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Study question What is the proportion of men treated for testicular cancer or lymphoma who banked sperm before gonadotoxic treatment in France? Summary answer In 2018, 41% of men with testicular cancer and 28% of men with lymphoma banked sperm before gonadotoxic treatment in France. What is known already Cancer treatments significantly improved survival rates in men. In Europe, the 5-year survival rate is above 90% for testicular cancer (TC) and Hodgkin lymphoma (HL), which, with Non Hodgkin Lymphoma (NHL), are the most frequent cancers in men of reproductive age. However, surgery, chemotherapy and/or radiotherapy may damage male fertility: between 15% and 30% of men remain permanently infertile after treatment. Since 2011, the French bioethics law recommends and guarantees free access to fertility preservation for all patients needing gonadotoxic treatments. Few international data are available about the rate of male cancer patients who receive sperm cryopreservation before cancer treatment. Study design, size, duration We analyzed the data of male cancer incidence in 2018 published by the French National Institute of Cancer (INCa), only available and latest French national data, estimated from cancer registers of selected metropolitan departments (Defossez et al. 2019). We organized a survey among the 27 metropolitan centers of the CECOS network (public centers for study and conservation of human eggs and sperm) to collect the annual number of sperm cryopreservations for TC, HL or NHL. Participants/materials, setting, methods We compared the number of 0-59 years-old men with TC, HL or NHL in metropolitan France (extracted from 2018 INCa data) and the number of sperm banking for TC, HL or NHL in metropolitan centers of the CECOS network. Main results and the role of chance INCa estimated that 38,048 new cancers were diagnosed in metropolitan France in 2018 in men aged 0-59 years. TC accounted for 2,630 new cases and lymphomas for 3,913 new cases (943 HL and 2,970 NHL). Twenty-six out of 27 metropolitan CECOS centers answered the survey (96% participation rate): in 2018, 1,079 men banked sperm for TC, 375 for HL and 211 for NHL. In 2018, the fertility preservation rate in metropolitan France was 41% for TC and 28% for lymphomas (51% for HL, and 15% for NHL). The results of our national study are in accordance with Uçar et al. 2020, a monocentric study on 110 TC patients showing a 43% fertility preservation rate, but differ from those of Abdel-Razek et al. 2020, a monocentric study on 59 NHL and 26 HL patients, showing respectively 32% and 14% fertility preservation rates. A limit of our study is to be based on estimated cancer incidence from INCa reports without stratification on age and parenthood; sperm banking activity was measured in the CECOS network, which performs 90% of French male fertility preservation (French Biomedicine Agency 2018 data). This design may induce an underestimation of cancer incidence and of sperm banking activity. Limitations, reasons for caution Our study did not consider patients who were informed of fertility preservation but did not want to/could not bank ejaculated sperm (azoospermia, sperm collection failure, no-show). In CECOS network, the estimated rate of such patients is 10%. French overseas regions were not studied (lack of INCa data). Wider implications of the findings Our results suggest that despite the recommendations and facilities offered by the French bioethics law, the male patients’ pathway for fertility preservation before cancer treatments could be improved. Further study should evaluate if this suboptimal rate of fertility preservation is homogeneous on French territory or related to postal code. Trial registration number Not applicable

Study question What is the impact of the cancer treatment received prior to TTF and the disease on spermatogonia quantity in testicular tissue from (pre)pubertal boys? Summary answer A decrease in spermatogonia number was observed in testicular tissue after cancer treatment when cyclophosphamide equivalente dose (CED) is above 4000 mg/m2. What is known already The improved survival rates associated with the development of sperm and testicular tissue freezing (TTF) renders difficult not to offer fertility preservation to children or adolescents before cancer. Several studies exploring cancer patients have examined the number of spermatogonia per seminiferous tubular cross-section (S/T) or tubular fertility index (TFI, percentage of tubular cross-sections containing spermatogonia) in testicular biopsies. All studies, demonstrated that the S/T and TFI always decreased after the introduction of chemotherapy and more specifically in case of highly gonadotoxic risk such as alkylating agents. Study design, size, duration Testicular tissue samples from 79 (pre)pubertal boys diagnosed with cancer (ranging from 6 month to 16 years of age) were cryopreserved between May 2009 and June 2014. Medical diagnosis and previous chemotherapy exposure were recorded. We examined histological sections of (pre)pubertal testicular tissue to elucidate whether chemotherapy, doses or primary diagnosis affects the quality of testicular tissue. Participants/materials, setting, methods (Pre)pubertal boys with cancer diagnosis who benefitted from TTF prior to conditioning treatment for hematopoietic stem cell transplantation. All the patients included had previously received chemotherapy with moderate risk for future fertility. We have selected patients for whom data on chemotherapy received were complete. The quantity of spermatogonia and quality of testicular tissue were assessed by both morphological and immunohistochemical analysis. Main results and the role of chance The main finding was a significant reduction in spermatogonial cell counts in boys treated with alkylating agents. The mean S/T values in boys exposed to alkylating agents was significantly lower than in a group exposed to non-alkylating agents (p = 0.018). In contrast, no difference was observed for patients treated with carboplatin as the only alkylating agent compared to the group of patients exposed to non-alkylating agents. We observed an increase of S/T with age in the group of patients who did not receive alkylating agents and a decrease of S/T with age when patients received alkylating agents included in the CED formula (r = 0.6166, p = 0.0434; r= -0.3759, p = 0.0036, respectively). The TFI and S/T were decreased in the group of patients who received vincristine (p = 0.0049; p Limitations, reasons for caution This is a descriptive study of testicular tissues obtained from patients who were at risk of infertility. Spermatognia functionanlity could not be tested by transplantation due limited sample size. Wider implications of the findings This study summarizes spermatogonia quantity and quality of testicular tissue of (pre)pubertal boys after potentially sterilizing treatments. We confirmed a negative correlation between the cumulative exposure to alkylating agents and the spermatogonial quantity. For patients in whom fertility preservation is indicated, TTF should be performed before initiation of alkylating agents. Trial registration number N/A

