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P1015
CellCiphr – Cellular Systems Biology Assay for Hepatotoxicity Using Human HepG2 Cells
Presenter Stella Redpath, Millipore Corporation
Additional Authors: Andrew J. Ball, Janet L. Cuy, Kenneth A. Giuliano, William Irwin, Lawrence Vernetti, Patricia A. Johnston, Albert Gough, Dennis W. Harris, Rick Ryan, D. Lansing Taylor, Matthew Hsu
Drug-induced hepatotoxicity is a major factor in both the high fail rate of drug development and withdrawal of drugs from the market. Consequently, there is a need for more powerful test systems to identify potential hepatotoxins. Cellular Systems Biology (CSB)™ is the investigation of cells as integrated and interacting networks of genes, proteins and metabolic processes that give rise to either normal or pathological conditions. Cellular Systems Biology Assays utilize a panel of functional biomarkers to extract a cellular systemic response to a treatment or condition in the form of a panel of features measured in specific cell types, often with an High-content Screening (HCS) reader. Millipore and Cellumen have co-developed CellCiphr™ – a CSB assay for hepatotoxicity using human HepG2 cells. The assay utilizes HCS technology, and is platform-independent. The assay comprises detection reagents, protocols and software for profiling up to eleven hepatotoxicity endpoints, including Cell Loss, Cell Cycle Arrest, DNA Degradation/Apoptosis, Nuclear Size, Oxidative Stress, Stress Kinase Activation, DNA Damage, Mitochondrial Membrane Potential, Mitochondrial Mass, Mitotic Arrest and Cytoskeletal Integrity. The CellCiphr™ cytotoxicity kit allows analysis of all parameters at 3 time-points for 4 control toxins and up to 16 unknown compounds in two 10-point dose response curves. We present data showing that the reagents and protocols are suitable for analysis on widely-used HCS readers, such as the IN Cell Analyzer 1000 and ArrayScan™. Our data demonstrate the kit’s effectiveness in the detection of each parameter’s response to control toxins. The combination of unique antibody pairs and detection reagents allows comprehensive assessment and analysis of the cellular systemic response to hepatotoxin challenge. This CellCiphr™ assay filters compounds for toxicity and may be employed wherever compound ranking for efficacy or pre-clinical safety assessment is useful.