 | 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 platformindependent.
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.
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