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Objectives: To provide clinical practice guidelines about fertility preservation (FP) for women with benign gynecologic disease (BGD) developed by a modified Delphi consensus process for oocyte vitrification in women with benign gynecologic disease., Methods: A steering committee composed of 14 healthcare professionals and a patient representative with lived experience of endometriosis identified 42 potential practices related to FP for BGD. Then 114 key stakeholders including various healthcare professionals (n=108) and patient representatives (n=6) were asked to participate in a modified Delphi process via two online survey rounds from February to September 2020 and a final meeting. Due to the COVID-19 pandemic, this final meeting to reach consensus was held as a videoconference in November 2020., Results: Survey response of stakeholders was 75 % (86/114) for round 1 and 87 % (75/86) for round 2. Consensus was reached for the recommendations for 28 items, that have been distributed into five general categories: (i) Information to provide to women of reproductive age with a BGD, (ii) Technical aspects of FP for BGD, (iii) Indications for FP in endometriosis, (iv) Indications for FP for non-endometriosis BGD, (v) Indications for FP after a fortuitous diagnosis of an idiopathic diminished ovarian reserve., Conclusion: These guidelines provide some practice advice to help health professionals better inform women about the possibilities of cryopreserving their oocytes prior to the management of a BGD that may affect their ovarian reserve and fertility., Study Funding/competing Interest(s): The CNGOF (Collège National des Gynécologues Obstétriciens Français) funded the implementation of the Delphi process., (Copyright © 2022 Elsevier Masson SAS. All rights reserved.)

Objectives: Polycystic ovarian syndrome (PCOS) brings complications in the management of the assisted reproductive technology (ART) because of an oocyte quality probably impaired due to modifications of intra- and extra-ovarian factors. Our study aimed to investigate the extended culture in PCOS patients and its influence on the cumulative live birth rates., Methods: Fifty-nine PCOS patients (as defined by the Rotterdam criteria) and 114 normo-ovulatory patients (i.e. with tubal, male or idiopathic infertility, regular cycles and AMH>2ng/mL) aged<37years old who underwent a 1st or 2nd ART attempt with extended culture to day 6 were included from October 2015 to December 2017. The blastulation and cumulative live birth rates were compared between the two groups., Results: The PCOS and control patients were 32.22 and 32.91years old respectively (P=0.05). The median number of oocytes retrieved was significantly higher in the PCOS group and the median oocyte maturity rate significantly lower compared with controls. The blastulation rates were similar between the PCOS and the control groups, respectively 57.8% vs. 58.6%, P=0.88. Because of the risks of hyperstimulation syndrome, a freeze all strategy was achieved for 38.9% of PCOS patients vs. 14.0% of the control patients (P<0.01). The cumulative live birth rates were not statistically different: 31.7% in the PCOS group vs. 37.2% in the control group, P=0.50., Conclusions: PCOS was not observed to affect the extended culture nor the cumulative live birth rates in comparison to normo-ovulatory patients, supporting the blastocyst transfer strategy as a suitable option to PCOS patients., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

For more than a decade, the existence of ovarian stem cells that can contribute to neo-oogenesis in the adult ovary is reported by some teams, challenging the dogma according to which mammalian females are born with a fixed and non-renewed germinal cell pool. The presence of germinal stem cells with mitotic activity suggests the possibility of potential postnatal oogenesis. These cells have both germ-line and stem cell markers in culture. They have been isolated using different strategies and their ability to differentiate into oocytes has been demonstrated since after reintroduction in an ovarian somatic environment, these cells generate follicles capable of producing healthy offspring in rodents. However, many scientists remain skeptical and question the reliability of the methods used. Despite that there is no consensus on the origin of these ovarian stem cells, private companies are now proposing to use their stem cell potential to treat human infertility., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

The concept of Air Quality often refers to particulate and microbiological contamination of ambiant air. European Directive 2006/86/CE encompass the IVF process and specify a class A air quality for manipulation of tissue and cells, in a class D environment (A over D rule). Recognizing the paramount importance of ensuring the highest microbiological and particulate safety in the IVF laboratory, it is equally important to take into account practicability issues and the financial burden of these recommendations, as well as the utter need to protect gametes and embryo viability during their IVF journey. The usefulness of such stringent recommendations may also be questionned given the absence of published cases of airborne contaminations and related patients infections after embryo transfer. The European directive stems from pharmaceutical standards and were not specifically designed for human IVF. Gametes and embryos are indeed extremely sensitive to physical and chemical stress and require strict temperature, osmolarity and pH control, as well as an absence of chemical contamination during manipulation and culture. These conditions are hardly obtained when using laminar flow hoods. Following concerns raised by many experts in the field, exceptions to the A over D rule were added in the 2006/86/CE Directives. This narrative review discusses all these aspects in a critical way and compare scientific and legal requirements applying to IVF practices in different regions of the world., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

