Is in vitro maturation of oocytes retrieved ex vivo from ovarian tissue an effective fertility preservation technique in the presence of organic ovarian cysts?

OBJECTIVE(S): In vitro maturation (IVM) of oocytes retrieved ex vivo from ovarian tissue (OTO-IVM) could be an additional source of mature oocytes with the potential to optimise medical fertility preservation (FP) after oophorectomy. It is often undertaken at the same time as the ovarian tissue cryopreservation (OTC). In the presence of an organic ovarian cyst, OTO-IVM could prove to be the only technique available to permit FP since ovarian stimulation, transvaginal ovarian needle puncture or future ovarian tissue graft are contraindicated. However, the presence of an organic cyst could alter follicular growth and the number of retrievd oocytes. Our study aims to assess the efficiency of OTO-IVM in such situations. STUDY DESIGN: Retrospective, observational study involving 20 female patients with FP by OTO-IVM between May 2017 and November 2021 at the University Hospital of Toulouse. Oocytes retrieved "ex vivo" were transferred to an IVM medium with HP-hMG, LH and HSA and then vitrified after 24 to 48 h of IVM. Data analysis was performed on the total population and comparatively between patients who had or did not have an organic ovarian cyst. RESULTS: The indications included 15 oncologic and 5 non-oncologic indications. Ten had an organic ovarian cyst on the retrieved ovary. The number of retrieved oocytes was 17.4+/-12.0 in the absence of cyst vs 4.1+/-6.3 in the presence (p = 0.003). The number of vitrified mature oocytes was 5.8+/-5.3 in the absence vs 1.1+/-2.2 (median = 0) in the presence of a cyst (p = 0.03). Ninety percent of the patients with an organic cyst had less than two vitrified mature oocytes. The mean maturation rate was 34%, not significantly different between the two groups. We found a correlation between serum AMH level and the number of mature oocytes: ρ:0.47 CI95 = [0.02; 0.76]; p = 0.04. CONCLUSION(S): OTO-IVM is an additional source of mature oocytes to optimise FP after oophorectomy. However, in the presence of an organic ovarian cyst on the retrieved ovary, the exocrine, paracrine and endocrine functions of the ovary are impaired. As such, the number of immature oocytes obtained is highly impacted and appears to be insufficient to be able to propose systematically this technique in such situations.

Pre- and Postnatal Dietary Exposure to a Pesticide Cocktail Disrupts Ovarian Functions in 8-Week-Old Female Mice.

Female infertility has a multifactorial origin, and exposure to contaminants, including pesticides, with endocrine-disrupting properties is considered to be involved in this reproductive disorder, especially when it occurs during early life. Pesticides are present in various facets of the environment, and consumers are exposed to a combination of multiple pesticide residues through food intake. The consequences of such exposure with respect to female fertility are not well known. Therefore, we aimed to assess the impact of pre- and postnatal dietary exposure to a pesticide mixture on folliculogenesis, a crucial process in female reproduction. Mice were exposed to the acceptable daily intake levels of six pesticides in a mixture (boscalid, captan, chlorpyrifos, thiacloprid, thiophanate and ziram) from foetal development until 8 weeks old. Female offspring presented with decreased body weight at weaning, which was maintained at 8 weeks old. This was accompanied by an abnormal ovarian ultrastructure, a drastic decrease in the number of corpora lutea and progesterone levels and an increase in ovary cell proliferation. In conclusion, this study shows that this pesticide mixture that can be commonly found in fruits in Europe, causing endocrine disruption in female mice with pre- and postnatal exposure by disturbing folliculogenesis, mainly in the luteinisation process.

Semi-automated versus manual embryo vitrification: inter-operator variability, time-saving, and clinical outcomes.