STUDY QUESTION In a non-commercial national gamete donation programme, do the motivations and personality characteristics of candidate sperm and oocyte donors differ according to their parenthood status? SUMMARY ANSWER Moderate differences exist between non-parent and parent candidate donors in motivations for gamete donation and representations as well as in personality characteristics. WHAT IS KNOWN ALREADY Several studies have analysed the motivations and experiences of oocyte or sperm donors, but mainly in countries where gamete donation is a commercial transaction, and very few studies have reported results of personality traits using personality inventory tests. No study has specifically investigated the motivations and personality characteristics of candidate gamete donors according to parenthood status. STUDY DESIGN, SIZE, DURATION A prospective study was carried out including 1021 candidate donors from 21 centres (in university hospitals) of the national sperm and egg banking network in France between November 2016 and December 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS In total, 1021 candidate gamete donors were included in the study. During their first visit, male (n = 488) and female candidate donors (n = 533) completed a questionnaire on sociodemographic characteristics, their motivations for donation and their representations of donation, infertility and family. Secondly, a NEO Personality Inventory (NEO-PI-R) exploring the Big Five personality traits was completed online. Results were compared between parent and non-parent candidate donors. MAIN RESULTS AND THE ROLE OF CHANCE Altruistic values were the principal motive for donation irrespective of parenthood status. Reassurance about their fertility or preservation of sperm for future use was more often reported in non-parent than in parent candidate donors. With regard to representation of gamete donation or of the family, independently of their parenthood status, candidate donors more frequently selected social rather than biological representations. Mean personality characteristics were in the normal range. Non-parent candidate donors had higher scores on openness and depression than parents, while parent candidate donors appeared more social than non-parents. LIMITATIONS, REASONS FOR CAUTION The personality characteristics inventory was not completed by all candidate donors included in the study. However, family status did not differ between the two groups (NEO-PI-R completed (n = 525) or not), while the group who completed the NEO-PI-R had a higher educational level. This national study was performed in a country where gamete donation is subject to strict legislation. WIDER IMPLICATIONS OF THE FINDINGS In a global context where reproductive medicine is commercialized and gamete donor resources are limited, this study found that altruism and social representations of gamete donation and family are the main motivations for gamete donation in a country which prohibits financial incentive. These findings are relevant for health policy and for gamete donation information campaigns. STUDY FUNDING/COMPETING INTEREST(S) Grant from the Agence de la Biomédecine, France. The authors have nothing to disclose related to this study. TRIAL REGISTRATION NUMBER N/A.

Fertility preservation is essential before cancer treatment. When ejaculated sperm preservation is not possible, testicular tissue can be surgically collected by Onco-TESE technic (Oncological Testicular Sperm Extraction) to isolate sperm. We report on our experience with Onco-TESE in testicular cancer patients at the Rouen University Hospital.Retrospective study including all pubescent men, treated for testicular cancer, uni- or bilateral, before any carcinological therapy, who have undergone Onco-TESE at the Rouen University Hospital. Fragment weight, detection of sperm or its precursors were analysed. A histological interpretation of the testicular tumor was carried out. For each positive sample, straws were kept at the French Sperm Bank.Twenty-four patients had an Onco-TESE: 58.34% severe sperm alteration (SSA) and 41.36% sperm collection failure (SCF), between 1996 and 2019. The mean age was 26.6 (±5.29) years. The mean procedure and length of stay were 71minutes (±30.7) and 3.75 days (±2.83), respectively. The rate of positive testicular biopsies (TB) was 58.33% overall and 66,67% in the case of TB on tumour testis. One patient had a Clavian-Dindo III complication. The mean number of straws preserved per patient was 14.28 (±15.34) for 7.14% use.Our results seem to confirm that Onco-TESE is an effective solution for preserving fertility in men with testicular cancer in cases of SSA or SCF.III.

RESUME La conservation des ovocytes s’effectue par vitrification, en France depuis 2011, et seules y sont autorisees les conservations destinees a la preservation de la fertilite, donc les indications medicales. Cette technique s'adresse surtout aux femmes atteintes de pathologies malignes, dont un traitement gonadotoxique, par chimiotherapie ou radiotherapie risque de degrader la fonction ovarienne. Elle concerne pareillement les femmes menacees d'insuffisance ovarienne prematuree. A ces indications se sont ajoutees depuis quelques annees des demandes dites « non medicales », en fait de palliation de l'infertilite liee a l'avancee en âge. Sans perspective previsible de grossesse, surtout faute de stabilite de couple, des femmes sensibles a la baisse de la fertilite avec l'âge, souhaitent faire prelever et conserver leurs ovocytes afin d’y avoir eventuellement recours plus tard, par FIV, si elles eprouvaient alors des difficultes a concevoir. Les chances de grossesse dependent essentiellement de l'âge auquel les ovocytes ont ete recueillis, au mieux avant 35 ans. Les risques et les avantages possibles de ces demarches, sont discutes. Non autorisees en France elles sont effectuees par les femmes francaises dans les centres etrangers. Cependant la Loi de Bioethique du 07 juillet 2011 a etendu aux femmes majeures n'ayant jamais procree la possibilite de participer au don d’ovocytes. En contrepartie ces femmes peuvent conserver pour elles-memes, si leur nombre le permet, une partie des ovocytes recueillis. Ainsi est ouverte la voie legale de la conservation des ovocytes pour palliation de l'infertilite liee a l'âge.

Resume Objectifs L’amelioration des connaissances sur la gonadotoxicite et le caractere peu invasif de la conservation de spermatozoides, ont permis d’accroitre les indications de preservation de la fertilite. Le but de ce travail est de preciser ces indications et les techniques de conservation, afin d’informer et sensibiliser les medecins susceptibles d’adresser des patients au CECOS. Methodes Une revue de la litterature a ete realisee sur la base de recherche PubMed portant sur les indications et techniques de preservation de la fertilite chez l’homme pubere et prepubere. Resultats Avant toute chimio et/ou radiotherapie et/ou chirurgie, il est indispensable d’evaluer le risque gonadotoxique ou de troubles de l’erection et/ou d’ejaculation secondaire, afin d’informer et adresser le patient pour preservation de sa fertilite. Conclusion Il vaut mieux congeler par exces, d’autant plus que le developpement constant de nouvelles molecules laisse peu de recul pour evaluer leur risque gonadotoxique.