The frequency of polycystic ovary syndrome (PCOS) and the consequent fertility disorders cause many difficulties in the management of the assisted reproductive technics. Some studies are focused on different additional treatments, stimulation protocols or techniques that could optimize the in vitro fertilization cycles. The quality of the oocytes and embryos of these patients is also an outstanding issue. They remain difficult to actually evaluate during management, and none of the few published studies on this subject demonstrated any inferiority, compared to control patients. However, many differences have been highlighted, studying intra- and extra-ovarian factors. The advent of new genetic techniques could allow a better understanding of the pathophysiological mechanisms of the syndrome, as well as refining the evaluation of oocytes and embryos, in order to better predict the results of in vitro fertilization attempts. Pregnancy and birth rates, however, appear to be comparable to those of the general population., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Objectives: To investigate prospectively the pattern of the follicular growth and to characterize the COH outcome in terms of oocyte number and maturity in patients with voluminous recurrent benign ovarian tumors with a high surgical risk of significant reduction of the ovarian follicular content., Methods: The inclusion criteria were: age between 18 and 36, presence of at least one benign ovarian tumor (≥ 5cm) with high risk of recurrence. The fertility preservation cycle was performed at least 3 months after the cyst surgery. The controlled ovarian stimulation was performed after the ovarian reserve was assessed (AMH measurement and sonographic antral follicle count). Triggering was performed by hCG when at least 3 follicles reached 18mm of diameter. Metaphase II oocytes were cryopreserved by the vitrification technique., Results: Twenty-four women with dermoid, endometrioma or seromucinous cysts were included from January 2015 to July 2016. All of them had previous ovarian surgery. Mean AMH levels were 15.3pmol/L. The mean number of total oocytes retrieved was 7±5. The mean number of metaphase II oocytes was 4.4±4. The incidence of low ovarian response was 38%. Among the patients, 86% had less than 8 metaphase II oocytes vitrified. Seven patients asked for a second cycle in order to have more oocytes., Conclusion: We demonstrated the feasibility of the systematic proposal of fertility preservation by oocyte cryopreservation in this group of young patients with recurrent ovarian benign tumors. Taking into account history of previous surgery and high incidence of low ovarian reserve, the ovarian response under stimulation was frequently poor with, as consequence, low retrieved oocyte number per cycle. An oocyte accumulation strategy is then proposed to enhance further pregnancy chances., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

The oocyte grows within a follicle composed of layers of somatic cells. It undergoes with the cumulus cells that form the innermost layer a dialogue that is critical for its maturation. Based on the assumption that the transcriptome of the cumulus cells reflects the physiology of the oocyte, it may prove a useful non-invasive tool in embryo selection to improve assisted reproduction outcomes. During the past decade, various studies have been conducted with the objective of identifying cumulus biomarker genes as prognosis tools for oocyte quality and competence. Remarkably no common biomarkers stand out among all these studies. In this review we perform a critical analysis of the literature in order to reveal some of the parameters that may account for these discrepancies, such as patients' inclusion criteria (maternal age, stimulation protocols), day of embryo transfer (day 3 or 5), outcome criteria (oocyte potential, embryo competence, pregnancy). Moreover there is a lack of standardization in the experimental designs used for RNA extraction and gene expression assessment (microarrays, RT-qPCR) and for the statistical analyses. In conclusion, critical analyses such as the present one are indispensable to pave the way for future searches of predictive biomarkers of pregnancy., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Objectives: Oocyte vitrification using an open device is thought to be a source of microbiological and chemical contaminations that can be avoided using a closed device. The principal purpose of this study was to compare the two vitrification protocols: closed and open system. The secondary aim was to study the effects of the storage in the vapor phase of nitrogen (VPN) on oocytes vitrified using an open system and to compare it to those of a storage in liquid nitrogen (LN)., Methods: Forty-four patients have been included in our study between November 2014 and May 2015. Two hundred and fourteen oocytes have been vitrified at germinal vesicle (GV), metaphase I (0PB) and metaphase II (1PB) stages. We vitrified 96 oocytes (59 GV/37 0PB) using a closed vitrification device and 118 oocytes (57 GV/31 0PB/30 1PB) using an open device. The vitrified oocytes were then stored either in LN or in VPN. The main outcome measures were the survival rate after warming (SR), meiosis resumption rate (MRR) and maturation rate (MR)., Results: The global post-thaw SR was significantly higher for oocytes vitrified using an open system (93.2%) compared to those vitrified using a closed one (64.5%; P<0.001). On the contrary, there was no significant difference in terms of global MRR and MR (82.1% vs. 87.5% and 60.7% vs. 61.2% using closed and open system respectively). The SR, MRR and the MR were not significantly different when vitrified oocytes were stored in VPN or LN (91.6, 83.8, 64.5% vs. 93.9, 89.8, 59.1% respectively)., Conclusion: Taking into account the limits of our protocol, the open vitrification system remains the more efficient system. The use of sterile liquid nitrogen for oocyte vitrification and the subsequent storage in vapor phase of nitrogen could minimize the hypothetical risks of biological and chemical contaminations., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Objective: The aim of this study was to compare embryo development cultured in two single-step media commercially available: Fert/Sage One Step® (Origio) and Continuous Single Culture® (CSC) (Irvine Scientific)., Methods: A prospective auto-controlled study of sibling oocytes from women undergoing conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) was performed in our center from February to June 2015. After fertilization, for every patient, half of oocytes were cultured in the single-step Fert/Sage One Step® (serie SAGE) and the other half in the single-step CSC®(serie CSC). Fertilization and embryo morphology rates were assessed by day 1 to day 5-6 if needed. Embryo presenting<20% of fragmentation and 4 cells at day 2, 8 cells at day 3 were qualified as "top quality". Embryo with<20% of fragmentation and 3-5 cells at day 2, 6-10 cells at day 3 were qualified as "good quality". Blastocyst B3, B4, B5 with A or B inner cell mass and A or B trophectoderm were qualified as "good quality". Transferred or frozen embryos were qualified as useful embryos., Results: Sixty-two attempts of IVF and 133 of ICSI were analyzed, corresponding to 2059 inseminated or micro-injected oocytes. Fertilization rate were not different between the 2 series, respectively SAGE vs CSC (IVF: 73.4% vs 68.3% [P=0.49]; ICSI: 58.9% vs 63.8% [P=0.12]). No difference was found for embryo morphology, respectively SAGE vs CSC, at day 2 (top quality embryo at day 2 IVF: 34.4% vs 33% [P=0.98]; ICSI: 42.4% vs 44.9% [P=0.37]; and good quality embryo at day 2 IVF: 44% vs 50.2% [P=0.07]; ICSI: 64% vs 71% [P=0.35]); no difference at day 3 (top quality embryo at day 3 IVF: 19.4% vs 21.3% [P=0.61]; ICSI: 28.7% vs 27.4% [P=0.54]; and good quality embryo at day 3 IVF: 40.4% vs 50.2% [P=0.91]; ICSI: 51% vs 47.6% [P=0.47]). Blastocyst development rate were not different, respectively SAGE vs CSC (IVF: 39.9% vs 41.5% [P=0.63] with 42.9% vs 42.2% of good quality blastocyst [P=0.70]; ICSI: 41.1% vs 37.8% [P=0.18] with 32.9% vs 40.8% of good quality blastocyst [P=0.13]). No difference was found in the useful embryo rate in the 2 series SAGE vs CSC (IVF: 52.8% vs 55.2% [P=0.83]; ICSI: 62.4% vs 61.7% [P=0.70])., Conclusion: Embryo development and rate of useful embryos, transferred or frozen, were not different according to the embryo culture in single-step media Fert/Sage One Step® vs single-step Continuous Single Culture®., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

