Germ cell depletion using HSV-TK in mouse testes

Abstract Germ cell transplantation is useful for the study of male germ cells and the generation of genetically modified animals. For transplantation, germ cell-free hosts generated using anticancer drug treatment, irradiation exposure, or genetic mutation are required. In this study, we aimed to de...

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Main Authors: Constance Dollet, Miyuki Shindo, Shun Takahashi, Kento Ito, Tomoo Eto, Toshiaki Watanabe
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-93325-6
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author Constance Dollet
Miyuki Shindo
Shun Takahashi
Kento Ito
Tomoo Eto
Toshiaki Watanabe
author_facet Constance Dollet
Miyuki Shindo
Shun Takahashi
Kento Ito
Tomoo Eto
Toshiaki Watanabe
author_sort Constance Dollet
collection DOAJ
description Abstract Germ cell transplantation is useful for the study of male germ cells and the generation of genetically modified animals. For transplantation, germ cell-free hosts generated using anticancer drug treatment, irradiation exposure, or genetic mutation are required. In this study, we aimed to develop a new system for germ cell depletion, more in compliance with the “3R” principles. For this purpose, we generated knock-in mice expressing a subtype of the herpes simplex virus type 1 thymidine kinase (HSV-TK30), reported to not induce infertility, unlike the original HSV-TK gene. Ganciclovir injection resulted in nearly complete abrogation of spermatogenesis. Furthermore, transplanted spermatogonial stem cells were differentiated into sperm in the host testes, and they gave rise to offspring. Therefore, the mice developed in this study enable the efficient removal of germ cells for germ cell transplantation in a manner more compliant with the 3R principles.
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spelling doaj-art-2b912a4fed364685ae9e3c8fddcd83ee2025-08-20T02:41:31ZengNature PortfolioScientific Reports2045-23222025-03-0115111110.1038/s41598-025-93325-6Germ cell depletion using HSV-TK in mouse testesConstance Dollet0Miyuki Shindo1Shun Takahashi2Kento Ito3Tomoo Eto4Toshiaki Watanabe5Center for Regenerative Medicine, National Center for Child Health and DevelopmentCenter for Regenerative Medicine, National Center for Child Health and DevelopmentCenter for Regenerative Medicine, National Center for Child Health and DevelopmentCenter for Regenerative Medicine, National Center for Child Health and DevelopmentCentral Institute for Experimental Medicine and Life ScienceCenter for Regenerative Medicine, National Center for Child Health and DevelopmentAbstract Germ cell transplantation is useful for the study of male germ cells and the generation of genetically modified animals. For transplantation, germ cell-free hosts generated using anticancer drug treatment, irradiation exposure, or genetic mutation are required. In this study, we aimed to develop a new system for germ cell depletion, more in compliance with the “3R” principles. For this purpose, we generated knock-in mice expressing a subtype of the herpes simplex virus type 1 thymidine kinase (HSV-TK30), reported to not induce infertility, unlike the original HSV-TK gene. Ganciclovir injection resulted in nearly complete abrogation of spermatogenesis. Furthermore, transplanted spermatogonial stem cells were differentiated into sperm in the host testes, and they gave rise to offspring. Therefore, the mice developed in this study enable the efficient removal of germ cells for germ cell transplantation in a manner more compliant with the 3R principles.https://doi.org/10.1038/s41598-025-93325-6Germ cell transplantationSpermatogonial stem cellsHost miceTestesHSV-TKDDX4
spellingShingle Constance Dollet
Miyuki Shindo
Shun Takahashi
Kento Ito
Tomoo Eto
Toshiaki Watanabe
Germ cell depletion using HSV-TK in mouse testes
Scientific Reports
Germ cell transplantation
Spermatogonial stem cells
Host mice
Testes
HSV-TK
DDX4
title Germ cell depletion using HSV-TK in mouse testes
title_full Germ cell depletion using HSV-TK in mouse testes
title_fullStr Germ cell depletion using HSV-TK in mouse testes
title_full_unstemmed Germ cell depletion using HSV-TK in mouse testes
title_short Germ cell depletion using HSV-TK in mouse testes
title_sort germ cell depletion using hsv tk in mouse testes
topic Germ cell transplantation
Spermatogonial stem cells
Host mice
Testes
HSV-TK
DDX4
url https://doi.org/10.1038/s41598-025-93325-6
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