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Nathalie Maubon, Bruno Loillier, Pascale Tuyaa-Boustugue, Annick Reboul and Philippe Masson. (Abbott Group, Laboratoires Fournier, Research Center, Global System Biology Department, HCS unit, 50 rue de Dijon, 21121 DAIX, France)

Development of a High Throughput Cytotoxicity Assay using High Content Screening Technology.

Despite massive investment in Pharmaceutical R&D, the number of new drugs obtaining approval has been declining in the past decade. While reasons behind such limited success are various, unwanted toxicity is one major reason for the failure rate. To address this problem, High Throughput Screening on cytotoxicity effect has to be added in the hit to lead phase during drug discovery. High-Content Screening (HCS) is the technique of choice to evaluate cytotoxic potential of compounds in a multiparametric environment. Recent papers have described a good correlation between in vivo and in vitro toxicity of compounds using HepG2 cells. In order to have a sufficiently high throughput, these assays were adapted, from 96w format to 384w format.
The assay development, using reference compounds activity on cell count, plasma membrane integrity and mitochondrial membrane potential were reported (Cytotox2 kit, Thermofisher). The assays were shown to be robust, giving Zā€™ of 0.75, 0.52 and 0.72 respectively; the whole process was set-up on a fully automated platform. Results of a screening run comprised of 3 000 compounds at 2 concentrations were obtained in one week.
In conclusion, a robust high-throughput cytotoxicity assay using multiparametric high content technology has been developed.
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