BACKGROUNDInfertility is an important side effect of treatments used for cancer and other non-malignant conditions in males. This may be due to the loss of spermatogonial stem cells (SSCs) and/or altered functionality of testicular somatic cells (e.g. Sertoli cells, Leydig cells). Whereas sperm cryopreservation is the first-line procedure to preserve fertility in post-pubertal males, this option does not exist for prepubertal boys. For patients unable to produce sperm and at high risk of losing their fertility, testicular tissue freezing is now proposed as an alternative experimental option to safeguard their fertility.OBJECTIVE AND RATIONALEWith this review, we aim to provide an update on clinical practices and experimental methods, as well as to describe patient management inclusion strategies used to preserve and restore the fertility of prepubertal boys at high risk of fertility loss.SEARCH METHODSBased on the expertise of the participating centres and a literature search of the progress in clinical practices, patient management strategies and experimental methods used to preserve and restore the fertility of prepubertal boys at high risk of fertility loss were identified. In addition, a survey was conducted amongst European and North American centres/networks that have published papers on their testicular tissue banking activity.OUTCOMESSince the first publication on murine SSC transplantation in 1994, remarkable progress has been made towards clinical application: cryopreservation protocols for testicular tissue have been developed in animal models and are now offered to patients in clinics as a still experimental procedure. Transplantation methods have been adapted for human testis, and the efficiency and safety of the technique are being evaluated in mouse and primate models. However, important practical, medical and ethical issues must be resolved before fertility restoration can be applied in the clinic.Since the previous survey conducted in 2012, the implementation of testicular tissue cryopreservation as a means to preserve the fertility of prepubertal boys has increased. Data have been collected from 24 co-ordinating centres worldwide, which are actively offering testis tissue cryobanking to safeguard the future fertility of boys. More than 1033 young patients (age range 3 months to 18 years) have already undergone testicular tissue retrieval and storage for fertility preservation.LIMITATIONS, REASONS FOR CAUTIONThe review does not include the data of all reproductive centres worldwide. Other centres might be offering testicular tissue cryopreservation. Therefore, the numbers might be not representative for the entire field in reproductive medicine and biology worldwide. The key ethical issue regarding fertility preservation in prepubertal boys remains the experimental nature of the intervention.WIDER IMPLICATIONSThe revised procedures can be implemented by the multi-disciplinary teams offering and/or developing treatment strategies to preserve the fertility of prepubertal boys who have a high risk of fertility loss.STUDY FUNDING/COMPETING INTEREST(S)The work was funded by ESHRE. None of the authors has a conflict of interest.

Resume Introduction Depuis la loi du 4 juillet 2001, la vasectomie est reconnue comme methode de contraception masculine. Nous rapportons l’experience de la pratique de vasectomie au sein d’un centre hospitalo-universitaire. Materiels et methodes Etude de cohorte retrospective monocentrique de 45 patients ayant beneficie d’une vasectomie contraceptive entre juillet 2001 et mai 2016. Pour chaque patient ont ete etudie : les modalites de realisation, le respect des recommandations issues de la loi de 2001, les couts et benefices engendres par l’intervention, l’efficacite du geste sur les spermogrammes de controle, la satisfaction des patients par un questionnaire telephonique. Resultats L’âge moyen etait de 41,3 ans. La seconde consultation etait realisee dans 91 % des cas mais le delai de reflexion non respecte dans 24 % des cas. Le consentement ecrit etait signe dans 89 % des cas. La vasectomie etait realisee en ambulatoire dans 73 % des cas, sous anesthesie locale dans 6,7 % des cas. Le cout moyen par patient etait de 660,63 euros pour un gain moyen de 524,50 euros soit une perte de 136,13 euros. Sur le spermogramme de controle, 54,3 % etaient azoospermes mais le delai de 3 mois n’etait pas respecte chez 23 % d’entre eux. Aucun patient n’a exprime de regret apres l’intervention. Conclusion Les recommandations issues de la loi de 2001 n’etaient pas systematiquement suivies. Ce manque de standardisation des pratiques, potentiel reflet d’un manque d’interet, est a mettre en relief avec le surcout engendre. La revalorisation de l’acte devrait etre integree dans la reflexion d’amelioration des pratiques de sterilisation masculine. Niveau de preuve 4.

Objectifs La preservation de la fertilite est indispensable avant tout traitement pour cancer, en particulier chez les hommes jeunes. En cas d’impossibilite de recueil de gametes, un prelevement chirurgical de pulpe testiculaire est toujours possible dans le cadre d’une onco-tese (testicular sperm extraction). Nous rapportons notre experience d’onco-tese. Methodes Etude retrospective incluant l’ensemble des patients masculins puberes avant tout traitement pour un cancer (orchidectomie, chimiotherapie, radiotherapie ou chirurgie pouvant alterer l’ejaculation) et ayant eu une onco-tese. Les biopsies etaient realisees par abord scrotal sous anesthesie generale, puis envoyees en biologie de la reproduction et en anatomopathologie. Le poids des fragments, la presence de spermatozoides, de spermatogonies, de spermatocytes et de spermatides, le pourcentage de spermatozoides mobiles avant et apres congelation et la realisation ou non d’un test de mobilite ont ete analyses. Pour chaque prelevement positif, des paillettes ont ete conservees au cecos. Resultats Quarante-deux patients ont eu une onco-tese, dans notre centre entre 1996 et 2017 (59,52 % pour alteration du spermogramme et 40,48 % pour echec de recueil). L’âge moyen etait de 27 ans. Le cancer sous-jacent etait : tumeur du testicule 64,29 %, hemopathies malignes 19,05 %, sarcomes osseux 7,14 %, autres 9,52 %. Le delai median entre la proposition et l’intervention etait de 5 jours Pour 69,05 % des patients, la biopsie etait associee a un geste necessaire pour le traitement du cancer. Seize patients ont eu un prelevement unilateral. La duree moyenne d’hospitalisation etait de 3 jours et 1 patient a presente une complication Clavien I. Le taux de biopsies positives etait de 61,90 %. Le nombre moyen de paillettes conservees par patient etait de 9,52. Au total, 14,28 % des patients ont eu egalement une conservation de pulpe testiculaire ( Tableau 1 ). Conclusion A notre connaissance, cette serie est la plus importante rapportee sur ce sujet. Nos resultats semblent confirmer que l’onco-tese est une solution efficace pour preserver la fertilite chez les hommes jeunes atteints de cancer en cas d’echec au recueil ou d’alteration majeure du spermogramme. C’est une technique fiable, accessible en urgence, qui devrait etre proposee systematiquement.

