In Vitro Fertilization Techniques Research Articles

Advanced Sperm Selection Techniques for Assisted Reproduction

: Male infertility accounts for approximately 40% of infertility cases. There are many causes of male infertility, including environmental factors, age, lifestyle, infections, varicocele, and cancerous pathologies. Severe oligozoospermia, cryptozoospermia, and azoospermia (obstructive and non-obstructive) are identified as severe male factor infertility, once considered conditions of sterility. Today, in vitro fertilization (IVF) techniques are the only treatment strategy in cases of male factor infertility for which new methodologies have been developed in the manipulation of spermatozoa to achieve fertilization and increase success rates. This review is an update of in vitro manipulation techniques, in particular sperm selection, emphasizing clinical case-specific methodology. The success of an IVF process is related to infertility diagnosis, appropriate choice of treatment, and effective sperm preparation and selection. In fact, selecting the best spermatozoa to guarantee an optimal paternal heritage means increasing the blastulation, implantation, ongoing pregnancy and live birth rates, resulting in the greater success of IVF techniques.

Advanced Sperm Selection Techniques for Assisted Reproduction.

Male infertility accounts for approximately 40% of infertility cases. There are many causes of male infertility, including environmental factors, age, lifestyle, infections, varicocele, and cancerous pathologies. Severe oligozoospermia, cryptozoospermia, and azoospermia (obstructive and non-obstructive) are identified as severe male factor infertility, once considered conditions of sterility. Today, in vitro fertilization (IVF) techniques are the only treatment strategy in cases of male factor infertility for which new methodologies have been developed in the manipulation of spermatozoa to achieve fertilization and increase success rates. This review is an update of in vitro manipulation techniques, in particular sperm selection, emphasizing clinical case-specific methodology. The success of an IVF process is related to infertility diagnosis, appropriate choice of treatment, and effective sperm preparation and selection. In fact, selecting the best spermatozoa to guarantee an optimal paternal heritage means increasing the blastulation, implantation, ongoing pregnancy and live birth rates, resulting in the greater success of IVF techniques.

O-091 Severe maternal and neonatal morbidity among gestational carriers: a population-based cohort study

Abstract Study question Are gestational carriers at higher risk of severe maternal (SMM) or neonatal (SNM) morbidity, compared to patients using unassisted conception or in vitro fertilization (IVF)? Summary answer A higher risk of SMM and adverse pregnancy outcomes is seen among gestational carriers, without evident neonatal morbidity. What is known already The use of gestational carriers (GC) also known as “surrogate pregnancy” is increasing. Gestational carriage may be associated with adverse outcomes linked to both IVF techniques and exposure to foreign fetal antigens. Adverse outcomes of GC pregnancies are largely unknown. Study design, size, duration Retrospective population-based cohort study of 937,938 births, derived from existing administrative health datasets. Data were linked using unique for birthing woman and child identifiers and analyzed at ICES Ontario. The study excluded participants with untreated infertility, those utilizing ovulation induction or intrauterine insemination, and those with multifetal gestation. Participants/materials, setting, methods Singleton births at > 20 weeks gestation in Ontario, 2012-2021. Pregnancy characteristics were obtained from the Ontario Birth Registry database. Exposure groups were i) unassisted conception (comparison group 1), ii) IVF (comparison group 2), or iii) GC. Primary outcomes included validated composite outcomes of i) SMM and ii) SNM. Secondary outcomes included hypertensive disorders of pregnancy, cesarean delivery, preterm birth, and postpartum hemorrhage. Modified Poisson regression models generated relative risks adjusted for birthing woman’s characteristics (aRR). Main results and the role of chance Of all singleton births, 937,938 (97.7%) were from unassisted conception, 20,958 (2.2%) from IVF, and 956 (0.1%) from GC. GC were more likely to be parous, reside in a lower-income area, and have higher rates of chronic hypertension. GC were more likely than unassisted conception women to be older and be non-smokers, with some opposite trends when compared to IVF recipients. Respective rates of SMM were 2.4%, 4.6% and 7.1%. The aRR were 3.07 (95% CI 2.44-3.87) comparing GC to unassisted conception, and 1.88 (95% CI 1.46-2.42) comparing GC to IVF. Respective rates of SNM were 6.0%, 9.1% and 6.5%, generating aRR of 1.21 (95% CI 0.95-1.54) for GC vs. unassisted conception, and 0.78 (95% CI 0.60-1.00) for GC vs. IVF. Respective rates of hypertensive disorders were 6.6%, 11.6% and 13.9%. The aRR were 1.89 (1.61 - 2.21) comparing GC to unassisted conception, 1.42 (1.19 - 1.69) comparing GC to IVF. Respective rates of postpartum hemorrhage were 5.7%, 10.5% and 13.9%, generating aRR of 2.87 (2.43 - 3.38) for GC vs. unassisted conception, and 1.40 (1.17 - 1.67) for GC vs. IVF. Limitations, reasons for caution The databases did not include certain baseline details, including the reasons for opting for a GC and the sources of gametes. There is a chance that the measured association may be distorted or confounded by the unmeasured variables. Wider implications of the findings GC appear to be at an elevated risk of maternal morbidity, without evident risk of neonatal morbidity. Further studies are needed to understand the mechanisms involved. Eligibility for GC should be determined using consistent selection criteria, and consideration should be given to specialized obstetrical care to promote optimal pregnancy outcomes. Trial registration number Not applicable

