A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy

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...

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Main Authors: Satoshi Yokota, Kiyoshi Hashimoto, Takuya Sato, Koichi Uemura, Kazuhide Makiyama, Takuya Nishimura, Satoshi Kitajima, Takehiko Ogawa
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:Current Research in Toxicology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666027X25000143
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author Satoshi Yokota
Kiyoshi Hashimoto
Takuya Sato
Koichi Uemura
Kazuhide Makiyama
Takuya Nishimura
Satoshi Kitajima
Takehiko Ogawa
author_facet Satoshi Yokota
Kiyoshi Hashimoto
Takuya Sato
Koichi Uemura
Kazuhide Makiyama
Takuya Nishimura
Satoshi Kitajima
Takehiko Ogawa
author_sort Satoshi Yokota
collection DOAJ
description 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.
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institution Kabale University
issn 2666-027X
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publishDate 2025-01-01
publisher Elsevier
record_format Article
series Current Research in Toxicology
spelling doaj-art-1122db72874e42ac9d9700eb4ead3bef2025-08-20T03:24:36ZengElsevierCurrent Research in Toxicology2666-027X2025-01-01810022810.1016/j.crtox.2025.100228A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapySatoshi Yokota0Kiyoshi Hashimoto1Takuya Sato2Koichi Uemura3Kazuhide Makiyama4Takuya Nishimura5Satoshi Kitajima6Takehiko Ogawa7Division of Cellular and Molecular Toxicology, Center for Biological Safety and Research, National Institute of Health Sciences, Kanagawa 210-9501, Japan; Corresponding authors at: Division of Cellular and Molecular Toxicology, Center for Biological Safety and Research, National Institute of Health Sciences, 3-25-26 Tono-machi, Kawasaki-ku, Kawasaki, Kanagawa 210-9501, Japan (Satoshi Yokota). Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa, 236-0004, Japan (Takehiko Ogawa).Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, Japan; Department of Urology, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, JapanDepartment of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, JapanDepartment of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, Japan; Department of Urology, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, JapanDepartment of Urology, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, JapanDivision of Cellular and Molecular Toxicology, Center for Biological Safety and Research, National Institute of Health Sciences, Kanagawa 210-9501, JapanDivision of Cellular and Molecular Toxicology, Center for Biological Safety and Research, National Institute of Health Sciences, Kanagawa 210-9501, JapanDepartment of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa 236-0004, Japan; Corresponding authors at: Division of Cellular and Molecular Toxicology, Center for Biological Safety and Research, National Institute of Health Sciences, 3-25-26 Tono-machi, Kawasaki-ku, Kawasaki, Kanagawa 210-9501, Japan (Satoshi Yokota). Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa, 236-0004, Japan (Takehiko Ogawa).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.http://www.sciencedirect.com/science/article/pii/S2666027X25000143Acrosin-GFP mouseTestis organ cultureTesticular toxicityRecoveryCisplatin
spellingShingle Satoshi Yokota
Kiyoshi Hashimoto
Takuya Sato
Koichi Uemura
Kazuhide Makiyama
Takuya Nishimura
Satoshi Kitajima
Takehiko Ogawa
A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
Current Research in Toxicology
Acrosin-GFP mouse
Testis organ culture
Testicular toxicity
Recovery
Cisplatin
title A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
title_full A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
title_fullStr A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
title_full_unstemmed A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
title_short A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
title_sort long term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
topic Acrosin-GFP mouse
Testis organ culture
Testicular toxicity
Recovery
Cisplatin
url http://www.sciencedirect.com/science/article/pii/S2666027X25000143
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