Cell-free DNA in human follicular fluid as a biomarker of embryo quality

International audienceStudy question: Could cell-free DNA (cfDNA) quantification in individual human follicular fluid (FF) samples become a new non-invasive predictive biomarker for in vitro fertilization (IVF) outcomes?Summary answer: CfDNA level in human follicular fluid samples was significantly correlated with embryo quality and could be used as an innovative non-invasive biomarker to improve IVF outcomes.What is known already: CfDNA fragments, resulting from apoptotic or necrotic events, are present in the bloodstream and their quantification is already used as a biomarker for gynaecological and pregnancy disorders. Follicular fluid is important for oocyte development and contains plasma components and factors secreted by granulosa cells during folliculogenesis. CfDNA presence in follicular fluid and its potential use as an IVF outcome biomarker have never been investigated.Study design, size, duration: One hundred individual follicular fluid samples were collected from 43 female patients undergoing conventional IVF (n = 26) or ICSI (n = 17). CfDNA level was quantified in each individual follicular fluid sample.Participants/materials, setting, methods: At oocyte collection day, follicles were aspirated individually. Only blood-free follicular fluid samples were included in the study. Follicle size was calculated based on the follicular fluid volume. Each corresponding cumulus-oocyte complex was isolated for IVF or ICSI procedures. Follicular fluid cfDNA was measured by quantitative PCR with ALU-specific primers.Main results and the role of chance: Human follicular fluid samples from individual follicles contain measurable amounts of cfDNA (mean ± SD, 1.62 ± 2.08 ng/µl). CfDNA level was significantly higher in small follicles (8-12 mm in diameter) than in large ones (>18 mm) (mean ± SD, 2.54 ± 0.78 ng/µl versus 0.71 ± 0.44 ng/µl, respectively, P = 0.007). Moreover, cfDNA concentration was significantly and negatively correlated with follicle size (r = -0.34; P = 0.003). A weak significant negative correlation between DNA integrity and 17β-estradiol level in follicular fluid samples at oocyte collection day was observed (r = -0.26; P = 0.008). CfDNA level in follicular fluid samples corresponding to top quality embryos was significantly lower than in follicular fluid samples related to poor quality embryos (P = 0.022). Similarly, cfDNA level was also significantly lower in follicular fluid samples related to embryos with low fragmentation rate (≤25%) than with high fragmentation rate (>25%) (P = 0.02).Limitations, reasons for caution: A larger study should be conducted in order to establish the predictive value of cfDNA level for embryo quality and to investigate whether follicular fluid cfDNA levels are correlated with embryo implantation rates and pregnancy outcomes. Moreover, the role of follicular fluid cfDNA on embryo quality should be studied to determine whether high cfDNA concentration in follicular fluid is only a consequence or also a cause of follicular dysfunction.Wider implications of the findings: CfDNA evaluation in individual follicular fluid samples might represent an innovative biomarker of embryo quality to use as a supplemental tool to predict embryo quality during IVF.Study funding/competing interests: This study was partially supported by the University Hospital of Montpellier and Ferring Pharmaceuticals. The authors of the study have no competing interests to report