PURPOSE: Does semi-automated vitrification have lower inter-operator variability and better clinical outcomes than manual vitrification? METHODS: Retrospective analyses of 282 patients whose embryos had been cryopreserved, manually with Irvine®-CBS® (MV) or semi-automatically vitrified with the GAVI® method (AV) (from November 2017 to September 2020). Both techniques were performed during the same period by 5 operators. Inter-operator variability was statistically analyzed between operators who performed the vitrification and those who performed the warming process to compare the intact survival rate (% embryos with 100% intact blastomeres) and the positive survival rate (at least 50% intact blastomeres). Additionally, the complete vitrification time was assessed for the 2 techniques according to the number of vitrified embryos. RESULTS: Manual vitrification involved warming 338 embryos in 266 cycles for 181 couples compared to 212 embryos in 162 AV cycles for 101 patients. The positive survival rate was higher (p < 0.05) after MV (96%; 323/338) than after AV (90%; 191/212). The intact survival rate (86 vs 84%) and the clinical pregnancy rate (27 vs 22%) were not significantly different between MV and AV. Regarding the inter-operator variability, no significant difference in positive and intact survival rate was evident between the 5 technicians, neither by vitrification nor by warming steps with MV and AV. Concerning time-saving, the MV technique proved to be quicker than AV (minus 11 ± 9 min). CONCLUSIONS: Manual vitrification exhibited favorable total survival rates and was more time efficient, while both MV and AV cooling and warming treatments showed little operator variability.

Need for choosing the ideal pH value for IVF culture media.

PURPOSE: Monitoring the pH of IVF culture media is a good practice, but the required pH levels have been "arbitrarily" set. Assisted reproductive technology centers around the world are spending time and money on pH monitoring without any consensus to date. The objective of this narrative review was to evaluate the importance of pH monitoring during IVF, discover how the oocyte and embryo regulate their intracellular pH and try to determine the optimal pH to be applied. METHODS: A narrative literature review was performed on publications in the PubMed database reporting on the impact of pH on cellular function, oocyte and embryo development, IVF outcomes and pathophysiology, or on physiological pH in the female reproductive tract. RESULTS: Intracellular pH regulates many cellular processes such as meiotic spindle stability of the oocyte, cell division and differentiation, embryo enzymatic activities, and blastocoel formation. The internal pH of the human embryo is maintained by regulatory mechanisms (mainly Na+/H+ and HCO3-/Cl- exchangers) that can be exceeded, particularly in the oocyte and early-stage embryos. The opinion that the optimal pH for embryo culture is physiological pH is not correct since several physicochemical parameters specific to IVF culture conditions (temperature, medium composition, duration of culture, or implication of CO2) can modify the intracellular pH of the embryo and change its needs and adaptability. CONCLUSIONS: Because correct and stable extracellular pH is essential to embryo health and development, monitoring pH is imperative. However, there is a lack of clinical data on choosing the ideal pH for human IVF culture media.

Anti-sperm antibodies detection by a modified MAR test: Towards a better definition of its indications.

RESEARCH QUESTION: Anti-sperm antibodies (ASA) have been shown to reduce male fertility but consensus about the precise situations in which tests should be carried out are lacking. In infertility investigations, should the mixed antiglobulin reaction (MAR) test be a first-line test? Should it be carried out systematically before assisted reproductive technology (ART)? What are the risk factors for ASA? DESIGN: All infertile patients (n = 1364) were tested with SpermMar (modified MAR test) between July 2013 and June 2017. Intra-patient variability of the MAR test was also assesed by comparing two tests within the same year in selected patients (n = 101). RESULTS: The main factor that influenced the percentage of ASA was the presence or absence of sperm agglutination. In the presence of agglutinations, 27 out of 72 (37.5%) patients were positive for ASA compared with 33 out of 1292 (2.6%) in the absence of agglutinations (P < 0.0001). When one risk factor was present (spontaneous sperm agglutination, history of scrotal trauma or inguinal surgery), 33 out of 179 (18.44%) tests were positive for ASA (≥50% coated spermatozoa), whereas only 27 out of 1242 (2.2%) were positive when no risk factor was present (P < 0.0001). CONCLUSIONS: ASA detection should not be systematically recommended in investigations of fertility status and before ART but reserved for when sperm agglutination is found during conventional sperm examination, or if the patient has a history of scrotal trauma or has undergone inguinal surgery.

Environmental pollutants, a possible etiology for premature ovarian insufficiency: a narrative review of animal and human data.