Intracytoplasmic morphologically selected sperm injection (IMSI), by selecting spermatozoa at high magnification improves the outcome of intracytoplasmic sperm injection (ICSI) mainly after several failures. However, only few monocentric randomized studies are available and they do not analyse results as a function of sperm characteristics. In 255 couples attempting their first assisted reproductive technology (ART) attempt for male infertility (motile sperm count

Is fertility preservation feasible after the onset of puberty in adolescents with Klinefelter syndrome (KS)?Fertility preservation counseling should be an integral part of the care of XXY adolescents. Frozen ejaculated or testicular spermatozoa and even frozen immature germ cells can give them the potential to conceive their genetic progeny. However, no biological or clinical parameters were predictive of mature or immature germ cell retrieval.KS is the commonest sex chromosome disorder observed in azoospermic infertile males. Testicular sperm extraction success decreases with age and after testosterone therapy. Arguably, spermatozoa should be retrieved from KS males at the onset of puberty and before testosterone therapy to increase the chance of success.A retrospective study was performed in eight KS adolescents, aged between 15 and 17 years, who were referred for counseling about their future fertility to the center CECOS (Centre d'Etude et de Conservation des Oeufs et du Sperme humain) at Rouen University Hospital between October 2008 and December 2011.The patients were first seen with their parents and then separately. It was proposed to them that they should provide a semen sample, if this was azoospermic, two other semen samples spaced by 3 months were collected. If azoospermia was confirmed, a bilateral testicular biopsy was proposed for sperm retrieval and testicular tissue preservation. Each adolescent met the psychologist before undergoing testicular biopsy. Paraffin-embedded testicular tissue was evaluated after staining with hematoxylin-eosin and saffron and immunostaining using vimentin, anti-Müllerian hormone, androgen receptor and MAGE-A4 antibodies. Sertoli cell maturity, germ cell identification and lamina propria alteration were assessed on seminiferous tubules.KS adolescents were not deeply concerned about their future fertility and only became involved in the process of fertility preservation after at least three medical consultations. The parents agreed immediately that fertility preservation should be attempted. Seven non-mosaic XXY adolescents presented with azoospermia and one XXY/XY adolescent had oligozoospermia. Increased plasma levels of FSH and LH as well as bilateral testicular hypotrophy were observed in all patients. The XXY/XY adolescent banked four semen samples before testosterone replacement therapy. Two patients refused testicular biopsy. Five patients accepted a bilateral testicular biopsy. Spermatozoa were retrieved in one patient, elongated spermatids and spermatocytes I in a second patient.The number of patients enrolled in our study was low because the diagnosis of KS is only rarely made before or at the onset of puberty. Most XXY males are diagnosed in adulthood within the context of male infertility.Spermatozoa can be retrieved in semen sample and in testicular tissue of adolescent Klinefelter patients. Furthermore, the testis may also harbor spermatogonia and incompletely differentiated germ cells. However, the physician should discuss with the patient and his parents over a period of several months before collecting a semen sample and performing bilateral testicular biopsy. Fertility preservation might best be proposed to adolescent Klinefelter patients just after the onset of puberty when it is possible to collect a semen sample and when the patient is able to consider alternative options to achieve fatherhood and also to accept the failure of spermatozoa or immature germ cell retrieval.

Résumé Objectifs Rechercher la présence d’altérations nucléaires ou acrosomiques dans les spermatozoïdes présentant des vacuoles occupant au moins 13 % de la surface de la tête des spermatozoïdes (spermatozoïdes dits de type 3), détectées à l’aide de la technique MSOME (motile sperm organelle morphology examination). Matériel et méthodes Notre étude a été effectuée sur des recueils de sperme de 20 hommes infertiles. Une évaluation de l’acrosome par immunomarquage à l’aide d’un anticorps monoclonal antiproacrosine (Mab4D4) a été associée à une détection de la fragmentation de l’ADN par la technique TUNEL. La condensation de la chromatine spermatique a été évaluée par une coloration au bleu d’aniline, et le taux d’aneuploïdie a été déterminé par hybridation in situ en fluorescence (FISH). Une observation des échantillons spermatiques par microscopie électronique à transmission (MET) [sperme entier] et confocale [spermatozoïdes de type 3 isolés] a complété ces analyses. Les résultats ont été comparés entre des spermatozoïdes provenant du sperme entier et des spermatozoïdes de type 3 isolés. Résultats Une augmentation significative de la proportion d’acrosomes anormaux a été constatée dans les spermatozoïdes isolés de type 3 par comparaison au sperme entier (77,8 ± 2,49 % vs 70,6 ± 2,62 %). La fragmentation de l’ADN a été observée de manière comparable dans les spermatozoïdes de type 3 et le sperme entier (14,5 ± 3,45 % vs 11,5 ± 1,25%), un défaut de condensation de la chromatine ainsi qu’une augmentation significative du taux d’aneuploïdie ont été retrouvés dans les spermatozoïdes isolés par comparaison à ceux provenant du sperme entier (50,4 ± 3,10 % vs 26,5 ± 2,60 % et 7,8 ± 1,98 % vs 1,3 ± 0,18 %). Les larges vacuoles sont à localisation nucléaire exclusive (MET et microscopie confocale). Conclusion Les vacuoles seraient en relation directe avec un défaut de maturation nucléaire survenant au cours de la spermiogenèse.

Two different protocols to thaw cryopreserved human ovarian cortex have been evaluated using a histological analysis. The slower one, based on a progressive dilution of cryoprotectants, seems to maintain an optimal follicle morphology.

Resume But Comparaison des taux de testosterone et d’estradiol dans le sang du cordon spermatique et dans le sang peripherique, en fonction du resultat de l’extraction de spermatozoides testiculaires par biopsie chirurgicale chez une population de patients presentant une azoospermie non obstructive (ANO). Materiel et methodes Etude prospective, comparative, CHU promoteur, apres accord du comite d’ethique, de 30 patients presentant une ANO. Pour chaque patient, ont ete realises simultanement des dosages plasmatiques de testosterone et d’estradiol dans le sang peripherique et preleve au niveau de la veine spermatique, au moment de la biopsie testiculaire. Les analyses statistiques ont ete effectuees a l’aide du test de Mann-Whitney et du test de correlation de Spearman, par analyse de variance uni- et multivariee ( p Resultats Aucune correlation significative n’a ete retrouvee entre les taux plasmatiques peripheriques et spermatiques de testosterone et d’estradiol, quel que soit le resultat de la biopsie. Le rapport estradiol/testosterone spermatique etait significativement augmente lorsque l’extraction de spermatozoides etait negative ( p = 0,018). Conclusion L’augmentation du rapport estradiol/testosterone dans le sang spermatique en cas d’extraction negative suggere l’hypothese d’une conversion plus importante de testosterone en estradiol au niveau testiculaire. Cela pourrait temoigner d’une augmentation de l’activite aromatase en cas d’absence de cellules germinales. Des etudes supplementaires utilisant des marqueurs tissulaires de la spermatogenese devraient permettre une meilleure comprehension du role physiologique des estrogenes sur la spermatogenese et d’affiner les indications de biopsie testiculaire chez les patients presentant une ANO.