Multiple births from medically assisted reproduction: contribution of different types of procedures and trends over time

ObjectiveTo evaluate the relative impact of different strategies of medically assisted reproduction (MAR), i.e. first line treatment (ovarian stimulation with or without intrauterine insemination) and in vitro fertilization (IVF) procedures (conventional IVF or intracytoplasmic sperm injection), on the risk of multiple births. Study designWe utilized the health care utilization databases of the Lombardy region to identify births resulting from MAR between 2007 and 2022. We gathered data on the total number of multiple births and calculated the prevalence rate by dividing the number of multiples by the total number of births. To examine the temporal trend in the proportion of multiple births after MAR over time, a linear regression model was employed separately for different types of techniques and in strata of maternal age. ResultsA total of 30,900 births after MAR were included; 4485 (14.5 %) first line treatments and 26,415 (85.5 %) IVF techniques. Overall, 4823 (15.6 %) multiple births were identified. The frequency of multiple births over the study period decreased from 22.0 % in 2007 to 8.7 % in 2022 (p < 0.01). Multiple births from first line treatments were stable ranging from 13.5 % in 2007–2008 to 12.0 % in 2021–2022 (p = 0.29). Multiple births from IVF procedures decreased from 23.8 % in 2007–2008 to 8.4 % in 2021–2022 (p < 0.01). Stratifying for maternal age (i.e. < 35 and ≥ 35 years), the trends remained consistent. ConclusionsThe reduction in multiple births has been influenced by changes in IVF strategy and procedures. The decline has been gradual but steady since 2009, when a law restricting embryo freezing was repealed in Italy. In contrast, the proportion of multiple births resulting from first line treatments has remained constant over time. Despite declining, multiple births from MAR remained about one order of magnitude higher than those from spontaneous pregnancies.

Deep Learning-Based Detection of Human Blastocyst Compartments with Fractal Dimension Estimation

In vitro fertilization (IVF) is an efficacious form of aided reproduction to deal with infertility. Human embryos are taken from the body, and these are kept in a supervised laboratory atmosphere during the IVF technique until they exhibit blastocyst properties. A human expert manually analyzes the morphometric properties of the blastocyst and its compartments to predict viability through manual microscopic evaluation. A few deep learning-based approaches deal with this task via semantic segmentation, but they are inaccurate and use expensive architecture. To automatically detect the human blastocyst compartments, we propose a parallel stream fusion network (PSF-Net) that performs the semantic segmentation of embryo microscopic images with inexpensive shallow architecture. The PSF-Net has a shallow architecture that combines the benefits of feature aggregation through depth-wise concatenation and element-wise summation, which helps the network to provide accurate detection using 0.7 million trainable parameters only. In addition, we compute fractal dimension estimation for all compartments of the blastocyst, providing medical experts with significant information regarding the distributional characteristics of blastocyst compartments. An open dataset of microscopic images of the human embryo is used to evaluate the proposed approach. The proposed method also demonstrates promising segmentation performance for all compartments of the blastocyst compared with state-of-the-art methods, achieving a mean Jaccard index (MJI) of 87.69%. The effectiveness of PSF-Net architecture is also confirmed with the ablation studies.

Micro/Nanorobotics in In Vitro Fertilization: A Paradigm Shift in Assisted Reproductive Technologies.

In vitro fertilization (IVF) has transformed the sector of assisted reproductive technology (ART) by presenting hope to couples facing infertility challenges. However, conventional IVF strategies include their own set of problems such as success rates, invasive procedures, and ethical issues. The integration of micro/nanorobotics into IVF provides a prospect to address these challenging issues. This article provides an outline of the use of micro/nanorobotics in IVF specializing in advancing sperm manipulation, egg retrieval, embryo culture, and capacity future improvements in this swiftly evolving discipline. The article additionally explores the challenges and obstacles associated with the integration of micro/nanorobotics into IVF, in addition to the ethical concerns and regulatory elements related to the usage of advanced technologies in ART. A comprehensive discussion of the risk and safety considerations related to using micro/nanorobotics in IVF techniques is likewise presented. Through this exploration, we delve into the core principles, benefits, challenges, and potential impact of micro/nanorobotics in revolutionizing IVF procedures and enhancing affected person outcomes.