In severe male infertility, in vitro fertilization (IVF) with intra-cytoplasmic sperm injection (ICSI) represents the sole available therapeutic option. However this technique is not always successful in promoting fertilization, as some couples completely and repeatedly fail to obtain any embryo. In many cases, this failure can be attributed to a defective rise in intracellular calcium, which is required to achieve oocyte activation. Over the last twenty years, several laboratories dedicated to assisted reproduction technologies have been using a calcium ionophore to assist oocyte activation. The aim of this review is to give an overview of the advances and consequences associated with this new technique referred to as assisted oocyte activation., (© 2016 médecine/sciences – Inserm.)

Sperm and embryos cryopreservation is a commonly applied technique for several years. Recently authorized in France, vitrification tends to replace gradually the conventional technique of slow freezing, so upsetting the practices in the management of patients. It allows from now on the cryopreservation of oocytes and opens new perspectives in egg donation either still in fertility preservation. This review thus attempted to examine the contribution of vitrification in the freezing of oocytes and human embryos at various stages of development. If obviously vitrification appears as the current method of choice for the cryopreservation of oocytes as well as blastocysts, the results are less cut as regards embryos to early stages. No increase in adverse obstetric and perinatal outcomes in children conceived from vitrified oocytes or embryos is noted in the literature., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

Oocyte vitrification has been authorized in France after the modification of French bioethics law in July 2011. This evolution will bring a dramatic change in patients' management since, from 2011, infertile couples, oocyte donation and fertility preservation programs will benefit this technique in France. We have introduced oocyte vitrification in our ART laboratory through a validation of the method using Evidence-Based Medicine model: open system Cryotop, Ethylène-glycol 15% and DMSO 15%. Based on our 1-year experience, oocyte vitrification upgrades our daily practice while also minimizing embryo cryoconservation as recommended by the law., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Objective: Our objective was to study the relationships between the characteristics of the movement of sperm and their attachment to the zona pellucida., Materials and Methods: Study related to 47 patients candidates with an IVF protocol. The puncture of the ovocytes was carried out under general anesthesia under echography control after stimulated cycles using association is the citrate of clomifene and the HMG or the busereline and the HMG. Study of the mobility of the spermatozoa was carried out thanks to a videomicrography computer-assisted before and after treatment of sperm in Percoll. The setting in contact of the spermatozoa with the ovocytes having given failures of in vitro fertilization and beforehand preserved in a saline solution was carried out during 90 minutes with 37 °C, air-CO2 with 5%. After incubation, the spermatozoa fixed at the zona pellucida, are counted thanks to a meter with hand., Results: The study of the test of fixing of the spermatozoa at the zona pellucida showed that the interaction spermatozoa/ovocytes is correlated positively with the parameters of the mobility analyzed by videomicrography computer-assisted: Vcl, Vsl and the ALH., Conclusion: The performance of the test requires a preliminary epididymal maturation of the spermatozoa in particular the ontogenesis of the spermatic sites of recognition for the zona pellucida. A deterioration of this stage of maturation led to failures of recognition and fixing at the zona pellucida. The test can predict the outcome of IVF and towards the good technique of medical assistance to procreation (AMP)., (Copyright © 2012. Published by Elsevier Masson SAS.)

