplexing assay combinations to more efficiently determine events leading to cell death and provide tips on how to choose the best assay.

Dr Terry Riss started the Cell Biology program at Promega Corporation in 1990. He has since held several R&D and Project Management positions. Dr. Riss managed development of cell viability, cytotoxicity, apoptosis, and protease assay systems and also lead efforts to identify and promote multiplexing of cell-based assays to determine the mechanism of cell death. Dr. Riss now serves as Senior Product Specialist, Cell Health involved in outreach educational training activities. Dr. Riss regularly participates in NIH study sections reviewing HTS grants and is co-editor of the cell culture assays section of the Assay Guidance Manual hosted by the NIH.

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