When the Embryo Meets the Endometrium: Identifying the Features Required for Successful Embryo Implantation.

Evaluation of the optimal number of embryos, their quality, and the precise timing for transfer are critical determinants in reproductive success, although still remaining one of the main challenges in assisted reproduction technologies (ART). Indeed, the success of in vitro fertilization (IVF) treatments relies on a multitude of events and factors involving both the endometrium and the embryo. Despite concerted efforts on both fronts, the overall success rates of IVF techniques continue to range between 25% and 30%. The role of the endometrium in implantation has been recently recognized, leading to the hypothesis that both the "soil" and the "seed" play a central role in a successful pregnancy. In this respect, identification of the molecular signature of endometrial receptivity together with the selection of the best embryo for transfer become crucial in ART. Currently, efforts have been made to develop accurate, predictive, and personalized tests to identify the window of implantation and the best quality embryo. However, the value of these tests is still debated, as conflicting results are reported in the literature. The purpose of this review is to summarize and critically report the available criteria to optimize the success of embryo transfer and to better understand current limitations and potential areas for improvement.

Efficient and repeatable in vitro fertilization in rabbits

Rabbits constitute an interesting model to understand gamete interaction and test novel Artificial Reproductive Techniques, but in vitro fertilization (IVF) is particularly problematic in this species. We have conducted a series of experiments to develop a consistent IVF technique. Initially, we checked viability, acrosome integrity, capacitation and motility in ejaculated sperm purified by a density gradient and incubated at different times in three different media: Tyrode's Albumin Lactate Pyruvate (TALP), human tubal fluid (HTF), and Brackett and Oliphant (BO). Total and progressive motility at 10–24 h and linearity from 3 h onwards was significantly higher in BO medium compared to TALP and HTF. Subsequently, cumulus-oocyte complexes (COCs) collected 10 h after induction of ovulation were incubated with sperm in TALP, HTF or BO for 18 h with or without performing sperm pre-incubation for 6 h. Pronuclear formation rate at 18 h was significantly higher in BO compared to other media (∼84 % vs. 17–22 %) and was not improved by pre-incubation. As COCs recovery rate was low at 10 h after induction of ovulation, COCs were collected at 12 h and co-incubated with sperm in BO. Pronuclear formation rate was similar than those obtained in COCs collected at 10 h (∼85 %), and when embryos were allowed to develop in vitro, the protocol yielded high cleavage and blastocyst rates (91 and 59 %, respectively). In conclusion, ejaculated rabbit sperm purified in a density gradient fertilize efficiently COCs collected at 12 h in BO medium.

Evaluation of medical equipment of the infertility treatment centers in North western of Libya

Background Cell culture, particularly in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) are both of which assisted reproductive technology (ART) approaches that are most frequently utilized to treat infertility around the world. Objective Highlight the differences and similarities in instruments and equipment in ART laboratories that govern their success, limitations, and potential differences between laboratories by identifying the facilities of laboratories to improve outcomes of ICSI and IVF techniques in clinical centers on the western coast of Libya. Method: descriptive study, comparison of five (5) assisted reproduction laboratories instrument and equipment parameters using the same quality-control application in clinical centers on the western coast of Libya.Result: Among the seven clinical centers specializing in the diagnosis and treatment of infertility that were contacted, four infertility treatment centers and the Medical Research Center (MRC) provided a response. The MRC and four clinical centres had good laboratory facilities for ICSI and IVF techniques compared to Jordan ART laboratory facilities.Conclusion: This study highlights the imperative of enhancing laboratory infrastructure to support optimal implementation of ICSI and IVF procedures.

Carbon Monoxide Exposure Does Not Improve The In Vitro Fertilization Rate of Oocytes Obtained from Heterozygous Hmox1 Knockout Mice.

In our experimental study we explored the impact of maternal reduced heme oxygenase-1 (HO-1) gene (Hmox1) expression on the in vitro fertilization (IVF) rate through the use of heterozygous Hmox1 knockout mice models (HET/Hmox1+/ -). Also, we hypothesized a beneficial role of gametes exposure during fertilization to carbon monoxide (CO), one of HO-1 by-products, that might be relevant for the improvement of IVF rates. IVF technique was performed by using oocytes obtained from wild-type (WT) or Hmox1+/ - dams fertilized with WT, Hmox1+/ - or Hmox1-/ - mice-derived sperm. The fertilization step occurred either in a conventional incubator (37°C, 5% CO2) or in an incubator implemented with CO (500 ppm). The superovulation yield of WT and Hmox1+/ - mice and the number of fertilized oocytes was assessed using an optical microscope. The dams' Hmox1 heterozygous knockout neither impact the superovulation yield, nor did influence the fertilization success rate. Moreover, CO exposure during fertilization could not significantly improve the outcome. Our study showed that the maternal Hmox1+/ -condition is not affecting the IVF rate in mice. Furthermore, we discovered that CO exposure cannot be exploited to ameliorate this critical step of the IVF protocol.