This review analyzes the literature concerning oocyte and embryo quality, in case of in vitro fertilization (IVF) for women with polycystic ovary syndrome (PCOS). Alterations in oocyte quality, and consequently in embryo quality, may be due to endocrine and intra-ovarian paracrine changes. However, most of publications find similar biological and clinical results after IVF, with or without microinjection, for women with PCOS compared to those obtained in control populations. Subgroups of more pejorative outcome probably exist within PCOS population. Finally, obesity, which is frequent in PCOS, is clearly deleterious, and multidisciplinary care, including lifestyle modifications, is then needed., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Worldwide statistics agree that at least one out of six couples has fertility problems. If the male gamete is the origin of this problem, it is generally admitted that the oxidative stress is involved. Modern life has obviously increased fertility problems through pesticides, xenoestrogenes, endocrine disrupting chemicals involved in plastic technology such as polychlorinated bisphenyls, bisphenol A, phthalates and alkylphenols… and other cosmetic additives. An important part of these compounds increases oxidative stress, at least in part. Oxidative stress is more than probably at the origin or recurrent increasing pathologies such as endometriosis. If the oocyte is theoretically able to repair oxidative stress linked decays such as DNA fragmentation and oxidation of bases, its capacity is finite and decreasing with age. In order to decrease DNA repair charge, reducing or even avoiding the generation of DNA damages related to reactive oxygen species through consumption of antioxidants compounds is often tempting: however Reasons will be provided to break from current treatments given haphazardly in the population in the age of reproduction, as well as the potential risks of over-exposure. Furthermore recommended treatments, in relation with the new concepts in oxidative stress, will be specified., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Oocyte vitrification is a new technique in cryobiology that will lead to a number of improvements in assisted reproduction practices, oocyte donation and the preservation of female fertility. Professionals already versed in the techniques of micromanipulation will be able to master the new procedures, which should not be delegated to unqualified staff. When adopted by clinical units, oocyte vitrification will require changes in laboratory and administrative organization. The technique will also modify the ethical outlines of reproductive biology. France today is running behind in the application of this major development in cryobiology. The reasons are many and have to do with a long waiting period for authorization from national health authorities, a lack of material and human resources and a foreseeable shake-up in the nationally established egg donation program. However, recently a new law of bioethics has recognized the breakthrough that this new technique represents by allowing couples covered by the French national health care program for Assisted Reproductive Technologies (ART) to choose oocyte vitrification as an option., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

The purpose of this review is to present the strategies for fertility preservation that are currently proposed to patients at risk from premature ovarian failure (POF) due to chemotherapy as well as some future solutions that could allow patients already affected by an IOP to recover their fertility. Today, cryo-preservation of pieces of ovarian cortex followed by orthotopic transplantation as well as in vitro maturation of oocytes (IVM) allow preservation of fertility in patients being to undergo an antitumor treatment. However, although promising results have been obtained, the pregnancy rate remains very low. Improvement of these techniques, but also stimulation of resting follicle activation, in vitro folliculogenesis, and culture of putative oogonia that may be present in the adult ovary, are all future opportunities for patients suffering from an IOP, and are currently the subject of intensive researches., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Oocyte donation is offered to patients with premature ovarian failure to achieve pregnancy when no other assisted reproductive technology is possible. Some clinical and biological factors have been identified for influencing the outcome of oocyte donation cycles. Embryo implantation depends on embryo quality, method for the embryo transfer, and endometrial differentiation. In our center, the oocyte recipients receive for the endometrial preparatory cycle the same treatment that for the patients undergoing frozen embryos transfers, with good clinical pregnancy rates, about 35% per transfer. Estrogen and progesterone supplementation with GnRH agonist down regulation are used in synchronized protocols or for frozen embryos transfers. The synchronization between recipient's endometrium and donor's ovarian stimulation is very restrictive. Nowadays, the oocytes vitrification lithens the oocyte donation process: the endometrial preparation has a limited duration and is well controlled, and embryos that are transferred are all fresh embryos., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Ovarian reserve assessment has become critical when counselling infertile couples. This evaluation should be conducted in all women over 35 years, or earlier in case of familial history of premature ovarian failure, gonadotoxic treatment of short menstrual cycles. The first step for ovarian reserve assessment is to measure basal hormonal status, i.e. day 3 FSH and estradiol. If necessary, antral follicle count performed by a skilled operator and/or serum AMH level can give some relevant additional information. The historical concept of 2-years waiting does not make sense any more regarding the current delay in childbearing. Therefore, the infertility workup, and particularly ovarian reserve assessment, should be initiated after 6 months of infertility in all women over 35 years of age.

