Poster – Use of Automated High Content Analysis or Imaging Applied To Assessment Of Primary DNA Damage With γH2AX and Comet Assays on different Cell types

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Genotoxicity potential assessment of pharmaceuticals is mandatory for registration. In addition to the standard in vitro/ in vivo test battery, other assays can be of interest because of their high throughput in the drug discovery stage but also for mechanistic understanding.

Different other assays can be performed to detect DNA damage, such as γH2AX and the single cell gel electrophoresis (or comet assay).

The γH2AX assay was suggested as a new in vitro assay for detecting genotoxic properties of chemicals. The phosphorylation of γH2AX has been demonstrated to be a sensitive marker for a wide range of DNA damage. In this study, primary human keratinocytes were evaluated to provide a more relevant testing system for genotoxicity assessment in the skin for topical dermatological/cosmetic substances. Primary human keratinocytes were compared to the well known HepG2 cell line for which data have already been published.

The comet assay is a sensitive, well established technique for quantifying DNA damage in eukaryotic cells and enabling the detection of a wide range of DNA damaging agents. The assay can be adapted in a 96 wells format allowing higher throughput.

The high content imaging (HCI) analysis applied to genotoxicity assays is a valuable tool allowing higher throughput and multi-parametric analysis in order to improve predictability of these assays.

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