title: Reference control data obtained from an in vivo comet-micronucleus combination assay using Sprague Dawley rats.
abstract: According to the International Conference on Harmonization Guidance on Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use (ICH S2(R1)), a positive response in any in vitro assay necessitates additional in vivo test(s) (other tissue/endpoint) in addition to the erythrocyte micronucleus test when Option 1 of the test battery is selected. When Option 2 of the test battery is selected, a bacterial gene mutation test and two in vivo tests with different tissues/endpoint are required. The in vivo alkaline comet assay is recommended as the second in vivo test because it can detect a broad spectrum of DNA damage in any tissue and can be combined with the erythrocyte micronucleus test. Considering animal welfare, a combination assay is preferable to an individual assay. Thus, we validated the protocol for the in vivo comet-micronucleus combination assay in rats with three daily administrations and determined the dose of the positive control (ethyl methanesulfonate; EMS, 200mg/kg/day). We also collected the negative control (vehicle) and positive control (EMS) data from the comet (liver, stomach, and kidney) and micronucleus (bone marrow) combination assay using male Sprague Dawley (SD) rats. The negative control data were comparable to our historical control data obtained from stand-alone assays. The positive control data showed clear and consistent positive responses in both endpoints.
authors.affiliation: Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, Shioshinden, Iwata, Shizuoka, 437-1213, Japan. Electronic address: kasamoto-sawako@anpyo.or.jp.
authors.forename: Sawako
authors.initials: S
authors.lastname: Kasamoto
authors.affiliation: Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, Shioshinden, Iwata, Shizuoka, 437-1213, Japan.
authors.forename: Shoji
authors.initials: S
authors.lastname: Masumori
authors.affiliation: Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, Shioshinden, Iwata, Shizuoka, 437-1213, Japan.
authors.forename: Jin
authors.initials: J
authors.lastname: Tanaka
authors.affiliation: Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, Shioshinden, Iwata, Shizuoka, 437-1213, Japan.
authors.forename: Maya
authors.initials: M
authors.lastname: Ueda
authors.affiliation: Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, Shioshinden, Iwata, Shizuoka, 437-1213, Japan.
authors.forename: Masahito
authors.initials: M
authors.lastname: Fukumuro
authors.affiliation: Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, Shioshinden, Iwata, Shizuoka, 437-1213, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Nagai
authors.affiliation: Osaka Prefecture University, 1-58, Rinku Ourai Kita, Izumisano, Osaka, 598-8531, Japan.
authors.forename: Jyoji
authors.initials: J
authors.lastname: Yamate
authors.affiliation: Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, Shioshinden, Iwata, Shizuoka, 437-1213, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Hayashi
mesh_terms: D000818:Animals; D020552:Comet Assay / Q000379:methods*; D035061:Control Groups; D005020:Ethyl Methanesulfonate / Q000633:toxicity; D008297:Male; D015162:Micronucleus Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D009153:Mutagens; D005020:Ethyl Methanesulfonate
keywords: Combination assay; Comet assay; Historical control data; Micronucleus test; Protocol