Although female cancer incidence may be on rise, antineoplastic regimens have become more successful. As a result, an increasing number of women with cancer survive to endure the long-term consequences of chemotherapy. One of the most important long-term consequences of cancers treatments in young female is premature ovarian failure and infertility. Because of the increasing survival rates, many of these young women are seeking methods to preserve their fertility. Currently, embryo/oocytes cryoporeservation obtained after ovarian stimulation appears to provide the best fertility preservation option. However, patients may not have sufficient time to undergo ovarian stimulation prior to chemotherapy and/or the hormones used in ovarian stimulation are contra-indicated for estrogen-dependant tumors. In vitro maturation of oocytes (IVM) has been suggested to avoid ovarian stimulation and time requirement in patients with cancer, and can be combined with ovarian tissue cryobanking. In this review, we will discuss the position of IVM in the strategy of fertility preservation in young women., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)

The production of fertilizable ova is the consequence of multiple events that start as soon as ovarian development and culminate at the time of ovulation. Throughout their development, germ cells are associated with companion somatic cells, which ensure germ cell survival, growth and maturation. Data obtained in vitro and in vivo on several animal models of germ cell depletion have led to uncover the many roles of germ cells on both ovarian development and folliculogenesis. During ovarian development, germ cells become progressively enclosed within epithelial structures called "ovigerous cords" constituted by pregranulosa cells, lined by a basement membrane. At the end of ovarian development, ovigerous cords fragment into primordial follicles, which are epithelial units constituted by an oocyte surrounded by a single layer of granulosa cells. Germ cells are necessary for the fragmentation of ovigerous cords into follicles, since in their absence, no follicle will form. Germ cells also ensure the differentiation of the ovarian somatic lineage, and they may inhibit the testis-differentiating pathway by preventing the conversion of pregranulosa cells into Sertoli cells, their counterpart in the testis. Regularly, primordial follicles are recruited into the growing follicle pool and initiate their growth. They develop through primary, preantral, antral and preovulatory stages before being ovulated. Interestingly, the action of the oocyte on companion somatic cells tightly depends on the follicular stage. In primordial follicles, the oocyte prevents the transdifferentiation of granulosa cells into cells resembling Sertoli cells. By contrast, as soon as the follicle enters growth, the oocyte regulates the functional differentiation of granulosa cells and at the latest stages, it prevents their premature maturation into luteal cells. Overall, these data demonstrate that the female germ cell act on companion somatic cells to regulate ovarian development and folliculogenesis, thereby actively supporting its own maturation.

The use of high levels of cryoprotectants (CPs) in solutions applied to vitrify oocytes or embryos is an argument to still prefer slow freezing procedure. Is it a justified argument? Out of three studies using mice zygotes we may assume that (i) the intracellular concentration of CPs is far lower than the one in the vitrification solutions, (ii) the intracellular concentration of CPs in the vitrified zygote is in contrary to the common beliefs even lower than the one observed after a slow freezing procedure, (iii) survival after slow freezing reflects the presence of an intracellular vitrified state in these cells., (Copyright © 2010. Published by Elsevier SAS.)

Assisted Reproductive Techniques (ART) separating oocytes in sibling oocytes treated either by conventional IVF or ICSI is called mid-IVF/ICSI. We sum up here 487 attempts of this kind from six French ART centers. The mid-IVF/ICSI technique was performed in 5.6% of cases. The fertilization rate by micro-injected oocytes was significantly higher (P<0.01) than oocytes inseminated conventionally, 72.6% versus 53.4%. A failure of fertilization was observed only in mid-IVF in 21.6% of cases, which prevented a complete fertilization failure when we decided to propose to the couples concerned the mid-IVF/ICSI technique. Conversely, in 75.2% of cases, fertilization was found for the two batches of oocytes. The overall pregnancy rate has improved since the use of the mid-IVF/ICSI technique (33.1% versus 28.9%, P=0.013) and the fertilization failures decreased (10.4% versus 14.3%, P=0. 019). The pregnancy rate in only mid-IVF/ICSI cases is very high at 39.8% but for a selected population. The indications for mid-IVF/ICSI remain to be clarified especially with regard to male and idiopathic indications., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)

Vitrification with the use of "Open" carrier devices (Cryoloop, cryotop, cryoleaf, Vitriplug) which allowed the contact with liquid nitrogen has become a more popular way to achieve cooling rate superior to 20,000 °C/min. Even though the question of contamination with liquid nitrogen during ultra-rapid cooling and storage remain debatable with the use of "open" devices, it is important to revise the carrier system in a way, which minimizes the risk of contamination. According to the EU tissues and cells directive, it is advisable that the cooling and storage should be carried out in embryo carrier devices ensuring complete separation of the embryos from liquid nitrogen in a way, which minimizes the risk of contamination. The consequence of a reduction in the cooling rate resulting from the heat-insulating barrier of aseptic devices has to be counteracted by gradually increasing intracellular concentrations of cryoprotectants without inducing a toxic effect. We developed an aseptic vitrification method of vitrification for MII oocytes and embryos at different stage of development using the "VitriSafe" as "closed" carrier device., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)