L’examen du sperme tel qu’il a ete defini par l’Organisation Mondiale de la Sante (OMS) constitue une des etapes clefs dans l’exploration du couple infertile. La morphologie du spermatozoide a ete reconnue comme le meilleur facteur predictif de la fertilite naturelle, de l’issue en insemination intra-uterine ou en fecondationin vitro classique. Les anomalies ultrastructurales des spermatozoides qui sembleraient etre les seules anomalies capables d’influencer le devenir de l’ICSI(Intracytoplasmic Sperm Injection) ne sont pas detectables aux grossissements x100 ou X200—400 utilises au cours de la procedure d’ICSI. Bartoov et al. (2002) ont mis au point un systeme capable d’etudier la morphologie fine des spermatozoides mobiles en temps reel au grossissement x6600 appele MSOME pourMagnification Motile Sperm Morphology Examination. Les anomalies detectees a ce grossissement, en dehors des anomalies morphologiques habituelles, sont des vacuoles situees sur les tetes spermatiques dont la localisation, le nombre et la taille sont variables. L’analyse en MSOME pourrait etre utilisee comme facteur predictif du pouvoir fecondant des spermatozoides et etre realisee dans l’exploration des atteintes spermatiques severes avant ICSI. L’ICSI serait classique ou precedee d’une selection fine des spermatozoides avant injection si les spermatozoides presentent un nombre eleve de vacuoles.

We report a case of laparoscopic ovarian transposition and ovarian cryopreservation in a nulliparous 25-year-old woman with rectal cancer. The authors focus on the surgical technique and the importance of preserving ovarian function.

International audience; Although electron microscopy provides a detailed analysis of ultrastructural abnormalities, this technique is not available in all laboratories. We sought to determine whether certain characteristics of the flagellum as assessed by light microscopy were related to axonemal abnormalities. Forty-one patients with an absence of outer dynein arms (type I), a lack of a central complex (type III) and an absence of peripheral doublets (type IV) were studied. Sperm morphology was scored according to David's modified classification. Flagella with an irregular thickness were classified as being of normal length, short or broken. There were correlations between missing outer dynein arms and abnormal, short or coiled flagellum. Type III patients showed the highest flagellar defects (a short (P = 0.0027) or an absent flagellum (P = 0.011)). Just over 68% of the irregular flagella were short in Type III patients, whereas this value was only 34.5% in type I and 26.4% in type IV (P = 0.002). There was a negative correlation between misassembly and spermatozoa of irregular flagella (r = -0.79; P = 0.019). It is concluded that light microscopy analysis of flagellum abnormalities may help provide a correct diagnosis, identify sperm abnormalities with fertility potentials and outcomes in assisted reproduction technologies and assess the genetic risk.

Ojective To assess the feasibility of semen cryopreservation after orchidectomy in patients with testicular tumour. Patients and methods The quality of semen samples was investigated in 36 men with testicular tumour (mean age 31.7 years, range 20–49) who were referred to our infertility centre for semen cryopreservation. For each patient, the number of straws, semen volume, number of spermatozoa, and sperm motility before and after freezing were evaluated. Results Fifteen patients (42%) banked sperm before and 21 (58%) after orchidectomy; the delay was >7 days in 19 patients (53%). The mean age, histological diagnosis and tumour stage did not differ significantly whatever the time of cryopreservation. Semen quality did not differ significantly in patients who cryopreserved sperm before or after orchidectomy and there were no significant differences in sperm values whatever the delay before preservation. Semen quality was the same in patients with seminoma or nonseminoma tumour. Conclusion These findings indicate that spermatogenesis of the contralateral testis is sufficient for successful semen cryopreservation after orchidectomy. Urologists should be encouraged to increase the awareness among oncology teams and patients about the new developments in preserving fertility for patients with cancer.

The survival of the young boy after cancer has considerably progressed in recent years due to the efficiency of chemo/radiotherapy against the tumor cells. However, this treatment causes adverse effects on healthy tissues, including fertility. Freezing testicular tissue before highly gonadotoxic treatment is a prerequisite for preserving fertility in prepubertal boys that do not produce sperm yet. But which strategy proposes to restore fertility from frozen-thawed testicular tissue? One potential solution would be to consider an in vitro maturation of spermatogonial stem cells. In this article we trace the chronological development of in vitro spermatogenesis that resulted in mouse sperm production in vitro and give an overview of new challenges for the future.

