지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2017.4
- 수록면
- 107 - 118 (12page)
이용수
초록· 키워드
오류제보하기
Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration (IC50) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.
목차
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
참고문헌 (38)
참고문헌 신청
Rovida, C / 2009 / Re-evaluation of animal numbers and costs for in vivo tests to accomplish REACH legislation requirements for chemicals - a report by the transatlantic think tank for toxicology (t(4)) / ALTEX 26:187~208
Schulte, R.T / 2010 / Sperm DNA damage in male infertility: etiologies, assays, and outcomes / J. Assist. Reprod. Genet 27:3~12
Makker, K / 2009 / Oxidative stress & male infertility / Indian J. Med. Res 129:357~367
Adler, S / 2010 / Alternative (non-animal)methods for cosmetics testing: current status and future prospects-2010 / Arch. Toxicol 85:367~485
Tegelenbosch, R.A / 1993 / A quantitative study of spermatogonial multiplication and stem cell renewal in the C3H/101 F1 hybrid mouse / Mutat. Res 290:193~200
Schulte, R.T / 2010 / Sperm DNA damage in male infertility: etiologies, assays, and outcomes / J. Assist. Reprod. Genet 27:3~12
Makker, K / 2009 / Oxidative stress & male infertility / Indian J. Med. Res 129:357~367
Adler, S / 2010 / Alternative (non-animal)methods for cosmetics testing: current status and future prospects-2010 / Arch. Toxicol 85:367~485
Tegelenbosch, R.A / 1993 / A quantitative study of spermatogonial multiplication and stem cell renewal in the C3H/101 F1 hybrid mouse / Mutat. Res 290:193~200
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UCI(KEPA) : I410-ECN-0101-2017-513-002410025