All indicates that anti-Müllerian hormone (AMH), though initially studied for its importance on male development, plays an inhibiting role on the initial and cyclic processes of follicular recruitment. The aspects involved in its regulation are still poorly understood, but the oocyte, some steroids, and follicular development itself seem to be involved. In addition, AMH has become an important clinical marker of ovarian functioning for many reasons, including its exclusive production by granulosa follicles at many stages of development, its probable FSH independence, its low inter and intracycle variability and its reliable quantitative (qualitative?) relationship with ovarian follicles and their response to exogenous FSH. The growing interest in ovarian AMH incited us to review some important fundamental and clinical publications in this field., (Copyright 2010. Published by Elsevier SAS.)

Polycystic ovary syndrome (PCOS) is the most common etiology of menstrual disorders and hyperandrogenism. It is characterized by an excess of ovarian follicles. The mechanisms that underlie folliculogenesis disorder in PCOS appear to arise from primitive ovarian hyperandrogenism. This can be modulated by hormonal factors, such as LH or insulin. Ovarian hyperandrogenism results from a real theca cells dysfunction, whose origin is still poorly understood. It seems that complex genetic factors may be involved, but these have not yet been clearly identified. PCOS also results from granulosa cells dysfunction. For example, intra-ovarian factors, such as anti-mullerian hormone, are possibly involved in ovulation's disorders by blocking the physiological process of follicular recruitment. In turn, the oocyte could also be one of the actors possibly involved in the follicular excess in PCOS., (2010 Elsevier Masson SAS. All rights reserved.)

Folliculogenesis covers the sequential steps in the development of a follicle, from primordial to preovulatory. Most of the time, one follicle contains a single oocyte, but some follicles are polyovular in that they contain several. These follicles were considered earlier as pathological, but they are, actually, fairly common in several species, from mice to humans. The frequency of polyovular follicles (number of polyovular/total number of follicles) varies among species, <0.1% to 14% in the dog, and with age (more polyovular follicles during the prepubertal period). More than 20 oocytes (and even more than 100 in the marsupials like the opossum) may be present in a single follicle. These follicles may form during the earliest stages of follicle formation, due to an imbalance between somatic and germinal cells, which induces an incomplete germ cell cyst breakdown. In polyovular follicles, the quality (size and maturity) of the various oocytes is often heterogeneous. Numerous authors reported that polyovular follicles are able to reach ovulation and ovulate. Polyovular follicles, naturally found in several species, may also be induced by exposure to therapy or agents in the environment, especially with estrogen activity such as pesticides or diethylstilbestrol/DES. Polyovular follicles are also observed in the ovaries of mutated rodents., (2010 Elsevier Masson SAS. All rights reserved.)

Follicular fluid is a complex extracellular fluid, semi-viscous and yellow in colour, which accumulates in the antrum of ovarian follicles during their growing phase. Follicular fluid provides the microenvironment within which the cumulus-oocyte complex matures and granulosa cells differentiate. Scientists agree that follicular fluid derives mainly from plasma via the vascular compartment in the follicle wall. However, it also contains factors produced locally by the follicle cells, the production of which varies during different reproductive states. The aim of this paper is to review current knowledge on the formation, composition and roles of follicular fluid., (2010 Elsevier Masson SAS. All rights reserved.)

Only 5% of oocytes lead to life birth after IVF. Currently, it is difficult to evaluate oocyte quality on the sole basis of morphology criteria. Embryo morphology is considered as a good marker of potential implantation of embryo after ET, however it is not enough. Therefore, we need to develop non invasive assessment of oocyte quality. Research performed on follicular fluid may represent a new tool of increasing interest. The presence of G-CSF in follicular fluid could predict ongoing pregnancy. Cytokines, IL-7 and IL-17 could be also a predictor of success., (Copyright 2010 Elsevier Masson SAS. All rights reserved.)

From all the possibilities of assessment of the embryonic viability in vitro, morphology observed at microscope is the most elective criteria because of its specificity, sensibility and its armless side. The use of classifications at each stage of the development is essential for the selection of the right embryo to be replaced but do not allow dynamic evaluation. The development of computerized tools to capture and analyse images must emancipate us from oversimplification that doesn't cover frequent daily situations. The purpose is to examine the most frequent widespread classifications, the proposed alternatives and the consequences on embryonic replacement policy. More than a simple morphological factual evaluation of an attempt, it would be interesting to correlate embryonic development to stimulation criteria or quality culture media and other variables to enhance the obtaining of good prognostic embryos. The retrospective analysis of the databases of images will probably respond to this preoccupation.