Embryons vitrifiés, embryons frais : comparaison des poids de naissance

International audienceIt is already known that children born after slow frozen embryo replacement have a significantly higher birth weight compared to children born after fresh embryo transfer. Similar data have been reported related to frozen embryo transfer using an open vitrification system. However, few data relative to birth weight using a complete embryo closed vitrification system has been reported. The purpose of this study was to know if frozen embryo in closed vitrification system is associated with a higher birth weight compared to fresh embryo replacement.Design: This was a monocentric retrospective cohort study, 371 children were issued from fresh embryo replacement and 127 from vitrified embryo transfer.Materials and methods: All singletons born after fresh or vitrified embryo transfer between January 2011 and April 2015 were included. Births from the vitrified or fresh transfers of egg or sperm donation, and preimplantation genetic diagnosis were excluded. In addition, pregnancies with more than one gestational sac at the first ultrasound were excluded. An analysis of covariance (ANCOVA) was used for multivariate analysis.Results: Mean birth weight was 205g higher in the frozen embryo compared with fresh embryos transfer groups (3368g vs. 3163g respectively, P<0.001). This difference remained after multivariate analysis adjusted on confounding factors such as gestational age, maternal age, maternal body mass index (BMI), tobacco exposure, number of embryo transferred and birth order (P<0.001)..Conclusions: Embryo frozen in closed vitrification system is associated with a higher birth weight compared to fresh embryo replacement. Our findings are consistent with the previous studies related to slow freezing and open vitrification systems data. The effects of controlled ovarian stimulation (COS), ex vivo culture conditions and cryopreservation systems on birth weight of children born should be further explored.Les enfants nés après un transfert d’embryon congelé par congélation lente ou vitrifié en système ouvert ont un poids de naissance plus élevé que les enfants nés suite à un transfert d’embryon frais. Cependant, il existe peu de données sur le poids de naissance et la vitrification en système fermé. Cette technique de vitrification est la plus utilisée en France. L’objectif de cette étude est de savoir si la vitrification embryonnaire en système fermé était associée à des poids de naissance plus élevés que les transferts d’embryons frais.Méthodes :Il s’agit d’une étude unicentrique de cohorte rétrospective. Tous les singletons nés suite à un transfert d’embryon frais ou vitrifié en dehors d’un don de gamète entre janvier 2011 et avril 2015 ont été inclus. Les embryons ayant bénéficié d’un diagnostic pré-implantatoire ou dont la grossesse était initialement gémellaire ont été exclus. Une analyse de covariance (ANCOVA) a été utilisée pour l’analyse multivariée.Résultats :Nous avons étudié 371 singletons nés après transfert d’embryons frais et 127 singletons nés après transfert d’embryons vitrifiés en système fermé. Le poids de naissance des enfants nés après replacement d’embryons vitirifés était significativement plus élevé de 205 g en comparaison avec les enfants nés après replacements d’embryons frais (3368 g vs 3163 g respectivement, p < 0,001). Cette différence était toujours significative après analyse multivariée incluant les facteurs confondants que sont l’âge maternel, l’IMC maternel, la parité, l’exposition tabagique et le nombre d’embryons transférés (p < 0,001).Conclusion :Les embryons vitrifiés en système fermé sont associés à un poids de naissance plus élevé que les embryons frais. Ces données sont cohérentes avec les données de la congélation lente et la vitrification en système ouvert. Les effets de l’hyperstimulation ovarienne contrôlée, les conditions de culture ex vivo et les effets de la cryopréservation sur le poids de naissance nécessitent d’être étudiés plus largement

La réceptivité endométriale en aide médicale à la procréation : Une piste à ne pas oublier devant un échec d’implantation

International audienceInfertility affects between 8 and 12% of reproductive-age couples worldwide. Despite improvements in assisted reproductive techniques (ART), live birth rates are still limited. In clinical practice, imaging and microscopy are currently widely used, but their diagnostic effectiveness remains limited. In research, the emergence of innovative techniques named OMICS would improve the identification of the implantation window, while progressing in the understanding of the pathophysiological mechanisms involved in embryo implantation failures. To date, transcriptomic analysis seems to be the most promising approach in clinical research. The objective of this review is to present the results obtained with the different approaches available in clinical practice and in research to assess endometrial receptivity in patients undergoing ART.L’infertilité touche entre 8 et 12 % des couples en âge de procréer à travers le monde. Malgré l’amélioration des techniques d’aide médicale à la procréation (AMP), les taux de naissances vivantes restent toujours limités. En pratique clinique, l’imagerie et la microscopie sont actuellement largement utilisées, mais leur efficacité diagnostique reste limitée. En recherche, l’émergence des techniques innovantes dites OMICS permettrait d’améliorer l’identification de la fenêtre implantatoire, tout en progressant dans la compréhension des mécanismes physiopathologiques impliqués dans les échecs d’implantation embryonnaire. À ce jour, l’approche transcriptomique semble être la piste la plus prometteuse en recherche clinique. L’objectif de cette revue est de faire un état des lieux des résultats obtenus à ce jour avec les différentes approches disponibles en pratique clinique et en recherche pour évaluer la réceptivité endométriale chez les patientes prises en charge en AMP

