Human testis in organotypic culture: application for basic or clinical research

Roulet, Vanessa; Denis, Hélène; Staub, Christophe; Tortorec, Anna Le; Delaleu, Bernadette; Satie, Anne‐Pascale; Patard, Jean‐Jacques; Jégou, Bernard; Dejucq-Rainsford, Nathalie

doi:10.1093/humrep/del018

Cited by 76 publications

(70 citation statements)

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“…The observation by transillumination allowed us to ascertain that the testes displayed spermatogenesis (Roulet et al, 2006). Four 3-mm 3 testis explants were placed onto a PET insert (Falcon Labware; Becton Dickinson, Lincoln Park, NJ, USA) at the interface of air in 1 mL of Dulbecco's Modified Eagle's Medium supplemented with antibiotics, 1 mM sodium pyruvate, 4 mM glutamine, 100 ng/ml of vitamin A, 200 ng/ml of vitamin E, 50 ng/ml of vitamin C, 10 µg/ml of insulin and 5 µg/ml of transferrin and with 1 IU/ml hCG for culture, in 12-well plates.…”

Section: The Testis Explant Assay (Texas)mentioning

confidence: 99%

Parallel assessment of the effects of bisphenol A and several of its analogs on the adult human testis

et al. 2017

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WHAT IS KNOWN ALREADY:The few epidemiologic studies performed suggest that bisphenols have potential endocrine disruptive properties, but they did not identify clear and direct patterns of endocrine disruption. STUDY DESIGN, SIZE, DURATION:Adult human testis explants in culture were exposed to BPA and the analogs bisphenol F (BPF), bisphenol S (BPS), bisphenol E (BPE), bisphenol B (BPB), and bisphenol A diglycidyl ether (BADGE) at 10 -9 to 10 -5 M for 24 h or 48h.PARTICIPANTS/MATERIALS, SETTING, METHODS: Human adult testes were obtained from prostate cancer patients who had no hormone therapy, or from multiorgan donors. After ex vivo exposure to the investigated bisphenols, the measured outcomes were related to histopathology (gross morphology and germ cell viability determined by anti-caspase 3 immunohistochemistry), and the levels of testosterone, insulin-like fator 3 and inhibin B were measured using immunoassays. The levels of mRNA encoding key enzymes of bisphenol biotransformation were investigated by quantitative PCR: UGT2B15 UDP (glucuronosyltransferase two family, polypeptide B15), GUSB (glucuronidase beta), SULT1A1 and 3 (sulfotransferase family 1 A member 1 and 3) and STS (steroid sulfatase). MAIN RESULTS AND THE ROLE OF CHANCE: A significant dose-dependent inhibition wasfound between testosterone levels measured in the culture medium and concentrations of BPA The active concentrations of BPA and BPA-A used in the study were higher than those found in human biological fluids. WIDER IMPLICATIONS OF THE FINDINGS:Under our experimental conditions, direct exposure to BPA or BPA-A can result in endocrine disturbance in the adult human testis.

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Section: The Testis Explant Assay (Texas)mentioning

confidence: 99%

Parallel assessment of the effects of bisphenol A and several of its analogs on the adult human testis

et al. 2017

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“…The materials were assessed by translumination for the occurrence of spermatogenesis before their subsequent use, and culture was performed as previously described. 24 In brief, testicular tissue was dissected into 3-mm 3 fragments and two explants transferred onto a polyethylene terephthalate insert in a well of a six-well plate (Falcon Labware; Becton-Dickinson and Co., Lincoln Park, NJ) containing 2 ml of media (Dulbecco's modified Eagle's medium supplemented with antibiotics, 10% fetal calf serum, 1 mmol/L sodium pyruvate, 4 mmol/L glutamine, 100 ng/ml vitamin A, 200 ng/ml vitamin E, 50 ng/ml vitamin C, 10 g/ml insulin, and 5 g/ml transferrin). The culture was established for 14 to 16 days in a humidified atmosphere containing 5% CO 2 at 37°C, medium changed every 2 days, and stored frozen at Ϫ20°C.…”

Section: Organotypic Culture Of Human Testis Explantsmentioning

confidence: 99%

“…As previously reported, adult human testis explants in culture for up to 2 weeks retained overall structural integrity of both the interstitial and the seminiferous tubules compartments, and all germ cell types associated with Sertoli cells presented classic ultrastructural characteristics, testosterone secretion, and expression of somatic cell markers. 24 Every 2 days, testis explants were either fixed in neutral buffered 4% formaldehyde for immunohistochemical analysis or frozen and stored at Ϫ80°C until nucleic acid extraction.…”

Section: Organotypic Culture Of Human Testis Explantsmentioning

confidence: 99%

“…14 -23 We have recently developed an organotypic culture of human testis 24 with the primary objective of studying HIV infection within a control environment preserving cellular interactions. Using this in vitro model, and based on a series of experiments in situ, the present study provides a detailed analysis of HIV receptor expression within the testis and investigates the ability of this organ to be infected by a dual-tropic HIV-1 strain in vitro.…”

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Susceptibility of Human Testis to Human Immunodeficiency Virus-1 Infection in Situ and in Vitro

