Assessment of sperm nuclear quality after in vitro maturation of fresh or frozen/thawed mouse pre-pubertal testes

Oblette, Antoine; Rives, Nathalie; Dumont, Ludovic; Rives, A; Verhaeghe, France; Jumeau, Fanny; Rondanino, Christine

doi:10.1093/molehr/gax048

Cited by 21 publications

(21 citation statements)

References 39 publications

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“…Thus, in vitro spermatogenesis appears to be the most appropriate approach to avoid the potential risk of reintroducing tumour cells in the cured patient. The three-dimensional cell culture and the organotypic culture have been shown to support the differentiation of spermatogonia into spermatozoa in the rat and mouse model [156][157][158][159]. Moreover, the generation of sperm-like cells has been reported in 3D cell culture from spermatogonial cells of busulfan-treated mice [160].…”

Section: Discussionmentioning

confidence: 99%

Exposure to Chemotherapy During Childhood or Adulthood and Consequences on Spermatogenesis and Male Fertility

Delessard

Saulnier

Rives

et al. 2020

IJMS

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Over the last decade, the number of cancer survivors has increased thanks to progress in diagnosis and treatment. Cancer treatments are often accompanied by adverse side effects depending on the age of the patient, the type of cancer, the treatment regimen, and the doses. The testicular tissue is very sensitive to chemotherapy and radiotherapy. This review will summarize the epidemiological and experimental data concerning the consequences of exposure to chemotherapy during the prepubertal period or adulthood on spermatogenic progression, sperm production, sperm nuclear quality, and the health of the offspring. Studies concerning the gonadotoxicity of anticancer drugs in adult survivors of childhood cancer are still limited compared with those concerning the effects of chemotherapy exposure during adulthood. In humans, it is difficult to evaluate exactly the toxicity of chemotherapeutic agents because cancer treatments often combine chemotherapy and radiotherapy. Thus, it is important to undertake experimental studies in animal models in order to define the mechanism involved in the drug gonadotoxicity and to assess the effects of their administration alone or in combination on immature and mature testis. These data will help to better inform cancer patients after recovery about the risks of chemotherapy for their future fertility and to propose fertility preservation options.

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Section: Discussionmentioning

confidence: 99%

Exposure to Chemotherapy During Childhood or Adulthood and Consequences on Spermatogenesis and Male Fertility

Delessard

Saulnier

Rives

et al. 2020

IJMS

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“…Indeed, seminiferous tubules are directly exposed to air in the gas-liquid interphase culture system. Nonetheless, we recently showed that no increased generation of spermatozoa containing 8-OHdG adducts was found in cultures of fresh tissues supplemented with Vit E [60]. Therefore, it can be suggested that the structural integrity of post-thaw testicular cell membranes is better preserved in the presence of Vit E that facilitates inter-communication between the different cell types necessary for the good course of spermatogenesis during in vitro culture of the F-T testicular tissue.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, it can be suggested that the structural integrity of post-thaw testicular cell membranes is better preserved in the presence of Vit E that facilitates inter-communication between the different cell types necessary for the good course of spermatogenesis during in vitro culture of the F-T testicular tissue. However, more spermatozoa presented oxidative damages in cultures of F-T tissues, suggesting that cryopreservation procedures may alter the defensive capacity of undifferentiated and differentiated germ cells against oxidative stress [60].…”

Section: Discussionmentioning

confidence: 99%

Vitamin E but Not GSH Decreases Reactive Oxygen Species Accumulation and Enhances Sperm Production during In Vitro Maturation of Frozen-Thawed Prepubertal Mouse Testicular Tissue

