Validation of the 3D Skin Comet assay using full thickness skin models: Transferability and reproducibility

Publication date: March 2018 Source:Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Volume 827 Author(s): Kerstin Reisinger, Veronika Blatz, Joep Brinkmann, Thomas R. Downs, Anja Fischer, Frank Henkler, Sebastian Hoffmann, Cyrille Krul, Manfred Liebsch, Andreas Luch, Ralph Pirow, Astrid A. Reus, Markus Schulz, Stefan Pfuhler Recently revised OECD Testing Guidelines highlight the importance of considering the first site-of-contact when investigating the genotoxic hazard. Thus far, only in vivo approaches are available to address the dermal route of exposure. The 3D Skin Comet and Reconstructed Skin Micronucleus (RSMN) assays intend to close this gap in the in vitro genotoxicity toolbox by investigating DNA damage after topical application. This represents the most relevant route of exposure for a variety of compounds found in household products, cosmetics, and industrial chemicals. The comet assay methodology is able to detect both chromosomal damage and DNA lesions that may give rise to gene mutations, thereby complementing the RSMN which detects only chromosomal damage. Here, the comet assay was adapted to two reconstructed full thickness human skin models: the EpiDerm™- and Phenion® Full-Thickness Skin Models. First, tissue-specific protocols for the isolation of single cells and the general comet assay were transferred to European and US-American laboratories. After establishment of the assay, the protocol was then further optimized w...
Source: Mutation Research Genetic Toxicology and Environmental Mutagenesis - Category: Genetics & Stem Cells Source Type: research