Preimplantation embryo development is one of the key features with implantation itself to achieve a pregnancy. Assisted Reproductive Technologies both in human and animal have improved our knowledge on these events, although it remains elusive to predict embryo potential to give a baby. Among various ways to define embryo viability, noninvasive approaches get a serious advantage linked to the final transfer of the embryo. Techniques devoted to characterize the embryo secretome using proteomic or metabolomic approaches may be non invasive. Based on a direct identification of products of the embryo metabolism or an assessment of profile(s) related with embryo viability, they have greatly improved their sensitivity to allow their use in clinical embryology, once validated. Oocyte-cumulus dialogue, as a key factor for oocyte competence to meiosis and embryo development, was particularly concerned with both genomic and proteomic assessment of cumulus cells. While it is not possible to designate at the time being which among these approaches will be robust and cost-efficient enough to help routinely the clinical embryologist in assisted reproductive techniques (ART), one can predict that our ability to select the "right" embryo will combine morphological criteria already available with validated biomarkers.

Performance and security questions in human oocyte cryopreservation have been taking researchers for about two decades. Oocytes are usually frozen at metaphase II. Immature oocytes cryopreservation is still a research alternative. Two techniques are currently available for oocyte cryopreservation: slow freezing and vitrification. Experimental data suggest that vitrification has less impact on oocyte physiology than classical slow freezing. After slow freezing of mature oocytes, survival and fertilization rates reach 70 to 80% whereas cleavage rates are around 90%, leading to five implantations and 1.2 births per 100 thawed oocytes. After vitrification of mature oocytes, survival and cleavage rates reach 90% leading to 11 implantations and 1.8 births per 100 thawed oocytes. The obstetrical and neonatal prognosis of these pregnancies is reassuring. No increased risk of congenital anomalies has been observed. However, further evaluation is needed to guarantee the safety of cryopreservation procedures. Immature oocyte cryopreservation is not currently perfected but some indications appear of great interest.

Objective: To compare the oocyte and embryo quality and outcomes in polycystic ovary syndrome (PCOS) patients and normo-ovulatory women undergoing intracytoplasmic sperm injection (ICSI) cycles., Materials and Methods: We studied retrospectively 100 controlled ovarian stimulation cycles for ICSI in women with PCOS and 200 cycles in normo-ovulatory women. Every PCOS woman was paired according to age with two normo-ovulatory woman undergoing ICSI during the same study period. A long protocol of gonadotrophin releasing hormone (GNRH) agonist was carried out in all patients. The main outcome measures evaluated were: cancellation of the cycles, number of aspirated follicles, oocyte maturity, fertilization rate, embryo quality pregnancy and implantation rates, clinical abortion rate, multiple pregnancy rate and the "take-home baby" rate. Khi(2) test and Student t test were used for differences between normo-ovulatory and PCOS patients and the limit of significance was set at p<0,05., Results: ICSI was performed for male infertility in most cases (79% in PCOS group vs 79,5%; NS). There was no significant difference in term of cancellation rate (5,5% in PCOS group vs 5%; NS). The mean number of follicles was higher in patients with PCOS (18,1+/-8,5 vs 9,4+/-5,5; p<0,001). Oocyte mature rate (67% vs 52%; p<0,001), fertilization rate (75% vs 63,7%; p<0,001) and grade 1 embryo rate (69% vs 53%; p<0,001) were significantly higher in PCOS group. The mean number of transferred embryos was similar in the two groups (2,46+/-0,5 vs 2,54+/-1,01; NS). Implantation rate (16,6% vs 12,1%; NS), clinical pregnancy rate per transfer (31,5% vs 24%; NS) and Live birth rate (22% vs 20%; NS) did not differ statistically in the two groups. Twin and triplet pregnancies rates were similar in the two groups (20% vs 16,7%; NS and 6,6% vs 7,1% ; NS respectively). Miscarriage rate was higher in PCOS group but this did not reach the statistical significance (26,6% vs 16,6%; NS)., Conclusion: PCOS patients showed better global oocyte and embryo quality. However, pregnancy and live birth rates were similar in the two groups when number and quality of transferred embryos are equivalent.

The impact of nutrition and energy reserves on the reproductive functions is known for a very long time. However, the metabolic factors involved in the interactions between nutrition and reproduction are still poorly understood. These factors may be hormones or nutrients (glucose, protein and fatty acids). However, it remains to determine whether these factors act directly or indirectly on the reproductive tissues. In this issue, we briefly summarize the impact of fatty acids on the development of ovarian follicles, oocyte and embryo. We then discuss the current hypotheses about the mechanisms of action of these fatty acids on the ovarian functions. We describe more particularly the role of some receptors of fatty acids, Peroxisome Proliferator-Activated Receptors (PPAR) and Liver X Receptors (LXR) and two adipokines, leptin and adiponectin on ovarian cells.

Until the introduction of Assisted Reproductive Technologies (ART), many studies were conducted in order to evaluate their impact upon the children's health born in such a way. The epigenetic-risk notion was invoked and a link between ART and diseases associated with imprinting alterations was suggested with different examples, such as Beckwith-Wiedemann syndrome (BWS), Angelman syndrome (AS) and Silver-Russell syndrome (SRS). The epigenetic "life cycle" of imprinting (germline erasure, germline establishment, and somatic maintenance) concerns all the phases from gametogenesis, gamete maturation, fertilization, to early embryo development and appears particularly vulnerable to perturbations induced by superovulation, in vitro fertilization, embryo culture and embryo transfer. The studies, performed in model animal, provide a basis of the understanding of imprinting alterations induced by the ART and clinically useful information in order to improve the ART.

