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THE USE OF 3D SKIN MODELS IN THE MICRONUCLEUS AND COMET ASSAYS: FURTHER PRE-VALIDATION STUDIES AND INVESTIGATIONS INTO INCREASING THROUGHPUT.

Pfuhler1, S., Ouedraogo2, G., Reisinger3, K., Krul4, C., Curren5, R., Aardema6, M., Fautz7, R., Barnett1, B., Downs1, T., Reuss4, A., Faquet2, B., Mun5, G., Hewitt8, N. 1The Procter & Gamble Co., Cincinnati, OH, USA; 2L’Oreal Life Sciences Research, Aulnay sous Bois, France; 3Henkel AG & Co KGaA, Duesseldorf, Germany; 4TNO, The Netherlands; 5Institute for In Vitro Sciences, Inc., Gaithersburg, MD, USA; 6Marilyn Aardema Consulting, LLC, USA; 7KPSS-Kao Professional Salon Services, Darmstadt, Germany; 8Erzhausen, Germany
Abstract

The European Cosmetic Association (COLIPA) has initiated a multi-laboratory project to establish and evaluate more predictive in vitro genotoxicity assays using 3D human tissues as replacements for current mammalian cell in vitro genotoxicity assays, which induce high levels of “false” positives. The reconstructed skin (RS) model, EpiDermTM, was combined with the micronucleus (MN) and Comet assays because the skin is the first site of contact of many different products, including cosmetics, thus the resulting RSMN model offers the potential for a more realistic application/metabolism of test compounds for evaluating genotoxicity [1,2]. Here, we show the progress and optimization of both assays as part of pre-validation studies.

Keywords

Comet assays, EpiDerm, Genotoxicity, Genotoxicity assays, Inter-laboratory reproducibility, Micronucleus assay, Multi-laboratory project, Pre-Validation, Reproducibility, RSMN assay, Transferability

Materials Tested

4-nitroquinoline N-oxide (4-NQO), 4-NQ, Cyclohexanone, Methyl methanesulfonate (MMS), Mitomycin C (MMC), MMS, N-ethyl-N-nitrosourea (ENU)

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