연구논문 : 수컷 생식줄기세포를 이용한 생식독성 동물대체시험법 개발
= Original Articles : Development of an Alternative Test Method for the Evaluation of Reproductive Toxicity Using Male Germline Stem Cells

 
제어번호 101795134
저자명 전혜련 ( Hye Lyun Jeon )  , 김태성 ( Tae Sung Kim )  , 이정선 ( Jung Sun Yi )  , 안일영 ( Il Young Ahn )  ,

           고경육 ( Kyung Yuk Ko )  , 이진하 ( Jin Ha Lee )  , 김주환 ( Joo Hwan Kim )  , 손수정 ( Soo Jung Sohn ) 
학술지명 동물실험대체법학회지
권호사항 Vol.9 No.1 [2015] 
발행처 한국동물실험대체법학회(N/A) 
자료유형 학술저널
수록면 5-22(18쪽)
언어 Korean
발행년도 2015년
판매처 한국학술정보

 
초록
Currently, alternative test methods are actively being developed as a replacement for animal testing, based on the 3Rs (Replacement, Refinement, Reduction). However, the development of alternative test methods for the evaluation of reproductive and developmental toxicity is in its early stage, and no established test methods exist. This study is aimed at developing an alternative test method to evaluate reproductive toxicity using male germline stem cells (GSC). We selected a negative toxic substance (Dimethyl sufloxide (DMSO)) and two positive toxic substances (NEthyl- N-Nitrosourea (ENU), methyl methanesulfonate (MMS) characterized in OECD TG 489. We also used seven test substances (2,4-diaminotoluene (2,4-DAT), cyclophosphamide (CP), benzo[α]pyrene (BP), cadmium chloride (CdCl2), D-mannitol (MA), n-butyl chloride (NBC) and trimethyl ammonium chloride (TAC)) suggested in a scientific paper published by ECVAM. The endpoints of toxicological evaluation were cell viability (MTT assay) and comet assay which is a method to measure DNA damage. As a result of our study with a 50% inhibitory concentration (IC50) determined using the MTT assay, IC50 values of ENU and MMS were 1.7 mM and 0.4 mM, respectively. Also, IC50 values of 2,4-DAT, CP, BP andCdCl2 were 10.3 mM, 5.5 mM, 0.4 mM and 0.18 mM, respectively. As cell viability wasn’t significantly different from that of the control, IC50 values of MA, NBC and TAC could not be calculated. In the comet assay, Tail DNA%, Tail Length (TL) and Olive Tail Moment (OTM) of the two positive toxic substances (ENU and MMS) and the four test substances (2,4-DAT, CP, BP and CdCl2) significantly grew in comparison with the control. However, Tail DNA%, TL and OTM of the negative toxic substance (DMSO) and the three positive toxic substances (MA, NBC, TAC) were similar to those of the control. In conclusion, this study demonstrated that the comet assay using GSC could be a candidate test method in predicting male reproductive toxicity.

주제어
Alternative test method  , Male germline stem cell  , Reproductive toxicity  , Comet assay  , DNA damage