Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells

摘要: Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they still an early stage. Consequently, we aimed develop male animals using mouse spermatogonial stem cells (mSSCs). Here, modified OECD TG 489 optimized vitro comet assay our previous study. This study verify validity of tests involving mSSCs by comparing their results with those vivo C57BL/6 mice gavage. We selected hydroxyurea (HU), which is known chemically induce toxicity. The 50% inhibitory concentration (IC50) value HU was 0.9 mM, as determined MTT assay. In assay, % tail DNA Olive moment (OTM) after administration increased significantly, compared control. Annexin V, PI staining TUNEL assays showed that caused apoptosis mSSCs. order compare tests, same substances were administered mice. Reproductive toxicity observed at 25, 50, 100, 200 mg/kg/day measured clinical measures reduction sperm motility testicular weight. DCFH-DA H&E staining, also performed. similar for treatment. Finally, linear regression analysis a strong positive correlation between vivo. conclusion, present first demonstrate effect HU-induced damage, ROS formation, Further, current suggest could be useful model predict