Evaluation of the transferability of in vivo Pig-a gene mutation assay across three laboratories (생체 내 Pig-a 유전자 돌병변이시험에 대한 세 시험실의 전수가능성 평가)

Young-Shin Chung (정영신); Bum-Soo Park (박범수); Se-Hee Han (한세희); Ji-Yeon Lee (이지연); Ji-Young Kim (김지영); Seung-Min Back (백승민); Cho-Rong Park (박초롱); Su-Hwan Kim (김수환); Jong-Kwon Lee (이종권)

doi:10.23032/jaae.2017.11.1.004

Evaluation of the transferability of in vivo Pig-a gene mutation assay across three laboratories

Journal of Alternatives to Animal Experiments
Abbr : J Altern Anim Exp
2017, 11(1), pp.1-8
DOI : 10.23032/jaae.2017.11.1.004
Publisher : Korean Society for Alternatives to Animal Experiments
Research Area : Medicine and Pharmacy > Veterinary > Basic Veterinary Medicine > Laboratory Animal Medicine
Received : November 28, 2017
Accepted : December 21, 2017
Published : December 29, 2017

Young-Shin Chung ¹, Bum-Soo Park ¹, Se-Hee Han ¹, Ji-Yeon Lee ¹, Ji-Young Kim ², Seung-Min Back ², Cho-Rong Park ³, Su-Hwan Kim ³, Jong-Kwon Lee ⁴

¹호서대학교 생명공학과
²안전성평가연구소
³(주)바이오톡스텍
⁴식품의약품안전평가원 특수독성과

ABSTRACT

In order to establish a pig-a gene mutation assay as a regulatory test for evaluating in vivo genotoxicity in Korea, the transferability study as a stage II validation was performed using the test substance, N-ethyl-N-nitrosourea (ENU) across three laboratories. The lead laboratory (KIT) standardized the protocol using Rat MutaFlow® Kit and transferred the methodology to two participating laboratories (Hoseo University and Biotoxtech Co. Ltd.). Sprague Dawley rats were treated with vehicle or three dose levels of ENU (10, 20, 40mg/kg) by oral gavages on the 3 consecutive days. On the day 15 after the first treatment, bloods were collected and analyzed to estimate the percentage of reticulocytes (%RETs), mutant type of RETs and erythrocytes (RBC) using flowcytometer. In results, the exposure of ENU induced the dose-dependent increases in the mutant frequencies of RBC CD59- and RET CD59- , while %RETs were not affected. Data comparisons among three participating laboratories indicated the good transferability with the high correlation coefficients (0.97-0.99) between the lead and participating laboratories. In conclusion, the pig-a gene mutation assay may be the effective test for the estimation of in vivo gene mutation and the validation studies may be needed using more test substances in Korea.

KEYWORDS

Pig-a gene, CD59, in vivo mutagenecity, N-ethyl-N-nitrosourea, transferability

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Journal of Alternatives to Animal Experiments 2023 KCI Impact Factor : 0.07

Evaluation of the transferability of in vivo Pig-a gene mutation assay across three laboratories

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* References for papers published after 2023 are currently being built.

Journal of Alternatives to Animal Experiments 2023 KCI Impact Factor : 0.07

Evaluation of the transferability of in vivo Pig-a gene mutation assay across three laboratories

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