Poids de naissance et transfert d’embryon congelé : état de l’art

International audienceThe aim of this study was to update our acknowledgment if there is a link between assisted embryo cryopreservation and epigenetics in human? Animal studies have demonstrated epigenetics consequence and especially imprinting disorders due to in vitro culture. In human, it is important to note that after frozen embryo transfer birth weight is significantly increased by 81 to 250g. But these studies cannot identify the reasons of such difference. This review strongly suggests that embryo cryopreservation is responsible for birth weight variations but mechanisms not yet elucidated. Epigenetics is probably one of these but to date, none study is able to prove it. We have to be attentive on a possible link between assisted reproductive technology (ART) and epigenetics reprogrammation

Lésions périnéales sév\u27res liées á une extraction foetale par spatules. Quels facteurs de risques ?

Objective To assess risk factors for anal sphincter injury during operative vaginal delivery using spatulas. Patients and methods A monocentric retrospective study of all assisted vaginal deliveries using Thierry and Teissier\u27s spatulas between January 1st, 2008 and December 31st, 2009 in a teaching level III maternity. We studied risk factors such as primiparity, gestational age, maternal age, previous perineal laceration, level and type of presentation, type of expulsion, unsuccessful extraction and successive use of tools, episiotomy, type of anaesthesia and birth weight. Results There were 346 perineal tears (60.5%); among them, 175 (31%) were type 1, 131 (23%) type 2, 35 (6.1%) type 3 and five (0.9%) type 4. There were 235 episiotomy (41.1%). There was no statistically significant difference between all the supposed risk factors and the severe perineal tears. Conclusion There are no relationship between third and fourth degree perineal lesions during spatula\u27s delivery and supposed risk factors of anal sphincter injury. Only statistical tendances between first vaginal delivery and anal sphincter injury and between occipitosacral delivery and anal sphincter injury were found. We need further randomized studies comparing assisted births using spatulas, forceps and vacuum extractors to better assess perineal tears risk factors

Dichorio-Triamniotic Triplet Pregnancy after Day 3 Single Embryo Transfer

International audienceA 31 year-old woman had a single embryo transfer on day 3 after assisted hatching (AH) and intracytoplasmic sperm injection (ICSI) treatment. Ultrasound examination performed 6 weeks after oocyte retrieval revealed a triplet pregnancy combining monochorionic diamniotic twins and a singleton. If zygote splitting resulting in monochorionic triamniotic triplets following IVF has already been described, this case is about an incredibly rare phenomenon after single embryo transfer. Naturally, a concurrent spontaneous conception cannot be excluded. To our knowledge, this is the first time a dichorionic triplet pregnancy after single embryo transfer is reported

Programme de préservation de la fertilité féminine au CHRU de Montpellier 2ans après

International audienceOBJECTIVE:Female fertility preservation in the context of cancer management is crucial for patient's health care. The aim of this study was to evaluate the oncofertility practice at our university hospital of Montpellier since 2011.PATIENTS AND METHODS:The evaluation of management of young patients referred to Montpellier University Hospital from September 2011 to September 2013 for oncofertility counselling before cancer treatment.RESULTS:Seventy-one patients were referred to a specialized oncofertility center. Forty-two patients (59.1%) were included in the oncofertility program. Twenty-two patients (31%) were proposed for oocyte vitrification after COS protocol, eight patients (11.3%) for ovarian tissue cryoconservation, seven patients (9.9%) for GnRH injections, three patients (4.2%) ovarian transposition and two patients (2.8%) for embryo cryopreservation. Among the 42 indications of fertility preservation, only 18 will have finally taken place.CONCLUSION:Oncofertility counselling for young patients should now be part of the cancer management. It involves multidisciplinary teams. Further information of both oncologists and patients is needed to improve this new approach in the field of cancer treatments