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Spermatogenèse in vitro chez la souris : impact sur la qualité nucléaire du spermatozoïde, sur le développement et l'épigénétique de l'embryon issu d'ICSI

Abstract : In recent years, testicular biopsy followed by the freezing of testicular tissue has been proposed to children with cancer before the introduction of a gonadotoxic treatment. This fertility preservation procedure is offered with the hope that a fertility restoration method will be developed. The thawed testicular tissue could thus be used to perform in vitro maturation, avoiding the reintroduction of tumor cells, to produce spermatozoa. This thesis work first consisted in assessing the establishment of DNA methylation in mouse prepubertal testicular tissue during in vitro spermatogenesis. The culture of fresh or thawed mouse testicular testicular tissue allows the expression levels of DNA methyltransferases 1 and 3a to be maintained in spermatogonia and spermatocytes. In addition, DNA methylation is found even in in vitro produced spermatozoa. The nuclear quality of these spermatozoa was then analyzed. The culture of testicular tissue has no impact on sperm aneuploidy rate, chromatin condensation and DNA fragmentation. However, freezing followed by organotypic culture increases the proportion of spermatozoa with oxidized DNA. Finally, the functionality of in vitro produced spermatozoa was analyzed by oocyte microinjection and the dynamics of different epigenetic marks was studied during preimplantation development. Embryo developmental rates are decreased when using in vitro produced spermatozoa. The levels of H3K4me3, H3K27me3 and H3K9ac are slightly modified in embryos derived from spermatozoa generated in vitro whereas DNA methylation and demethylation are more affected. The production of spermatozoa after culture of fresh or thawed prepubertal tissue in the mouse model has shown that this procedure is not without impact on the early embryo, although the quality of the spermatozoa produced is relatively unaltered.
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https://tel.archives-ouvertes.fr/tel-03196600
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Submitted on : Tuesday, April 13, 2021 - 1:12:00 AM
Last modification on : Friday, May 7, 2021 - 3:19:12 AM

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  • HAL Id : tel-03196600, version 1

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Antoine Oblette. Spermatogenèse in vitro chez la souris : impact sur la qualité nucléaire du spermatozoïde, sur le développement et l'épigénétique de l'embryon issu d'ICSI. Biologie de la reproduction. Normandie Université, 2019. Français. ⟨NNT : 2019NORMR004⟩. ⟨tel-03196600⟩

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