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Timing of spermatogonial stem cell transplantation affects the spermatogenic recovery outcome in mice

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Abstract

Spermatogonial stem cell transplantation (SSCT) is a strategy that has demonstrated to be feasible to restore spermatogenesis in animal models when it is performed shortly after the gonadotoxic onset to destroy their endogenous germ cells. However, in the case of boys subjected to fertility preservation, future transplantations will be performed with a delay of many years. In order to study how timing of SSCT affects donor-derived spermatogenic recovery in mice, we compared the percentage of spermatogenic tubule cross-sections within testes of 59 C57BL/6NCrl mice distributed in 6 groups: group 1, untreated mice controls (n = 9); group 2, mice that received a single dose of busulfan 40 mg/kg (n = 10); group 3, mice that received two additional doses of busulfan 10 mg/kg every 5 weeks (n = 10); group 4 (SSCT-A), mice subjected to a standard SSCT performed 5 weeks after a single injection of busulfan 40 mg/kg (n = 10); group 5 (SSCT-B), mice subjected to a delayed SSCT performed 15 weeks after a single injection of busulfan 40 mg/kg (n = 10); and group 6 (SSCT-C), mice subjected to a delayed SSCT with two additional doses of busulfan 10 mg/kg every 5 weeks (n = 10). Spermatogenic recovery in standard SSCT-A and SSCT-C groups ranged between 22.29 and 22.65%, compared with a lower recovery rate of 11.54% showed in the SSCT-B group. However, donor contribution resulted higher in standard SSCT-A, representing a 69.71% of cross-sections, compared with the rest of conditions ranging from 34.69 to 35.42%. Overall, we concluded that a delay in the SSCT from the gonadotoxic onset decreases the efficiency of donor-derived spermatogenic recovery in mice.

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Acknowledgments

We would like to thank all the staff of the animal facility from the Valencia University for their help and patience. We are also grateful to Dr. Isabel Fariñas from the Valencia University for providing us GFP donors for our experiments.

Funding

This work was supported by a private donation of the Celtic Submarí club- Villareal C.F. to Hospital Universitario y Politécnico La Fe intended to promote the scientific research on fertility preservation in child with cancer and an AES project grant (PI16/00931) conceded by the Instituto de Salud Carlos III.

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Authors and Affiliations

  1. Unidad de Medicina Reproductiva, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Av. Fernando Abril Martorell, 106. Torre A, Lab. 6.22, 46026, Valencia, Spain

    J. V. Medrano, A. Navarro-Gomezlechon, A. Pellicer & E. Novella-Maestre

  2. Department of Biology, Faculty of Medicine, Masaryk University, 62500, Brno, Czech Republic

    I. Acimovic

  3. Animal Facility, Faculty of Pharmacy, Valencia University, 46015, Valencia, Spain

    I. Noguera

  4. Fundación IVI, 46026, Valencia, Spain

    A. Pellicer

  5. Hospital Universitario y Politécnico La Fe, 46026, Valencia, Spain

    M. M. Andrés & E. Novella-Maestre

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  1. J. V. Medrano
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  2. I. Acimovic
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  3. A. Navarro-Gomezlechon
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  4. I. Noguera
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  5. A. Pellicer
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  6. M. M. Andrés
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  7. E. Novella-Maestre
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Contributions

JVM, ENM, AP, and MMA conceived this work. JVM, IA, ANG, and IN conducted the experiments. JVM analyzed data and wrote the manuscript. All listed authors revised and approved the manuscript.

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Correspondence to J. V. Medrano.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval

The use of 71 C57BL/6NCrl 5-wk-old male mice as hosts and 12 Tg(CAG-EGFP)B5Nagy/J 3-wk-old males as donors of testicular cells for this study was approved by the Ethical Committee for Animal Welfare of the University of Valencia (ref.: A1513161658035). Mice were housed in purified air-ventilated racks in order to prevent any infection, fed ad libitum, and with controlled dark/light cycle.

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All statistical analyses were performed using SPSS v.25 (IBM).

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Editor: Tetsuji Okamoto

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Medrano, J.V., Acimovic, I., Navarro-Gomezlechon, A. et al. Timing of spermatogonial stem cell transplantation affects the spermatogenic recovery outcome in mice. In Vitro Cell.Dev.Biol.-Animal 57, 21–29 (2021). https://doi.org/10.1007/s11626-020-00531-9

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  • DOI: https://doi.org/10.1007/s11626-020-00531-9

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