Production of transgenic first filial puppies expressing mutated human amyloid precursor protein gene.

Propagation of transgenic animals by germline transmission using assisted reproductive technologies such as in vitro fertilization (IVF) is the most efficient way to produce transgenic colonies for biomedical research. The objective of this study was to generate transgenic puppies from a founder dog expressing the mutated human amyloid precursor protein (mhAPP) gene. Experiment I assessed the characteristics of the semen prepared by freshly diluted, swim-up, and Percoll gradient methods using a computer-assisted semen analyzer (CASA). Motile and progressively motile sperm counts were higher in the Percoll gradient samples (p < 0.05) than in the swim-up and freshly diluted samples. In Experiment II, a total of 59, 70, and 65 presumptive zygotes produced by fresh, Percoll gradient, and swim-up methods, respectively, were transferred to surrogates (5 for each group); the Percoll gradient (27.27%) and swim-up samples (14.29%) showed the highest blastocyst formation rates, while fresh diluted semen did not produce any blastocyst. Experiment III examined the full-term developmental ability of embryos. Among the 5 surrogates in the Percoll gradient group, one (20.0%) became pregnant; it had 4 (6.15%) sacs and delivered 4 (6.15%; 2 males and 2 females) live puppies. Among the 4 puppies, 2 (50.0%) were found to transmit the transgene on their nail and toe under GFP fluorescence. Furthermore, the integration and expression of the mhAPP transgene were examined in the umbilical cords of all the IVF-derived puppies, and the presence of the transgene was only observed in the GFP-positive puppies. Thus, semen prepared by the Percoll method could generate transgenic puppies by male germline transmission using the IVF technique. Our result will help propagate transgenic dogs efficiently, which will foster human biomedical research.

Cell-free DNA in Embryo Culture Media as Non-invasive Biomarker of the Quality of Embryo Cleavage

Background: The success of in vitro-fertilization (IVF) cycles is determined in large part by the quality of embryo cleavage, which in turn, is dependent on the quality of the embryo culture media (CM). Many factors can influence the quality of embryo CM, one of which is the levels of Cell Free Deoxyribonucleic acid (DNA). Understanding the association between Cell-free DNA levels in embryo CM and the quality of embryo cleavage could help improve the quality of IVF techniques.&#x0D; Methods: This prospective study was conducted with 96 spent CM from patients undergoing IVF cycle, in order to determine relationships of Cell-free DNA levels in embryo CM with embryo cleavage quality on day 3. After intracytoplasmic sperm injection (ICSI), 48 embryos were evaluated on day 3 of their development, according to their cell number. Day 2 and day 3 CM corresponding to each one of the embryos was analyzed, by quantitative PCR, for estimation of Cell-free DNA levels.&#x0D; Results: The results revealed a significant increase in Cell-free DNA levels on day 2 CM corresponding to 4 to 6 cell embryos compared to those corresponding to 7 to 8 cell embryos (p=0.04). As for day 3 CM, the results showed no significant difference between the Cell-Free DNA levels in CM of 7-8 and those of 4-6 cell embryos (p=0.4). Also, cell free DNA levels in embryo CM, were significantly higher on day 2 compared to day 3 for both 7-8 and 4-6 cell embryos (p=0.03; p=0.04). &#x0D; Conclusion: We conclude that cell-free DNA levels in CM might be associated with delayed embryo cleavage.

Hysteroscopic versus laparoscopic management in patients with communicating hydrosalpinx and planning for IVF: a randomized controlled trial

BackgroundHydrosalpinx is considered one of the obstacles that could hinder the success of IVF techniques due to the toxic effect of the hydrosalpinx fluid pouring into the uterine cavity. Tubal disconnection by either hysteroscopic or laparoscopic approaches is considered the standard in the operative management of hydrosalpinx prior to IVF cycles. The aim of the current study was to compare the success rate of hysteroscopic tubal occlusion using electro-coagulation with laparoscopic tubal disconnection in cases of hydrosalpinx prior to in vitro fertilization (IVF).MethodsA total of 108 women with unilateral or bilateral hydrosalpinx, who were candidates for tubal occlusion before IVF, were equally randomized into two groups. Group A (n = 54) underwent hysteroscopic tubal occlusion using the roller-ball electro-coagulation of the interstitial part of the tube and the uterine cornual area, and group B (n = 54) underwent laparoscopic tubal disconnection using bipolar coagulation and a proximal tubal cut. Operative time, complications, postoperative pain measured by the visual analogue score (VAS score), and postoperative hospital stay were recorded for both groups. The success rate of tubal occlusion was assessed 1 month later using a post-menstrual hysterosalpingogram (HSG).ResultsLaparoscopic tubal disconnection was more successful than the hysteroscopic approach regarding tubal occlusion rate (96.15% vs. 86.67% respectively, p = 0.044). The operative time and postoperative pain VAS scores in the hysteroscopy group (3.65 ± 1.03 min and 1.81 ± 1.35, respectively) were significantly lower than that in the laparoscopy group (17.48 ± 4.70 min and 4.06 ± 1.65, respectively) with p < 0.001.ConclusionAlthough laparoscopic tubal disconnection is more successful, the hysteroscopic approach is an alternative which has its own limitations that can be assessed by hysterosalpingogram, especially when laparoscopy is contraindicated, technically difficult, or refused by the patient.Trial registrationIt was first registered at ClinicalTrials.gov on 30/07/2019 with registration number NCT04037813.