Preimplantation embryo development is one of the key features with implantation itself to achieve a pregnancy. Assisted Reproductive Technologies (ART) both in human and animal have improved our knowledge on these events, although predicting embryo potential to give a baby remains elusive. However data from the last 10 years have allowed either to hierarchize the available parameters or to open some new perspectives to predict embryo developmental potential. Both kinetics and morphological parameters belong to noninvasive quality embryo assessment for many years, although recent data on microvideographic analysis and multivariate analysis led to reduce their biological meaning. Moreover, new technical detection of meiotic spindle birefringency or zona pellucida anisotropy has improved the oocyte quality assessment with a deep impact for countries with restrictive legislation. Beyond such morphological criteria, more functional approaches concerned the oocyte (embryo) or its environment. Direct transcriptomic analysis, while invasive and therefore experimental, brought important data on embryo "quality". However, noninvasive metabolomic or proteomic analysis of embryo media gave promising results as well as respirometry. The environment of the oocyte has focused a specific attention, either based on regulatory proteins or cytokines present in follicular fluid, or involving genes or proteins from cumulus cells, as oocyte-cumulus dialog is a key factor in oocyte maturation. Whereas it is not possible for the time being to predict which parameter(s) will be implemented routinely, all data obtained underline that the ability to develop and implant is not based on embryo superlatives (more rapid, expressing more genes or proteins, larger metabolites uptake) but rather on a quiet state, as claimed by Leese some years ago, where a lot of resources would not be mobilized by any stressful situation.

Embryo quality is routinely evaluated based on morphological and kinematic criteria, but it should also be founded on clinical and biological data from the ovulation induction treatments and the data from the SPZ test, with, if necessary, their genetic analysis. Zygote quality evaluation is based on the zygote score or the P0 profiles. Embryo cleavage kinetics is evaluated at D1 (early cleavage 25-27 h post-insemination), at D2 (four cells), and at D3 (eight cells), and their selection, particularly for eSET, is for the most part based on the cell number and regularity, the percentage of fragmentation (< or = 20 %) and the location of the fragments and the absence of blastomere polynucleation. Imaging software (Fertimorph) now facilitates the evaluation of embryo quality.

The main advantage of freezing oocytes is preserving the future fertility of women whose ovarian function is threatened by medical or surgical treatments and facilitating oocyte donation. The results of slow freezing of oocytes seem relatively dependent on the protocols used, most particularly on the sucrose concentration in the solutions of cryoprotective agents used. Today it seems that the use of low or medium-level concentrations of sucrose during the freezing process gives better results in terms of pregnancies and implantation than higher concentrations (0.3 mol/l), despite the sometimes less favorable results on oocyte survival after thawing. The cumulated pregnancy rates from frozen/thawed oocyte transfer are nearly equivalent to the results observed with frozen/thawed embryo transfer, which makes oocyte cryopreservation a valid alternative to embryo cryopreservation. Moreover, it seems that these results can be improved by the vitrification technique.

Objectives: Since we observed in our laboratory a significant decrease of the fertilization rate in the classical IVF procedure compared to ICSI, we wanted to evaluate a new insemination procedure, using sperm morphology of selected semen as a criterion taken into account to calculate the number of mobile sperm to inseminate., Patients and Methods: Therefore, we designed a prospective autocontrolled study which took place from September 2006 to May 2007. All IVF attempts resulting from infertile couples were included in the study if a previous analysis (lesser than one year) of sperm parameters performed in our laboratory was assessed as normal. The retrieved oocytes were separated into two groups (sibling ovocytes), inseminated randomly either with the usual procedure (control group) or with the tested protocol (tested group). Fertilization rate was the primary end point to evaluate this assay. Polyspermy rates were also compared. Subsequently, embryonic quality and development were assessed to eliminate an oxidative stress impact. Paired Student t-test was applied for statistical analysis., Results: In all, 130 couples were included. The diploid fertilization rate was significantly increased in the tested group compared to the control group (66.9% vs 61.3%; p=0.017). No statistically significant difference was showed between the polyspermy rate and numbers of (i) good quality embryos and (ii) blastomeres per embryo into the two groups., Discussion and Conclusion: Compared with the conventional insemination procedure, the fertilization rate benefited from the tested insemination method in our laboratory without damaging embryonic growth. In the limit of our procedure, selected sperm morphology should be used to inseminate an appropriate number of spermatozoa within oocytes, even in case of normal semen values.

In Assisted Reproductive Technology (ART), the pregnancy and birth rates following in vitro fertilization (IVF) attempts are still low. Recently, research in the field of ART has explored new oocyte and embryo quality selection criteria such as apoptotic markers. Many studies provided evidence that bad oocyte and embryo quality can be associated with apoptosis. The aim of this review is to summarize our current knowledge on the apoptosis process in oocytes and embryos, and focus on the possibility of using apoptotic markers as a reliable and predictive marker to select competent oocytes and embryos during IFV.