title: High-content screening imaging and real-time cellular impedance monitoring for the assessment of chemical's bio-activation with regards hepatotoxicity.
abstract: Testing hepatotoxicity is a crucial step in the development and toxicological assessment of drugs and chemicals. Bio-activation can lead to the formation of metabolites which may present toxicity for the organism. Classical cytotoxic tests are not always appropriate and are often insufficient, particularly when non metabolically-competent cells are used as the model system, leading to false-positive or false-negative results. We tested over 24 h the effects of eight reference compounds on two different cell models: primary cultures of rat hepatocytes and FAO hepatoma cells that lack metabolic properties. We performed inter-assay validation between three classical cytotoxicity assays and real-time cell impedance data. We then complemented these experiments with high-content screening (HCS) to determine the cell function disorders responsible for the observed effects. Among the different assays used, the neutral red test seemed to be well suited to our two cell models, coupled with real-time cellular impedance which proved useful in the detection of bio-activation. Indeed, impedance monitoring showed a high sensitivity with interesting curve profiles yet seemed unsuitable for evaluation of viability on primary culture. Finally, HCS in the evaluation of hepatotoxicity is likely to become an essential tool for use in parallel to a classical cytotoxic assay in the assessment of drugs and environmental chemicals.
authors.affiliation: UMR 1331 TOXALIM (Research Centre in Food Toxicology), Institut National de la Recherche Agronomique (INRA), Laboratory of Xenobiotic's Cellular and Molecular Toxicology, 400 route des Chappes, BP 167, 06903 Sophia-Antipolis Cedex, France. Electronic address: ludovic.peyre@sophia.inra.fr.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Peyre
authors.affiliation: UMR 1331 TOXALIM (Research Centre in Food Toxicology), Institut National de la Recherche Agronomique (INRA), Laboratory of Xenobiotic's Cellular and Molecular Toxicology, 400 route des Chappes, BP 167, 06903 Sophia-Antipolis Cedex, France.
authors.forename: Georges
authors.initials: G
authors.lastname: de Sousa
authors.affiliation: Odesia NeoSciences, INRA-Agrobiotech, 06903 Sophia-Antipolis, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Barcellini-Couget
authors.affiliation: Galderma R&D, 2400 Route des Colles, BP87, 06902 Sophia Antipolis Cedex, France.
authors.forename: Anne-Pascale
authors.initials: AP
authors.lastname: Luzy
authors.affiliation: UMR 1331 TOXALIM (Research Centre in Food Toxicology), Institut National de la Recherche Agronomique (INRA), Laboratory of Xenobiotic's Cellular and Molecular Toxicology, 400 route des Chappes, BP 167, 06903 Sophia-Antipolis Cedex, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Zucchini-Pascal
authors.affiliation: UMR 1331 TOXALIM (Research Centre in Food Toxicology), Institut National de la Recherche Agronomique (INRA), Laboratory of Xenobiotic's Cellular and Molecular Toxicology, 400 route des Chappes, BP 167, 06903 Sophia-Antipolis Cedex, France. Electronic address: Roger.rahmani@sophia.inra.fr.
authors.forename: Roger
authors.initials: R
authors.lastname: Rahmani
mesh_terms: D000082:Acetaminophen / Q000633:toxicity; D000655:Amodiaquine / Q000633:toxicity; D000818:Animals; D002220:Carbamazepine / Q000633:toxicity; D045744:Cell Line, Tumor; D002470:Cell Survival; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury; D004054:Diethylstilbestrol / Q000633:toxicity; D004917:Erythromycin / Q000633:toxicity; D005665:Furosemide / Q000633:toxicity; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D057166:High-Throughput Screening Assays; D008297:Male; D009499:Neutral Red / Q000378:metabolism; D051381:Rats; D018675:Toxicity Tests; D014212:Tretinoin / Q000633:toxicity
chemical_list: D000655:Amodiaquine; D009499:Neutral Red; D002220:Carbamazepine; D000082:Acetaminophen; D014212:Tretinoin; D004917:Erythromycin; D004054:Diethylstilbestrol; D005665:Furosemide
keywords: Bio-activation; Cytotoxic assays; Drug-induced liver injury; Hepatotoxicity; High-content screening; Real-time cellular impedance