Knowledge, Attitude, and Perceptions about In Vitro Fertilization (IVF) among Women of Childbearing Age in Cape Coast, Ghana.

Background Infertility impacts a lot of considerable negative social effects on the lives of infertile partners, especially females, who repeatedly experience the emotional sequelae of childlessness. The study's goal was to assess women of reproductive ages' awareness of IVF treatments, as well as their attitudes and misunderstandings about them in Cape Coast, Ghana. Methods A total of 437 reproductive-age women in Cape Coast Metropolis were recruited using a simple random sampling approach for this Cross-Sectional Descriptive study. Data were collected with a semistructured interviewer-administered questionnaire and were analyzed using IBM SPSS version 26.0, and p ≤ 0.05 was considered significant. Results The mean age was calculated to be 25.33 ± 0.066 years with a greater proportion, 65.7% within 15–24 years, 76.5% had no or had never had a child before. 93.4% were of the Christian faith, 66.8% were aware of IVF, and 74.8% think IVF offers hope. Although 41.4% believe it is not a natural procedure and 44.6% believe IVF children are normal but not natural. While 72.1% believe the treatment is very costly, and 40.7% believe it is not affordable or accessible. The majority believe IVF kids are legitimate (76.9%), and so should be welcomed by society (86.5%). The overall opinion of IVF service acceptability was 81.7% good. Seventy-two and three percent did not know whether IVF services are available in Cape Coast. Also, 48.1% were aware that IVF may result in pregnancy failure, with fewer than half (43.5%) believing it could be linked to genetic problems in the baby. The majority (60.4%) were willing to use IVF services, and 82.8% will utilize just their husband's sperm technique. While others may not want to undergo any form of IVF technique because they desire to conceive naturally (51.0%) and 22.4% may be unable to pay for it. Educational status and awareness of the availability of IVF services were factors that were significantly associated with their overall good perception of IVF services. Also, age, marital status, number of live children, occupation, educational status, awareness, and their overall perception were factors that are significantly associated with their preparedness to utilize IVF services. Conclusion Overall, women's opinions of IVF and their readiness to use them were favorable, and they think it offers hope for their condition since they were well-informed about its forms and that infertility may be a result of several factors, all of which may need IVF services. It does not matter if it is difficult to obtain, expensive, or unavailable. It is recommended that the government collaborates with healthcare providers to investigate ways through the mass media in the drive to clear the misconceptions and improve the public understanding of the IVF procedure towards its utilization, thereby reducing the burden of childlessness and the resulting psychological disorders among couples, this has implications for joyful homes and societal growth.

P-782 The quality and utility of randomised controlled trials in in vitro fertilisation: an analysis of the published literature