Arkoun

Galas

Dumont

et al. 2019

IJMS

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Freezing–thawing procedures and in vitro culture conditions are considered as a source of stress associated with increased reactive oxygen species (ROS) generation, leading to a damaged cell aerobic metabolism and consequently to oxidative stress. In the present study, we sought to investigate whether vitamin E (Vit E) or reduced glutathione (GSH) enhances sperm production by decreasing ROS accumulation during in vitro maturation of prepubertal mice testes. Testes of prepubertal mice were cryopreserved using a freezing medium supplemented or not supplemented with Vit E and were cultured after thawing. In presence of Rol alone in culture medium, frozen-thawed (F-T) testicular tissues exhibited a higher ROS accumulation than fresh tissue during in vitro culture. However, Vit E supplementation in freezing, thawing, and culture media significantly decreased cytoplasmic ROS accumulation in F-T testicular tissue during in vitro maturation when compared with F-T testicular tissue cultured in the presence of Rol alone, whereas GSH supplementation in culture medium significantly increased ROS accumulation associated with cytolysis and tissue disintegration. Vit E but not GSH promoted a better in vitro sperm production and was a suitable ROS scavenger and effective molecule to improve the yield of in vitro spermatogenesis from F-T prepubertal mice testes. The prevention of oxidative stress in the cytoplasmic compartment should be regarded as a potential strategy for improving testicular tissue viability and functionality during the freeze–thaw procedure and in vitro maturation.

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“…However, the spermatogenic yield remains low in in vitro‐matured tissues compared with the in vivo situation, with a decreased meiotic and post‐meiotic progression and an increased formation of degenerating round spermatids 6,12 . However, our recent data showed that the nuclear quality of in vitro‐produced spermatozoa from fresh or frozen/thawed testicular tissues was similar to that of their in vivo counterparts 10 …”

Section: Introductionmentioning

confidence: 86%

Dynamics of epigenetic modifications in ICSI embryos from in vitro‐produced spermatozoa

et al. 2020

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Background In prepubertal boys with cancer, fertility preservation relies on testicular tissue freezing before treatment. In vitro maturation of frozen/thawed tissues could be one of the procedures envisaged to restore the fertility of cured patients. It is necessary to ascertain in the mouse model that in vitro‐generated spermatozoa are able to ensure embryo development, without altering the epigenetic processes occurring during the pre‐implantation period. Objectives The aims of the present study were to investigate the fertilizing ability of in vitro‐produced spermatozoa and explore several epigenetic marks at different stages of embryo development. Materials and methods Fresh or controlled slow‐frozen (CSF)/thawed testicular tissues from 6 to 7 days post‐partum (dpp) mice were cultured for 30 days. Intracytoplasmic sperm injection (ICSI) experiments were performed using in vitro‐produced spermatozoa. Testicular spermatozoa from 36 to 37 dpp mice were used as in vivo controls. DNA methylation/hydroxymethylation and histone post‐translational modifications (H3K4me3, H3K27me3 and H3K9ac) were analysed by immunofluorescence from the zygote to the blastocyst stages. Results The spermatozoa generated in cultures of fresh or CSF testicular tissues were able to initiate embryonic development. The freezing of prepubertal testicular tissues limits the production of spermatozoa in vitro and the fertilization rate after ICSI. Similar levels of H3K4me3, H3K27me3 and H3K9ac were found in ICSI embryos derived from in vitro‐ and in vivo‐produced spermatozoa. DNA methylation levels were increased in 4‐cell embryos and morula obtained by ICSI with in vitro‐produced spermatozoa. Discussion and conclusion Our study shows for the first time that the use of in vitro‐produced spermatozoa alters DNA methylation/demethylation dynamics but has little impact on H3K4me3, H3K27me3 and H3K9ac levels in mouse early embryos. Further work will have to be performed to determine whether the use of these gametes is not deleterious for embryo development before considering a human application.

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Assessment of sperm nuclear quality after in vitro maturation of fresh or frozen/thawed mouse pre-pubertal testes

Cited by 21 publications

References 39 publications

Exposure to Chemotherapy During Childhood or Adulthood and Consequences on Spermatogenesis and Male Fertility

Exposure to Chemotherapy During Childhood or Adulthood and Consequences on Spermatogenesis and Male Fertility

Vitamin E but Not GSH Decreases Reactive Oxygen Species Accumulation and Enhances Sperm Production during In Vitro Maturation of Frozen-Thawed Prepubertal Mouse Testicular Tissue

Dynamics of epigenetic modifications in ICSI embryos from in vitro‐produced spermatozoa

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