Depuis la loi du 4 juillet 2001, la vasectomie est reconnue comme methode de contraception masculine. Cette loi a autorise ce geste en l'encadrant avec des regles precises et strictes. L'AFU diffuse depuis plusieurs annees une fiche d'information-patient. L'acces a cette intervention reste cependant encore limite. Il n'existe pas de donnees publiees recentes sur la pratique de la vasectomie en France. Nous rapportons l'experience de la vasectomie et son impact medico-economique au sein d'un centre hospitalo-universitaire. Materiels et methodes : Etude de cohorte retrospective monocentrique de 45 patients ayant beneficie consecutivement d'une vasectomie contraceptive dans notre centre entre juillet 2001 et mai 2016. Pour chaque patient ont ete etudies : 1) les modalites de realisation de l'acte, 2) le respect des recommandations de MAS et de la loi de 2001, 3) les couts directs et indirects et les benefices engendres par l'intervention pour l'etablissement bases sur le libelle GHM correspondant, 4) l'efficacite du geste par l'analyse des spermogrammes de controle, 5) la satisfaction et le devenir des patients a l'aide d'un questionnaire telephonique. Resultats : L'âge moyen etait de 41,3 ans. La seconde consultation etait realisee dans 91% des cas. Le delai de reflexion n'etait pas respecte dans 24% des cas. Le consentement ecrit etait signe dans 89% des cas. La congelation preventive etait proposee dans 78% des cas. La vasectomie etait realisee en ambulatoire dans 73% des cas, sous anesthesie locale dans 6,7% des cas. Tous les patients ont eu une resection chirurgicale du deferent. Le cout moyen par patient etait de 660,63 euros (36,8% charges directes, 30,1% charges induites, 27,3% charges indirectes, 5,8% couts structure) pour un gain moyen de 524,50 euros soit une perte de 136,13 euros. Le spermogramme de controle n'etait pas effectue dans 22% des cas. Parmi les patients l'ayant fait, seulement 54,3% etaient azoospermes mais le delai de 3 mois n'etait pas respecte chez 23% d'entre eux. Aucun patient interroge n'a exprime de regret apres l'intervention. Conclusion : Dans notre experience, les recommandations issues de la loi de 2001 concernant la vasectomie n'etaient pas systematiquement suivies. Ce manque de standardisation des pratiques, potentiel reflet d'un manque d'interet, est a mettre en relief avec le surcout engendre. La revalorisation de l'acte devrait etre integree dans la reflexion d'amelioration des pratiques de sterilisation masculine.

In the management of asthenozoospermia, the spermogram-spermocytogram plays an important role during diagnosis. It is of major importance to distinguish between necrozoospermia and sperm vitality. An ultrastructural study of spermatozoa is processed in the case of primary infertility without female implication, severe, unexplained and irreversible asthenozoospermia, sperm vitality at least 50 % and normal concentration of spermatozoa. Ultrastructural flagellar abnormalities are numerous and involve most spermatozoa. ICSI provides a suitable solution for patients with sperm flagellar defects to conceive children with their own gametes but the rate of ICSI success may be influenced by the type of flagellar abnormality. Some fertilization and birth rate failures which are related to some flagellar abnormalities might occur.

Normal spermatogenesis results from a balance between process of cell proliferation, cell differentiation and apoptosis that concern somatic cells and germ cells. Dysfunction of spermatogenesis may be the result of constitutional or acquired abnormalities of spermatogonia stem cells or somatic cells. To overcome these problems, it seems necessary to implement preventive measures for germ stem cell preservation or substitute measures to replace them, the objective being to replicate in vivo or in vitro the process of spermatozoa production. This article will discuss the different experimental strategies for considering the in vivo or in vitro production of spermatozoa, outside the physiological process.

Improving our practice by a constant evaluation is essential in the field of donor semen insemination (DI). Our center examined the prognosis factors for DI success in order to standardize patient treatment options.We retrospectively analysed all couples referred for DI from January 2000 till December 2010.We analysed 551 cycles among 188 patients. Pregnancy rate by stimulation cycle was 19,8% with birth rate of 16.7%. The rate of pregnancy was improved till the fourth trial then plateau. On a patient-based analysis, success factors were age (P=0.04), previous successful DSI (P=0.02), and no previous failure of an ICSI-C (P=0.035). On a cycle-based analysis, success factors were the number of follicles greater than 15mm (P=0.04) and than 18mm (P=0.001). The percentage of 68.1 patients obtained a child by IVF-D after a failed DI.There are two predictive factors for DI success: the age of the patient and the number of mature follicles. It seems accurate to referred patients to IVF-D after four unsuccessful cycles of DSI. This recommendation may be adapted according to patient's age and hormonal evaluation.

With the motile sperm organelle morphology examination (MSOME), spermatozoa morphology may be assessed directly on motile spermatozoa at high magnification (up to 6600×). This procedure describes more precisely spermatozoa abnormalities, especially head vacuoles. However, no consensus has been established concerning normal or abnormal MSOME criteria. The aim of our study was to define MSOME vacuole criteria assessed objectively with a digital imaging system software to establish a potential relationship between conventional semen parameters. A total of 440 semen samples were obtained from males consulting in Rouen University Hospital Reproductive Biology Laboratory. Conventional semen analysis (volume, sperm concentration, progressive motility, vitality and morphology) and MSOME assessment {sperm head length, width and area as well as vacuole number, vacuole area and relative vacuole area to sperm head [RVA (%) = [vacuole area (μm(2))/head area (μm(2))] × 100)]} were performed for each semen sample. Among our 440 males, 109 presented normal conventional semen parameters and 331 abnormal ones. Sperm head vacuoles were significantly larger in abnormal semen samples (p0.0001). RVA was the most discriminative MSOME criterion between normal and abnormal semen samples according to ROC curves analysis, and was negatively correlated with poor sperm morphology (r = -0.53, p0.0001). We concluded to (i) the normal occurrence of vacuoles in sperm head whatever the normality or abnormality of semen parameters, (ii) the discriminative function of the RVA to distinguish semen samples with normal and abnormal parameters, and (iii) the strong correlation between high RVA and poor sperm morphology.

Human normal spermatozoa present a specific chromatin organization, illustrated particularly by the non-random chromosome positioning. Spermatozoa with large vacuoles, described using motile sperm organelle morphology organization (MSOME), are associated with nuclear alterations, such as abnormal chromatin condensation and aneuploidy. To question a probable association between large nuclear vacuoles and chromatin disorganization, we evaluated chromosomes X, Y and 18 topography in normal spermatozoa (NS) compared with spermatozoa with large vacuoles (SLV). After centrifugation on a gradient density system, 229 NS (spermatozoa presenting a normal nuclear shape and a vacuole area6.5% of head area) from 10 normal semen samples and 221 SLV (spermatozoa presenting a vacuole area13% of head area) from 10 semen samples with teratozoospermia were selected using MSOME. A three-colour FISH was carried out using α-satellite centromeric probes for chromosomes X, Y and 18. For each chromosome, longitudinal and spatial positioning of centromeres was analysed. Distribution of each chromosome was non-random in NS and in SLV, whatever the methodology used. Using longitudinal positioning, distribution of chromosome 18 and chromosome Y centromeres did not differ significantly between SLV and NS. On the contrary, chromosome X centromeres were more frequently positioned in the posterior region of sperm nucleus in SLV (p = 0.01). Considering spatial positioning, distributions differed significantly between SN and SLV for chromosome Y (p = 0.02) and chromosome 18 (p 10(-4) ) and marginally for chromosome X (p = 0.08). Our study concluded to a modification in chromosomes X, Y and 18 centromere topography between NS and SLV, representing a novel and supplementary evidence to argue chromatin disorganization in SLV.