Abstract Study question What is the quantity, quality and utility of research on in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) in the last decade? Summary answer Current IVF/ICSI research is of moderate quality, and can be improved by widening inclusion criteria for patient selection, and reporting live-birth as a cumulative measure. What is known already Assisted reproductive technology (ART) is the cornerstone of fertility treatment worldwide for male, female and unexplained infertility. In the last few decades, ART and IVF techniques and protocols have changed considerably, guided by research in the field. Many researchers and journals worldwide continue to publish on ART and IVF yearly, to varying qualities and levels of evidence. Our study aims to analyse the published IVF literature over the last decade and comment on its quality, utility and its feasibility in translating the results into clinical practice. Study design, size, duration We conducted a meta-epidemiological study based on a PubMed search of IVF randomised controlled trials (RCTs) published from 2010 to the present. All articles returned from the initial search were screened by two researchers for relevance, and any conflicts were resolved by a third researcher. We used newly-developed checklists to score usefulness of research, analysing parameters including participant baseline characteristics reported, participant inclusion/exclusion criteria, main study outcomes reported, and whether sample sizes were statistically significant. Participants/materials, setting, methods We selected full text English-language articles, presenting primary data. We included studies comparing either two modalities of IVF, comparing IVF to another ART intervention, or those evaluating the effectiveness of IVF adjunct therapies. Studies focusing on embryo transfer (ET) and ICSI were included, as well as studies focusing on treating complications of IVF such as ovarian hyperstimulation syndrome (OHSS). Studies focusing on intrauterine insemination (IUI), psychosocial outcomes of IVF and lifestyle modifications were excluded. Main results and the role of chance Our initial search returned 5,577 articles. Subsequently, we applied filters for publication date (01/01/2010 to 23/08/2021), Human species, and article types (clinical trials, controlled clinical trials, and randomised clinical trials). The remaining 1,290 articles were manually screened for relevance and adherence to inclusion criteria, leaving 631 RCTs. For this abstract, we analysed the 163 RCTs published in the years 2010, 2015 and 2020, and found that the annual number of publications remained relatively stable (53, 56 and 54, respectively). Regarding participants’ baseline characteristics, 80/163 (49.1%) of studies stated the duration of infertility; only 3/163 (1.8%) of studies stated recurrent miscarriage rates. In terms of study outcomes, 78/163 (47.9%) of studies reported live birth rate (LBR) as an outcome, however only 13/163 (8.0%) of studies reported cumulative LBR, with the remaining studies reporting LBR per first cycle or first transfer. When analysing the participant inclusion, 63/163 (38.7%) of studies had number of cycles as a criterion, with 13/63 (20.6%) of those only including participants in their first cycle. Additionally, 57/163 (35.0%) of studies stated BMI as an inclusion criterion, with 36/57 (63.2%) of those only including participants with BMI of 30 or less. Limitations, reasons for caution Due to practicality and feasibility reasons, we were unable to analyse articles not in English, thus overlooking research published in many parts of the world. Additionally, some parameters we scored are subjective in nature. Moreover, journals have discrepancies in their submission requirements, which may also affect our analysis outcomes. Wider implications of the findings IVF research has many areas for improvement. Of interest are the inclusion criteria for participants: a significant proportion of studies required patients to have a normal BMI, and only included women undergoing their first IVF cycle. A widening of these criteria would increase the applicability of research to clinical practice. Trial registration number not applicable

P-259 Cumulus cells mitochondrial membrane potential as a marker of oocyte competence in assisted reproduction

Abstract Study question Is mitochondrial function of human Cumulus-oophurus Cells (hCCs) an important non-invasive biomarker for oocyte competence and a predictor of in vitro fertilisation (IVF) outcomes? Summary answer Advanced maternal age of women undergoing IVF is characterized by an impairment in cumulus cell mitochondrial function. What is known already The increasing delay of childbearing has led to urgent development of new non-invasive biomarkers of oocyte competency. Aging is a complex process that includes impairment of mitochondrial function and metabolic activity, increase of oxidative stress, alteration in the activity of several cell-signaling systems. A great candidate to monitoring oocyte quality is the hCCs. During folliculogenesis there is an intense bidirectional communication between oocyte and its follicular cells. During oogenesis, CCs supply the oocyte with energy substrates and essential nutrients. Age-related mitochondrial dysfunction in hCCs was described to adversely affect oocyte growth and development. Study design, size, duration The objective of this study was to characterise the mitochondrial membrane potential (Δψm) of hCCs provided from patients undergoing ICSI (intracytoplasmic sperm injection) or oocyte cryopreservation in the Reproductive Medicine Unit of CHUC (Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal) between July and December 2021, to establish a predictive biomarker of oocyte competency and successful IVF outcomes. Participants/materials, setting, methods hCCs, collected from cumulus-oocyte complexes of 53 patients, were incubated with tetramethylrhodamine methyl ester (TMRM - 50 nM), MitoTracker Green (MTG 100 nM) and Hoechst (5µg/mL). Briefly, TMRM, due to its positive charge, accumulates within the mitochondria in a ΔΨm-depend manner, and emits red fluorescence. On the other hand, MTG is green-fluorescent mitochondrial dye which appears to localize in mitochondria regardless of Δψm. Therefore, the red/green fluorescence ratio can be measured as mitochondrial activity outcome. Main results and the role of chance The 53 patients studied were divided in two age groups (27 patients with &amp;lt;35 and 26 with ≥ 35 years old). A decrease in hCCs Δψm of women with advanced maternal age (≥35 y) compared with hCCs recovered from women with &amp;lt;35 was observed (P = 0.001). No differences were found in CCs Δψm in terms of infertility cause, body mass index and social habits (e.g., smoking) (P &amp;gt; 0.05). We also investigated if Δψm can be an effective indicator of clinical outcomes in IVF techniques; however, no correlation was observed between Δψm and oocyte maturation, fertilization, D3 embryo development and blastocyst development rates. Our preliminary data strongly support the idea that advanced reproductive age of women undergoing IVF is accompanied by impaired cumulus cell mitochondrial function. Limitations, reasons for caution More data is needed to be collected to support our preliminary results, such as increasing the sample size, as well as better characterisation of the mitochondrial function in hCCs. Wider implications of the findings This knowledge opens a new window of opportunities in the development of new strategies to mitigate the effects of oocyte aging on (in)fertility, through clinical therapies, such as enhancing the mitochondrial function in hCCs. Trial registration number -