Roulet

Satie

Ruffault

et al. 2006

The American Journal of Pathology

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Semen represents the main vector for human immunodeficiency virus (HIV) dissemination worldwide and has been shown to harbor replication-competent virus despite otherwise effective highly active anti-retroviral therapy, which achieves undetectable viral load in plasma. Despite this, the origin of seminal HIV particles remains unclear, as does the question of whether the male genital tract organs contribute virus to semen. Here we investigated the presence of HIV receptors within the human testis using immunohistochemistry and quantitative real-time polymerase chain reaction. We also analyzed the infectivity of a dual tropic HIV-1 strain in an organotypic culture, as well as the impact of viral exposure on testosterone production. Our study establishes that CXCR4 ؉ , CCR5 ؉ , CD4 ؉ , and DC-SIGN With sexual contact being the main cause of the spread of human immunodeficiency virus (HIV) and male to female transmission rates being higher and more efficient than female to male, semen represents the foremost vector of HIV dissemination worldwide. However, the origin of the virus in the semen is still unclear. Several arguments point to the existence of local sources producing free viral particles in this bodily fluid. First, a number of studies clearly indicate that semen represents a viral compartment distinct from the blood.

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“…Avant nos travaux, peu d'études avaient été effectuées sur l'infection du tractus génital masculin en raison de la difficulté d'accès aux organes génitaux d'hommes VIH + . Afin de pallier ce problème, notre équipe a développé une culture organotypique de tissu testiculaire [30] qui a permis de montrer que le testicule humain est infecté par le VIH-1 in vitro [31]. In vivo, l'infection de cet organe a été mise en évidence dans un modèle animal qui mime l'infection chez l'homme -les macaques cynomolgus infectés par le SIV (virus de l'immunodéficience simienne).…”

Section: Organes Du Tractus Génital Masculin Et Vihunclassified

Le tractus génital masculin

Tortorec

Dejucq-Rainsford

2010

Med Sci (Paris)

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Peu après l'apparition des premiers cas de Sida en 1981 et avant même la découverte de son agent étiologique le virus de l'immunodéficience humaine (VIH), il a été postulé que la cause du Sida était un agent pathogène transmis par voie sexuelle et sanguine. Presque trente ans plus tard, le VIH s'est disséminé à l'ensemble de la planète et a infecté plus de 65 millions de personnes. Plus de 80 % des infections se font par voie sexuelle. Le sperme représente le principal vecteur de dissémination du VIH, la transmission étant plus efficace de l'homme à la femme (environ 50 % des personnes infectées par le VIH dans le monde sont des femmes à l'heure actuelle) ou de l'homme à l'homme que de la femme à l'homme. Paradoxalement, l'origine des particules virales libres et des cellules infectées présentes dans le sperme demeure très mal connue. Le sperme est constitué de cellules (parmi lesquelles les spermatozoïdes, les leucocytes, les cellules germinales immatures) et de sécré-tions (le liquide séminal) provenant des testicules, des épididymes, de la prostate, des vésicules séminales et des glandes bulbo-urétrales. L'infection de ces organes par le VIH a fait l'objet de peu d'études en raison de la difficulté d'accès aux organes reproducteurs d'hommes séropositifs pour le VIH. De ce fait, plusieurs questions essentielles restent posées (Figure 1) : -Quelle est la nature des sources productrices de virus dans le tractus ? Il est établi que les souches virales spermatiques ne proviennent pas uniquement du compartiment sanguin. Un ou plusieurs organes de l'appareil reproducteur sont-ils responsables de la charge virale spermatique ? -Quel est l'impact des protéines/peptides du sperme infecté sur la transmission sexuelle ? -Le tractus génital constitue-t-il un réservoir viral résistant aux traitements antirétroviraux (ou ART pour antiretroviral therapy) ? De nombreuses études ont montré que les thérapies anti-VIH ne permettent pas toujours d'éliminer la contamination du sperme par le VIH, même lorsque la virémie est indétectable. -Quelle est la cause des altérations du sperme observées chez des hommes porteurs du VIH et asymptomatiques ?> Alors que le sperme représente le principal vecteur de dissémination du virus de l'immunodéficience humaine (VIH) dans le monde, l'origine du virus dans ce fluide demeure très mal connue. Chez certains hommes sous traitement antirétroviral efficace depuis plusieurs mois voire plusieurs années, le VIH persiste dans le sperme. Il est donc essentiel de déterminer la nature des sources de virus à ce niveau pour améliorer l'efficacité des thérapies actuelles. Nos travaux ont permis de mettre en évidence la réplication du VIH dans les organes participant à l'élaboration du sperme, confortant les analyses phylogénétiques qui indiquent que les particules virales et les cellules infectées présentes dans le sperme sont en partie produites localement. Cette production de virus au sein du tractus génital masculin pose de nombreuses questions : l'infection du testicule et/ou des glandes annexes est-ell...

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Human testis in organotypic culture: application for basic or clinical research

Abstract: Although this culture model is clearly unsuitable for preparing germ cells for therapeutic purposes, it does represent a most valuable tool for testing the effects of biological and chemical agents on testicular tissue.

Cited by 76 publications

References 47 publications

Parallel assessment of the effects of bisphenol A and several of its analogs on the adult human testis

Parallel assessment of the effects of bisphenol A and several of its analogs on the adult human testis

Susceptibility of Human Testis to Human Immunodeficiency Virus-1 Infection in Situ and in Vitro

Le tractus génital masculin

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