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).

keratinocyte2The 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.

View and download on Slideshare (low quality) : http://fr.slideshare.net/hcspharma/use-of-automated-high-content-analysis-or-imaging-applied-to-assessment-of-primary-dna-damage-with-h2ax-and-comet-assays-on-different-cell-types

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Cell biology is part of the R&D activity in dermocosmetology. At HCS Pharma, we are developing new democosmetology assays on human primary keratinocytes using our automation plateform and high content analysis suite.

Assessment of compounds activity/safety can be performed on several parameters (wound healing, hydration, inflammation, toxicity, genotoxicity). Further parameters could be analyzed on demand using probes and immunocytochemistry.

Thought the use of high content analysis technologies, we can provide a wide range of results from phenotypes quantification cell by cell to live cells imaging movies.

View and download on Slideshare (low quality) : http://fr.slideshare.net/hcspharma/in-vitro-dermocosmetology-high-content-analysis-approach-using-human-primary-keratinocytes

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If you are in Cosm’ing meeting, don’t hesitate to go and discuss with Julian BURSZTYKA, our COO. He presents our work in HCS Pharma by showing a poster on genotoxicity assay, developped in collaboration with Toxicology in vitro team of Galderma R&D and he will give a presentation at 9.15 am on the interest of using cell imaging for your R&D projects.

Bannière cosm'ing

HCSPharma-Galderma

During ELRIGfr event in Rennes, We had the opportunity to present our work on genotoxicity assay (comet assay on TK6 & gH2AX on keratinocytes and HepG2) in collaboration with the team of in vitro of Galderma R&D. It is really exciting to work with them since they are really experts in their domain and they want to work on innovative solutions in HCS. We really want to thank them for our interesting discussion and our collaborative projects!

HCSPharma-Galderma

PosterGenotoxHCSPharma-Galderma

Quantifying DNA damage is mandatory to assess potential adverse effects of candidate drugs or molecules or extracts developed in the dermo-cosmetic industry, but also to assess the efficacy of therapeutic approaches with the aim of producing tumor cell genotoxicity in cancer treatment.

The comet assay is a sensitive, well established technique for quantifying DNA damage in eukaryotic cells. Compatible with the detection of a wide range of DNA damaging agents, its principle consists in the migration of fragmented DNA in an electrophoresis gel (damaged DNA forming the tail of the comet), while intact DNA moves at a slower rate (head of the comet). The percentage of fragmented DNA in the comet tail is a direct measure of DNA damage.

View and download on Slideshare (low quality) :
http://fr.slideshare.net/hcspharma/development-of-an-automated-comet-assay-for-genotoxicity-assessment-on-tk6-cell-line

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I was present at the last congress ELRIGfr in Brussels. It was a strong exchange place with interesting presentations around robotics. It is always really interesting to see examples of innovative automation which give really robust results. For example, Frank Gudermann from university of Bielefield in Germany showed that automation of culture cell with cell count by holographic picture gives more robust results as manual count by trypan blue. Or automation of PK sampling with nanoliter by using echo technology allows increasing throughput of bioanalytical analysis for PK studies in Hoffmann La Roche. Also, another presentation of IGR has shown a complete robotic platform with 4 imagers (Micro from MDS) and 1 FACS totally automated from cell seeding to staining and reading.

It was a great pleasure also for us to present 4 different posters on our few last month works in cell imaging. This work was done in collaboration with Perkin Elmer on the Operetta machine. This machine allows us to take images in a 3D culture as shown in these posters: neuroprotection model for parkinson disease and oncology and performed also really nice images showed in these other posters: hepatotoxicity assay and genotoxicity assays.

Nathalie and Julian were present during ELRIGfr event last week. They presented their company, their work and their collaborations. They have also presented two posters : the first one on a co-culture model of Parkinson and first results in 3 dimensions ; the other one on genotoxicity and steatosis assays on HepG2 cells. These next two to three months will be dedicated to validate these assays. New assays  will also be undertaken, like transporters assays, cholestasis or phospholipidosis, on HepG2 cells as well as on primary hepatotocytes. Genotoxicity assays are under validation on different cell lines as HepG2, SH-SY5Y, HeLa, HT29 cells. If you want more details, don’t hesitate to contact us.

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