BACKGROUND: Because only 25% of cases of premature ovarian insufficiency (POI) have a known etiology, the aim of this review was to summarize the associations and mechanisms of the impact of the environment on this pathology. Eligible studies were selected from an electronic literature search from the PUBMED database from January 2000 to February 2016 and associated references in published studies. Search terms included ovary, follicle, oocyte, endocrine disruptor, environmental exposure, occupational exposure, environmental contaminant, pesticide, polyaromatic hydrocarbon, polychlorinated biphenyl PCB, phenol, bisphenol, flame retardant, phthalate, dioxin, phytoestrogen, tobacco, smoke, cigarette, cosmetic, xenobiotic. The literature search was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We have included the human and animal studies corresponding to the terms and published in English. We have excluded articles that included results that did not concern ovarian pathology and those focused on ovarian cancer, polycystic ovary syndrome, endometriosis or precocious puberty. We have also excluded genetic, auto-immune or iatrogenic causes from our analysis. Finally, we have excluded animal data that does not concern mammals and studies based on results from in vitro culture. Data have been grouped according to the studied pollutants in order to synthetize their impact on follicular development and follicular atresia and the molecular pathways involved. Ninety-seven studies appeared to be eligible and were included in the present study, even though few directly address POI. Phthalates, bisphenol A, pesticides and tobacco were the most reported substances having a negative impact on ovarian function with an increased follicular depletion leading to an earlier age of menopause onset. These effects were found when exposure occured at different times throughout the lifetime from the prenatal to the adult period, possibly due to different mechanisms. The main mechanism seemed to be an increase in atresia of pre-antral follicles. CONCLUSION: Environmental pollutants are probably a cause of POI. Health officials and the general public must be aware of this environmental effect in order to implement individual and global preventive actions.

How many embryos should be transferred? A validated score to predict ongoing implantation rate.

OBJECTIVE: To developed a prognostic score to predict ongoing implantation rate according to clinical and biological parameters and to choose the number of embryos to be transferred in patients undergoing IVF/ICSI. STUDY DESIGN: The transfer score was established using multivariate analysis of biological and clinical parameters in 3211 fresh embryo transfers in a retrospective study. Then we validated the score in 694 fresh embryo transfers in a prospective study. We assessed ongoing implantation rates, ongoing pregnancy rates, multiple pregnancy rates and live birth rate. RESULTS: Among the different variables tested, 4 were identified that influenced the implantation rate: female age, the ratio of retrieved oocytes/mean daily dose of injected FSH, attempt rank and the morphology of the embryo cohort. Prospective application of this score resulted in significantly lower number of transferred embryos (1.8 vs 2.0 P<0.001) and lower twins rates (9.7% vs 17.3%, P<0.001) without decreasing live birth rates. CONCLUSION: Although the risks of multiple pregnancies should not be ignored, it appears excessive to impose the limit of 2 embryos for transfer particularly in situations with a poor prognosis. We sought to provide a personalized prognosis by using clinical and embryo data in order to choose the number of embryo(s) for transfer with a moderate multiple pregnancy rate of less than 11%.

Continued existence of significant disparities in the technical practices of sperm morphology assessment and the clinical implications: results of a French questionnaire.

OBJECTIVE: To investigate current practices for sperm morphology assessment. DESIGN: E-mail survey questionnaire. SETTING: Diagnostic and clinical institutions/laboratories. PATIENT(S): French biologists and clinicians (n = 225). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Answers to 24 questions. RESULT(S): The survey shows that even now in France there is great inconsistency in the use of sperm morphology assessments. For example, the survey revealed that no fewer than six different staining techniques were in use. Automatic reading is hardly used (used by 2.5% of the biologists replying to the survey). More than 33.6% of biologists use a threshold of normal forms unsuited to their classification, and 20% do not perform any internal quality control in this area. Prescribing doctors seldom trust the tests, likely due to their lack of analytic reliability. Among the biologists surveyed 26% said the percentage of normal forms is either unreliable or not very reliable in analytic terms, and 24% of clinicians stated that it has little clinical relevance. CONCLUSION(S): The survey reveals a marked lack of uniformity in French laboratories for performing sperm morphology assessment and in the use of the results by physicians. Regular quality control procedures and well-trained personnel, up to date with their training and conversant with the latest techniques as well as harmonized practices, are clearly indispensable. It is time for a consensus on the practice and interpretation of this particular test.