Intra- and inter-laboratory reproducibility and accuracy of the LuSens assay: A reporter gene-cell line to detect keratinocyte activation by skin sensitizers.

Autor: Ramirez T; BASF SE, Experimental Toxicology and Ecology, Germany., Stein N; BASF SE, Experimental Toxicology and Ecology, Germany., Aumann A; BASF SE, Experimental Toxicology and Ecology, Germany., Remus T; DSM Nutritional Products AG, Kaiseraugst, Switzerland., Edwards A; Burleson Research Technologies, Morrisville, NC, USA., Norman KG; Institute for In Vitro Sciences, Gaithersburg, MD, USA., Ryan C; The Procter and Gamble Company, Mason, OH, USA., Bader JE; Institute for In Vitro Sciences, Gaithersburg, MD, USA., Fehr M; DSM Nutritional Products AG, Kaiseraugst, Switzerland., Burleson F; Burleson Research Technologies, Morrisville, NC, USA., Foertsch L; The Procter and Gamble Company, Mason, OH, USA., Wang X; The Procter and Gamble Company, Mason, OH, USA., Gerberick F; The Procter and Gamble Company, Mason, OH, USA., Beilstein P; DSM Nutritional Products AG, Kaiseraugst, Switzerland., Hoffmann S; seh consultancy + services, Paderborn, Germany., Mehling A; BASF Personal Care and Nutrition GmbH, Germany., van Ravenzwaay B; BASF SE, Experimental Toxicology and Ecology, Germany., Landsiedel R; BASF SE, Experimental Toxicology and Ecology, Germany. Electronic address: robert.landsiedel@basf.com.
Jazyk: angličtina
Zdroj: Toxicology in vitro : an international journal published in association with BIBRA [Toxicol In Vitro] 2016 Apr; Vol. 32, pp. 278-86. Date of Electronic Publication: 2016 Jan 12.
DOI: 10.1016/j.tiv.2016.01.004
Abstrakt: Several non-animal methods are now available to address the key events leading to skin sensitization as defined by the adverse outcome pathway. The KeratinoSens assay addresses the cellular event of keratinocyte activation and is a method accepted under OECD TG 442D. In this study, the results of an inter-laboratory evaluation of the "me-too" LuSens assay, a bioassay that uses a human keratinocyte cell line harboring a reporter gene construct composed of the rat antioxidant response element (ARE) of the NADPH:quinone oxidoreductase 1 gene and the luciferase gene, are described. Earlier in-house validation with 74 substances showed an accuracy of 82% in comparison to human data. When used in a battery of non-animal methods, even higher predictivity is achieved. To meet European validation criteria, a multicenter study was conducted in 5 laboratories. The study was divided into two phases, to assess 1) transferability of the method, and 2) reproducibility and accuracy. Phase I was performed by testing 8 non-coded test substances; the results showed a good transferability to naïve laboratories even without on-site training. Phase II was performed with 20 coded test substances (performance standards recommended by OECD, 2015). In this phase, the intra- and inter-laboratory reproducibility as well as accuracy of the method was evaluated. The data demonstrate a remarkable reproducibility of 100% and an accuracy of over 80% in identifying skin sensitizers, indicating a good concordance with in vivo data. These results demonstrate good transferability, reliability and accuracy of the method thereby achieving the standards necessary for use in a regulatory setting to detect skin sensitizers.
(Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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