To evaluate the results of the initial in vitro fertilization (IVF) procedure after intrauterine insemination (IUI) failure where half of the oocytes were inseminated as for conventional IVF and half of the oocytes where treated for intra cytoplasmic sperm injection (ICSI).From the first of January 2001 to the 31st of December 2008, 75 couples failing to conceive with ovarian stimulation and IUI were included in this retrospective study.Among the 75 couples, 53 had IVF and ICSI in sibling oocytes and 22 had just ICSI because of an insufficient oocyte cohort. No significant difference was observed in fertilization, implantation and pregnancy rates between the different groups; however, a total fertilization failure was observed more frequently when conventional IVF was used compared to ICSI (11.3% vs 3.8%). These patients with fertilization failure on conventional IVF had a lower sperm count than those who obtained embryos but sperm parameters were normal according to World Health Organisation (WHO) criteria and our study did not find any predictive factor of fertilization failure.Couples failing to conceive after controlled ovarian hyperstimulation and IUI may be proposed a mixed in vitro fertilization procedure (with conventional IVF and ICSI) in order to avoid a total fertilization failure. Moreover, this procedure will indicate the better IVF technology (conventional or ICSI) for the subsequent attempt.

BACKGROUND Sperm banking is a suitable procedure to prevent infertility after cancer therapy in male adolescents. We evaluated the feasibility of semen preservation in 156 adolescents aged between 13 and 20 years and then we assessed fertility outcome after treatment. METHODS Age, urogenital history, indications for cryopreservation, histological diagnosis and semen parameters were recorded. Fertility status after treatment was assessed by a questionnaire addressed to those patients who had utilized sperm storage. Post-treatment semen analysis was performed for 22 patients. RESULTS Cryopreservation was possible in 88.5% of cases. Azoospermia was detected in 2.6% of the patients at the time of diagnosis. Malignant disease accounted for 84% of our male adolescents. In this type of disease, semen parameters were significantly altered only among patients with metastatic malignant bone tumour. After treatment, nine patients presented azoospermia, five patients achieved pregnancy spontaneously, two achieved it after assisted reproductive technique using fresh ejaculated spermatozoa and one following sperm donation. Three failed with cryopreserved sperm. CONCLUSIONS Semen cryopreservation is possible for most adolescents and, regardless of disease type, may be a means of preserving fertility prior to gonadotoxic treatment that might impair the spermatogenesis process.

Despite technical progress in In Vitro Fertilisation (IVF) procedure, embryo implantation rate remains low. Assisted hatching has been proposed to facilitate natural embryo hatching and implantation.Our study has evaluated whether laser assisted hatching improves implantation, pregnancy and live birth rates in different cases. We studied retrospectively 143 IVF cycles concerning more than 38 years old women, 166 IVF cycles after two previous implantation failures and 180 frozen-thawed embryo transfers.Population characteristics were comparable in hatched and control groups. Implantation, pregnancy and live birth rates in women more than 38 years old were comparable with or without assisted hatching. Concerning repeated implantation failures, even if implantation, pregnancy and live birth rates were higher in assisted hatching group (FIV or ICSI), the differences were not significant. After frozen-thawed embryo transfers, implantation rate was significantly better with assisted hatching (19.14% vs 8.84% [p=0.02]).Assisted hatching improves embryo implantation rate after frozen-thawed embryo transfer.

Comparison of testosterone and oestradiol levels in spermatic cord blood and peripheral blood as a function of the results of testicular sperm extraction by surgical biopsy in a population of patients with nonobstructive azoospermia (NOA).Prospective, comparative, teaching hospital-sponsored, ethics committee-approved study of 30 patients with NOA. Plasma testosterone and oestradiol assays in peripheral blood and blood taken from the spermatic vein during testicular biopsy were performed simultaneously for each patient. Statistical analyses were performed with Mann-Whitney test and Spearman's correlation coefficient by univariate and multivariate analysis of variance (p0.05).No significant correlation was demonstrated between spermatic and peripheral blood levels of testosterone and oestradiol, regardless of the biopsy results. The spermatic oestradiol/testosterone ratio was significantly increased when testicular sperm extraction was negative (p=0.018).The increase of the oestradiol/testosterone ratio in spermatic cord blood in the case of negative testicular sperm extraction suggests the hypothesis of greater conversion of testosterone to oestradiol in the testes. This could reflect increased aromatase activity in the absence of germ cells. Further studies using tissue markers of spermatogenesis should provide a better understanding of the physiological role of oestrogens in spermatogenesis and to refine the indications for testicular biopsy in patients with NOA.

Complex chromosomal rearrangements (CCRs) are rare events in human pathology and are usually considered to induce severe reproductive impairment by disturbing the meiotic process and producing unbalanced gametes responsible for high reproductive risk. One-third of all CCRs are familial and tend to implicate fewer breakpoints and fewer chromosomes than de novo cases. CCRs are rarely transmitted through spermatogenesis and are primarily ascertained by male infertility. We report a familial balanced CCR, with seven breakpoints involving three chromosomes, which was detected prenatally in a female fetus conceived after intracytoplasmic sperm injection (ICSI) in a couple initially thought to be a carrier of a paternal reciprocal translocation involving two chromosomal breakpoints. Fluorescent in-situ hybridization (FISH) was used to elucidate the complexity of this CCR. The karyotype of the female CCR carrier was balanced and determined as 46,XX.ish t(1;4)(q42;q32)(WCP1+, D1Z5+, WCP4+, D1S3738-, D4S2930+; WCP4+, D4Z1+, WCP1+, D4S2930-, D1S3738+), ins(1;11)(q41;q23q24)(WCP1+,WCP11+, D11S2071-, MLL+; WCP11+, D11S2071+, WCP1-, MLL-), ins(4;11)(q23;q14q23)(WCP4+,WCP11+; WCP11+,WCP4-). The same balanced CCR was confirmed in her oligozoospermic father. We report, to our knowledge, the first case of ICSI performed in an infertile male with CCR, resulting in a balanced CCR carrier female with a normal clinical follow-up at 4 years of age. This particular case stresses the point of the relevance and feasibility of ICSI procedure in cases of balanced CCRs.

