Yokota Satoshi, Hashimoto Kiyoshi, Sato Takuya, Uemura Koichi, Makiyama Kazuhide, Nishimura Takuya, Kitajima Satoshi, Ogawa Takehiko
Division of Cellular and Molecular Toxicology, Center for Biological Safety and Research, National Institute of Health Sciences, Kanagawa 210-9501, Japan.
Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, Japan.
Curr Res Toxicol. 2025 Mar 11;8:100228. doi: 10.1016/j.crtox.2025.100228. eCollection 2025.
We previously developed the acrosin-green fluorescent protein (GFP) transgenic neonatal mouse organ culture system for rapid and accurate assessment of testicular toxicity. This system effectively evaluates drug-induced toxicity in male germ cells before meiotic entry but cannot assess post-meiotic germ cell toxicity. For many chemicals, the specific stage of germ cell differentiation that is susceptible to toxicity remains unclear, highlighting the need for new methods. In this study, we incubated neonatal mouse testis organ cultures for 35 days to allow post-meiotic cells to develop. The tissue was then exposed to cisplatin to determine the cells that are targeted and to assess the reversibility of the toxicity. We monitored changes in tissue volume and GFP fluorescence, which tracks the progression of spermatogenesis, and confirmed findings by histological analysis. Cisplatin inhibited tissue growth and reduced GFP fluorescence in a concentration-dependent manner. Higher concentrations targeted not only spermatogonia, but also spermatocytes and spermatids. Recovery from toxicity was observed at clinically relevant doses. This study demonstrates that long-term mouse testis organ culture can be used to assess testicular toxicity, enabling the identification of specific germ cell stages targeted by chemicals such as cisplatin.
我们之前开发了顶体蛋白酶-绿色荧光蛋白（GFP）转基因新生小鼠器官培养系统，用于快速准确地评估睾丸毒性。该系统能有效评估减数分裂开始前雄性生殖细胞中的药物诱导毒性，但无法评估减数分裂后生殖细胞的毒性。对于许多化学物质而言，易受毒性影响的生殖细胞分化的具体阶段仍不清楚，这凸显了对新方法的需求。在本研究中，我们将新生小鼠睾丸器官培养35天，以使减数分裂后的细胞发育。然后将组织暴露于顺铂，以确定被靶向的细胞，并评估毒性的可逆性。我们监测了组织体积和跟踪精子发生进程的GFP荧光的变化，并通过组织学分析证实了研究结果。顺铂以浓度依赖的方式抑制组织生长并降低GFP荧光。较高浓度不仅靶向精原细胞，还靶向精母细胞和精子细胞。在临床相关剂量下观察到了毒性恢复。本研究表明，长期小鼠睾丸器官培养可用于评估睾丸毒性，从而能够识别顺铂等化学物质所靶向的特定生殖细胞阶段。
