Comet assay in reconstructed 3D human epidermal skin models--investigation of intra- and inter-laboratory reproducibility with coded chemicals
| Autor: | Andreas Zeller, Stefan Pfuhler, Gregory J. Carr, Raffaella Corvi, Thomas R. Downs, Kerstin Reisinger, Astrid A. Reus, Cyrille Krul |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Laboratory Proficiency Testing
Health Toxicology and Mutagenesis Drug Evaluation Preclinical Original Manuscript Phenylenediamines Toxicology medicine.disease_cause Models Biological Nitrophenols Tissue Culture Techniques In vivo Genetics medicine Humans Inter-laboratory Genetics (clinical) Carcinogen Reproducibility Chromatography integumentary system Chemistry Cyclohexanones Reproducibility of Results Methane sulfonate Methyl Methanesulfonate 4-Nitroquinoline-1-oxide Comet assay Ethylnitrosourea Comet Assay Epidermis Genotoxicity DNA Damage Mutagens |
| Zdroj: | Mutagenesis |
| ISSN: | 1464-3804 |
| Popis: | Reconstructed 3D human epidermal skin models are increasingly being used for safety testing of chemicals. Based on EpiDerm™ tissues, an assay was developed in which the tissues were topically exposed to test chemicals for 3h followed by cell isolation and assessment of DNA damage using the comet assay. Inter-laboratory reproducibility of the 3D skin comet assay was initially demonstrated using two model genotoxic carcinogens, methyl methane sulfonate (MMS) and 4-nitroquinoline-n-oxide (4NQO), and the results showed good concordance among three different laboratories and with in vivo data. In Phase 2 of the project, intra- and inter-laboratory reproducibility was investigated with five coded compounds tested at three different laboratories. For MMS and N-ethyl-N-nitrosourea (ENU), all laboratories found a dose-related and statistically significant increase (p30% cell loss), and the overall response was comparable in all laboratories despite some differences in doses tested. The results of the collaborative study for the coded compounds were generally reproducible among the laboratories involved and intra-laboratory reproducibility was also good. These data indicate that the comet assay in EpiDerm™ skin models is a relevant model for the safety assessment of compounds with a dermal route of exposure. JRC.I.5-Systems Toxicology |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|