The testis is a priority organ for developing alternative models to assess male reproductive health hazards of chemicals. This study characterized a 3D in vitro model of murine prepubertal Leydig TM3 cells with improved expression of steroidogenesis markers suitable for image-based screening of testicular toxicity. This 3D scaffold-free spheroid model was applied to explore the impact of prototypical endocrine-disrupting chemicals (EDCs) and environmental reprotoxicants (benzo[a]pyrene, 2- and 9-methylanthracenes, fluoranthene, triclosan, triclocarban, methoxychlor) on male reproductive health. The results were compared to the male reprotoxicity potential of EDCs assessed in a traditional monolayer (2D) culture. The testicular toxicity was dependent not only on the type of culture (2D vs. 3D models) but also on the duration of exposure. Benzo[a]pyrene and triclocarban were the most active compounds, eliciting cytotoxic effects in prepubertal Leydig cells at low micromolar concentrations, which might be a mechanism contributing to their male reprotoxicity.

睾丸是开发替代模型以评估化学品对男性生殖健康危害的优先器官。本研究对小鼠青春期前 Leydig TM3 细胞的 3D体外模型进行了表征，该模型具有改进的类固醇生成标记物的表达，适用于基于图像的睾丸毒性筛查。这种 3D 无支架球体模型用于探索原型内分泌干扰物 (EDC) 和环境生殖毒物（苯并[ a]芘、2-和9-甲基蒽、荧蒽、三氯生、三氯卡班、甲氧氯）对男性生殖健康的影响。将结果与在传统单层 (2D) 培养中评估的 EDC 的男性生殖毒性潜力进行了比较。睾丸毒性不仅取决于培养的类型（2D 与 3D 模型），还取决于暴露的持续时间。苯并[ a ]芘和三氯卡班是最活跃的化合物，它们在低微摩尔浓度下在青春期前间质细胞中引发细胞毒性作用，这可能是导致其男性生殖毒性的机制。