Systemic vascular resistance may influence the outcome of in vitro fertilization

Introduction: The number of pregnancies obtained through in vitro fertilization (IVF) techniques are increasing, and only few studies have investigated hemodynamic variations in women undergoing IVF techniques. The aim of this study was to evaluate the hemodynamic parameters in women undergoing IVF, to assess a possible correlation between hemodynamics and embryo implantation. Methods: 45 normotensive non-obese women, age ≤ 43 years, with idiopathic or tubal infertility, referred to the Reproductive Physiopathology and Andrology Unit, Sandro Pertini Hospital, Rome, during the period 2020/2021, underwent IVF techniques. All women were evaluated with Ultra Sonic Cardiac Output Monitor (USCOM) to detect hemodynamic parameters at two different stages: at the mid-luteal phase, before the beginning of IVF, and at the day of embryo transfer (dET). All demographics and hormonal parameters in both groups were comparable. The hemodynamic parameters were compared between women with a positive β-HCG test vs. those testing negative. Results: 11 out of 45 (24,5%) women obtained positive β-HCG test. All demographics and hormonal parameters were comparable in both groups. Women with a positive β-HCG test showed statistically lower systemic vascular resistance (SVR) at mid-luteal phase (868.61 ± 100.1 vs. 1009 ± 168.4) and dET (818,9 ± 104.5 vs 1038.52 ± 150.82 dynes × s/cm5). Conclusions: Hemodynamic assessment can identify a more favorable pre-pregnancy cardiovascular adaptation. Embryo implantation might be positively influenced by the hemodynamic parameters, e.g. lower SVR, before the beginning of IVF techniques, and during the window of implantation.

Heterologous in vitro fertilization and embryo production for assessment of jaguar (Panthera onca Linnaeus, 1758) frozen-thawed semen in different extenders.

Heterologous in vitro fertilization (IVF) is an important tool for assessing fertility of endangered mammals such as the jaguar, considering difficult access to females for artificial insemination and to obtain homologous oocytes. We aimed to evaluate the fertility of jaguar sperm cryopreserved with different extenders, using domestic cat oocytes to assess the development of hybrid embryos. Semen from four captive jaguars was obtained by electroejaculation. Samples were cryopreserved in powdered coconut water (ACP-117c) or Tris extender containing 20% egg yolk and 6% glycerol. Thawed spermatozoa were resuspended (2.0 × 106 spermatozoa/mL) in IVF medium and co-incubated with cat oocytes matured in vitro for 18 h. Presumptive zygotes were cultured for 7 days. After 48 h, cleavage rate was evaluated, and non-cleaved structures were stained for IVF evaluation. On days 5 and 7, the rate of morula and blastocyst formation was assessed. Data were analyzed using the Fisher exact test (p < 0.05). No difference was observed between ACP-117c and Tris extenders, respectively, for oocytes with 2nd polar body (2/51, 3.9 ± 2.9% vs. 2/56, 3.6 ± 3.1%), pronuclear structures (5/51, 9.8 ± 4.7% vs. 8/56, 14.3 ± 8.0%), and total IVF rates (7/36, 19.4 ± 5.0% vs. 10/37, 27.0 ± 13.8%). All the samples fertilized the oocytes, with 22.9 ± 3.2% (16/70) and 16.7 ± 3.6% (12/72) cleavage of mature oocytes for ACP-117c and Tris extenders, respectively. Morula rates of 4.3 ± 2.3% (3/70) and 5.6 ± 2.2% (4/72) were observed for ACP-117c and Tris, respectively. Only the Tris extender demonstrated blastocyst production (2/12, 16.7 ± 1.5% blastocyst/cleavage). We demonstrated that jaguar ejaculates cryopreserved using ACP-117c and Tris were suitable for IVF techniques, with blastocyst production by ejaculates cryopreserved in Tris. This is a first report of embryos produced in vitro using jaguar sperm and domestic cat oocytes through IVF.

Confrontment and solution to gonadotropin resistance and low oocyte retrieval in in vitro fertilization for type I BPES: a case series with review of literature

BackgroundFOXL2 mutations in human cause Blepharophimosis, ptosis, and epicanthus inversus syndrome (BPES). While type II BPES solely features eyelid abnormality, type I BPES involves not only eyelid but also ovary, leading to primary ovarian insufficiency (POI) and female infertility. Current mainstream reproductive option for type I BPES is embryo or oocyte donation. Attempts on assisted reproductive technology (ART) aiming biological parenthood in this population were sparse and mostly unsuccessful.Case presentationTwo Chinese type I BPES patients with low anti-müllerian hormone (AMH) and elevated follicle stimulating hormone (FSH) presented with primary infertility in their early 30s. Genetic studies confirmed two heterozygous duplication mutations that were never reported previously in East Asian populations. They received in vitro fertilization (IVF) treatment and both exhibited resistance to gonadotropin and difficulty in retrieving oocytes in repeated cycles. Doubled to quadrupled total gonadotropin doses were required to awaken follicular response. Patient 1 delivered a baby girl with the same eyelid phenotype and patient 2 had ongoing live intrauterine pregnancy at the time of manuscript submission.ConclusionsThis is the second reported live birth of biological offspring in type I BPES patients, and first success using IVF techniques. It confirmed that ART is difficult but feasible in type I BPES. It further alerts clinicians and genetic counsellors to type female BPES patients with caution in view of the precious and potentially narrowed reproductive window.