We present one of the very few deliveries occurring following intracytoplasmic sperm injection of thawed immotile testicular spermatozoa from a testicular biopsy of a man with bilateral congenital absence of the vas deferens. A first attempt in the 33 year old woman with fresh testicular biopsy extracted spermatozoa was unsuccessful. The supernumerary spermatozoa were cryopreserved for later use. After thawing, testicular spermatozoa were immotile. From 11 intact oocytes injected with frozen-thawed immotile testicular spermatozoa, a two pronuclear fertilization rate of 27% and a cleavage rate of 100% were obtained. A total of three embryos was transferred resulting in a singleton pregnancy and the birth of a normal female baby.

We report a case of laparoscopic ovarian transposition and ovarian cryopreservation in a nulliparous 25-year-old woman with rectal cancer. The authors focus on the surgical technique and the importance of preserving ovarian function.

An increased incidence of cancer is observed in the population of adolescents and young adults since thirty years. Major progress in cancer diagnosis and therapy is unfortunately associated to high degree of toxicity on gonad function. Cryopreservation of ovarian tissue is performed in girls and women before cancer treatment with high risk of infertility. Procedures for ejaculated or testicular extracted spermatozoa are well defined. However, for prepubertal boys or after ejaculated sperm collection failure, mature or immature testicular tissue banking should be proposed. Still, an optimal cryopreservation protocol is a prerequisite for clinical application and does not exist for the moment. Furthermore, the future applications of immature testicular tissue banking should be developed, not solely germ cell in vitro maturation but also autologous testicular tissue grafting.

Because of a possible relationship between microenvironmental disturbances and meiotic abnormalities and of a straight relationship between lower-quality semen in patient carrying a varicocele and first meiotic non-disjunction, bilateral bipolar testicular biopsies are realized according the thermic differential gradient described in varicocele. Systematic meiotic studies of multiple testicular biopsies from 65 azoospermic men with bilateral varicocele were done in a multi-centric study on microsurgical correction of bilateral varicocele with microthermic intra-operative evaluation using minimally invasive thermal microsensors (Betatherm 10K3MCD2). In the present study abnormal temperature raising, histomorphometric abnormalities (spermatocyte arrest) and meiotic abnormalities (class IIC) are strongly correlated. In the ten patients submitted to another testicular biopsy procedure six months after surgery for TESE, normal thermal differential is registered and no meiotic abnormalities recurrences are found.

Chromosome meiotic pairing during male meiosis is a major event for chromosome segregation during anaphase I and spermatogenesis normal process. Chromosome non-disjunctions responsible for aneuploidy in male gametes can be observed during the first and the second meiotic divisions. The analysis of sperm nuclei chromosome constitution is a major and indirect tool for assessing male meiotic non-disjunctions and the genesis of chromosomal abnormalities. This evaluation has been performed initially by the human sperm/hamster oocyte fusion assay and more recently by fluorescence in situ hybridisation (FISH). Therefore, male populations with increased risk of aneuploidy for their progeny could be identified before entering an in vitro fertilization procedure, and depending on the potential risk a preimplantation or prenatal genetic diagnosis could be performed. For males with constitutional chromosome abnormalities, a specific genetic counselling could also be proposed.

The aim of this study was to determine the incidence of sex chromosome aneuploidy in spermatozoa of two males with a 47,XYY karyotype and one male with a 46,XY/47,XYY constitution. Spermatozoa obtained from two oligospermic patients and one volunteer semen donor were studied by multicolour fluorescence in situ hybridization. In the XY/XYY male, the frequencies of X-bearing to Y-bearing sperm were significantly different from the 1 : 1 expected ratio. Significantly increased frequencies were found in the mosaic and non-mosaic males for 24,XX and 24,YY sperm when compared with control donors. The number of 24,XY sperm was significantly different from the controls in the XYY males, but not in the mosaic male. The incidence of disomy 18 and the rate of diploidy also increased in the three patients. However, the mosaic male had the lowest cumulative rate of disomic and diploid spermatozoa when compared with the two XYY patients. Our data suggest that: (i) chromosome abnormalities observed in spermatozoa of the two XYY oligoasthenoteratospermic (OAT) males arise through segregation errors in XY germ cells rather than normal meiosis of XYY germ cells, (ii) mosaic XYY males with normal semen parameters have a lower risk of producing offspring with a sex chromosomal abnormality than OAT males with XYY karyotype.

To assess the feasibility of semen cryopreservation after orchidectomy in patients with testicular tumour.The quality of semen samples was investigated in 36 men with testicular tumour (mean age 31.7 years, range 20-49) who were referred to our infertility centre for semen cryopreservation. For each patient, the number of straws, semen volume, number of spermatozoa, and sperm motility before and after freezing were evaluated.Fifteen patients (42%) banked sperm before and 21 (58%) after orchidectomy; the delay was7 days in 19 patients (53%). The mean age, histological diagnosis and tumour stage did not differ significantly whatever the time of cryopreservation. Semen quality did not differ significantly in patients who cryopreserved sperm before or after orchidectomy and there were no significant differences in sperm values whatever the delay before preservation. Semen quality was the same in patients with seminoma or nonseminoma tumour.These findings indicate that spermatogenesis of the contralateral testis is sufficient for successful semen cryopreservation after orchidectomy. Urologists should be encouraged to increase the awareness among oncology teams and patients about the new developments in preserving fertility for patients with cancer.

Fluorescence in situ hybridization (FISH) using whole-chromosome paint probes was performed to evaluate disomy and diploidy frequency for chromosomes 1, 18, 19, and 22 in human sperm nuclei.Ten subjects of proven fertility and normal spermatic parameters were included in the study. A dual-color FISH method was carried out.A total of 157,896 spermatozoa was scored. The mean frequencies of disomic sperm for chromosomes 1, 18, 19, and 22 were 0.22% (range, 0.19 to 0.28%), 0.24% (range, 0.14 to 0.37%), 0.22% (range, 0.17 to 0.30%), and 0.25% (range, 0.21 to 0.29%), respectively. The mean frequency of diploidy was 0.14% (range, 0.09 to 0.18%). No interindividual and interchromosomal variations in the aneuploidy frequency were observed between the different subjects.FISH with whole-chromosome paint probes provides a novel and efficient approach for disomy assessment in human sperm nuclei.