Toxicity assays in nanodrops combining bioassay and morphometric endpoints
| Autor: | Jesús Angulo, Frédéric Lemaire, Julien Reboud, Christian Lajaunie, André-Patrick Arrigo, Chantal Diaz-Latoud, Béatrice Schaack, Hervé Pointu, Alexandre Papine, Céline A. Mandon, Francois Chatelain |
|---|---|
| Přispěvatelé: | DRDC/BIM [Grenoble], CEA-Grenoble, Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, IMSTAR, IMSTAR S.A., Centre de Morphologie Mathématique (CMM), MINES ParisTech - École nationale supérieure des mines de Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Centre de Bioinformatique (CBIO), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Mines Paris - PSL (École nationale supérieure des mines de Paris) |
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
MESH: Cell Death
lcsh:Medicine 010501 environmental sciences MESH: Heat-Shock Proteins 01 natural sciences MESH: Dose-Response Relationship Drug MESH: Hepatocytes MESH: Paraquat Cadmium Chloride Bioassay MESH: Animals Promoter Regions Genetic lcsh:Science Heat-Shock Proteins MESH: Cadmium Chloride media_common 0303 health sciences Multidisciplinary Cell Death Heavy metals Limiting MESH: Gene Expression Regulation MESH: Environmental Pollutants Toxicity Arsenates Biological Assay Environmental Pollutants MESH: Arsenates Research Article Biotechnology Drug Paraquat MESH: High-Throughput Screening Assays media_common.quotation_subject Green Fluorescent Proteins Computational biology [SDV.BC]Life Sciences [q-bio]/Cellular Biology Biology MESH: Biological Assay Cell Line 03 medical and health sciences MESH: Nanostructures MESH: Green Fluorescent Proteins High-Throughput Screening Assays Toxicity Tests MESH: Promoter Regions Genetic Animals Humans MESH: Toxicity Tests 030304 developmental biology 0105 earth and related environmental sciences MESH: Herbicides MESH: Humans Dose-Response Relationship Drug business.industry Herbicides lcsh:R Cell Biology Microarray Analysis Nanostructures MESH: Cell Line MESH: Microarray Analysis Gene Expression Regulation Hepatocytes lcsh:Q business |
| Zdroj: | PLoS ONE PLoS ONE, Public Library of Science, 2007, 2 (1), pp.e163. ⟨10.1371/journal.pone.0000163⟩ PLoS ONE, 2007, 2 (1), pp.e163. ⟨10.1371/journal.pone.0000163⟩ PLoS ONE, Vol 2, Iss 1, p e163 (2007) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0000163⟩ |
| Popis: | BACKGROUND: Improved chemical hazard management such as REACH policy objective as well as drug ADMETOX prediction, while limiting the extent of animal testing, requires the development of increasingly high throughput as well as highly pertinent in vitro toxicity assays. METHODOLOGY: This report describes a new in vitro method for toxicity testing, combining cell-based assays in nanodrop Cell-on-Chip format with the use of a genetically engineered stress sensitive hepatic cell line. We tested the behavior of a stress inducible fluorescent HepG2 model in which Heat Shock Protein promoters controlled Enhanced-Green Fluorescent Protein expression upon exposure to Cadmium Chloride (CdCl2), Sodium Arsenate (NaAsO2) and Paraquat. In agreement with previous studies based on a micro-well format, we could observe a chemical-specific response, identified through differences in dynamics and amplitude. We especially determined IC50 values for CdCl2 and NaAsO2, in agreement with published data. Individual cell identification via image-based screening allowed us to perform multiparametric analyses. CONCLUSIONS: Using pre/sub lethal cell stress instead of cell mortality, we highlighted the high significance and the superior sensitivity of both stress promoter activation reporting and cell morphology parameters in measuring the cell response to a toxicant. These results demonstrate the first generation of high-throughput and high-content assays, capable of assessing chemical hazards in vitro within the REACH policy framework. |
| Databáze: | OpenAIRE |
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