 | 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. High Content Screening enables the
investigation of cells as integrated and interacting
networks of genes, proteins and metabolic processes
that give rise to either normal or pathological conditions.
Though multiplexing of high specificity detection
reagents, High Content Screening assays can extract a
systemic cellular response to a treatment or condition in
the form of a panel of features measured in specific cell
types. Millipore has developed a High Content Screening
assay for hepatotoxicity using human HepG2 cells. The
assay comprises detection reagents and protocols 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 commercially available assay 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 GE IN Cell
Analyzer 1000 and ThermoFisher 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 response to hepatotoxin challenge. The
assay may be used to filter compounds for toxicity and
thus has utility for compound ranking for efficacy or preclinical
safety assessment.
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