Lars Hamberger: in memory of a Nordic pioneer in IVF.

Professor Emeritus Lars Hamberger, former Chief Editor of Acta Obstetricia et Gynecologica Scandinavica (AOGS), passed away at the age of 82 years on 2 June 2021. Lars Hamberger was an internationally renowned scientist in the field of assisted reproductive technology (ART) and was the initiator and team leader behind the first in vitro fertilization (IVF) baby within the Nordic countries. The birth, in 1982 at Sahlgrenska University Hospital in Gothenburg, occurred only 4 years after the world’s first IVF baby was born. This accomplishment cemented Sweden’s reputation within the international IVF scene as the 5th country in the world to successfully develop the revolutionary technique. Lars Hamberger continued his scientific and clinical work in ART throughout his career, inspiring many young colleagues in Sweden and around the world to pursue careers within this rapidly evolving field. Lars Hamberger studied medicine at the University of Gothenburg and worked at Sahlgrenska University Hospital for most of his career. His doctorate training was in reproductive physiology, specifically ovarian metabolism. His innovative mind led to the development of a micro-diver technique to measure the oxygen consumption of ovarian cells. He became Associate Professor in Physiology in 1968, some months after receiving his PhD. After completing residency training at Sahlgrenska, Hamberger became an Associate Professor in Obstetrics & Gynecology in 1974. He was active in conducting research on ovarian physiology and closely followed the international work carried out to develop a technique that would fertilize the human oocyte outside the body. The successful creation of this IVF technique was largely due to the scientific efforts of Robert Edwards in Cambridge. In 1978, the Cambridge group reported the birth of Louise Brown. Soon after, Hamberger formed a team of clinicians and scientists to set up IVF in close contact with the Cambridge team, proving successful in 1982 with their first live birth at Sahlgrenska. In 2010, Robert Edwards was awarded the Nobel prize in physiology/medicine for the scientific development of IVF. Lars Hamberger was appointed Professor of Obstetrics & Gynecology at Karolinska Institute in 1983 but returned to the University of Gothenburg the following year as a professor. IVF was soon established as a clinical treatment at Sahlgrenska and Hamberger created a large adjoining research laboratory, “Spec-lab”, for translational research in ART. This center attracted scientists from all over the world to work in developing IVF and ART even further. One major IVF invention from the Sahlgrenska center was the forming of transvaginal ultrasound-guided oocyte retrieval in 1987. With this technique it was possible to avoid laparoscopy surgery when obtaining oocytes. Another major invention in ART was intracytoplasmic sperm injection (ICSI), developed by the Brussels group in 1992. Lars Hamberger and his team quickly learned the technique and Sahlgrenska became the second center in the world with a live birth after ICSI. Several major research findings in ART came from the Sahlgrenska center, including the establishment of elective single embryo transfer as a routine, long-term follow-up of IVF children, and uterus transplantation. Alongside all the science in the laboratory, Hamberger also made space for the world-renowned photographer Lennart Nilsson, who created the book “A Child is Born”, with Lars Hamberger as co-author. The book has been translated into 18 languages. Lars Hamberger was not only a scientist and clinician but also an entrepreneur within the business field. He was co-founder of the second private IVF clinic in Sweden, the stem-cell company Cellartis, as well as Vitro Life, now a major global company within the area of Culture media and products for assisted reproduction. Lars Hamberger supervised several PhD students within the Departments of Physiology and Obstetrics & Gynecology and hosted a great number of international visiting scientists. The famous Spec-lab became an international hub for advanced research in ART, greatly aided by Hamberger’s scientific generosity and hospitality. Lars Hamberger became honorary member of the European Society of Human Reproduction (ESHRE) in 2011 and was the Chief Editor of AOGS from 1999 to 2000. On a personal level, Lars had many interests but was particularly passionate about the arts. Lars himself was a talented painter and as a young man dreamed of a future as a painter, despite the disapproval of his father, Professor Carl-Axel Hamberger. However, Hamberger pursued his painting in parallel with his scientific career and had several art exhibitions after retirement. Lars Hamberger will be remembered with warmth and gratitude for being instrumental in the establishment of ART in the Nordic countries and being essential to the development of AOGS.