Oocyte Donation Research Articles

Anti-Müllerian hormone as a predictor of oocyte yield following controlled ovarian stimulation in the rhesus macaque.

Anti-Müllerian hormone (AMH) is widely used in the clinic as a biomarker for ovarian reserve and to predict ovarian response to gonadotropin stimulation. Patients with higher AMH levels tend to yield more oocytes and have better outcomes from assisted reproductive technology (ART) procedures. The goal of this study is to determine if AMH can be used to predict the outcome of controlled ovarian stimulation in rhesus macaques, which are commonly used in biomedical research, to refine animal use while maximizing oocyte yield. We hypothesized that pre-stimulation AMH values can be used to predict oocyte yield and quality. Regularly cycling adult macaques underwent controlled ovarian stimulation and baseline (pre-stimulation) plasma AMH levels were determined using an AMH-specific enzyme-linked immunoassay. Oocytes were collected by laparoscopic or ultrasound-guided aspiration, then counted and evaluated for quality and stage of meiosis. Sperm from established fertile males were used to inseminate the oocytes in vitro with fertilization success checked 14 - 16 hours later. Females were grouped by oocyte yield: low ≤ 17; mid = 18 - 41; high ≥ 42. We found that high and mid yielders had significantly higher AMH than low yielders (p<0.0001) and the percent of mature oocytes was greater in the high and mid yielders. There were no significant differences in oocyte quality or ova fertilization rate. These data suggest that AMH is a useful measure for controlled ovarian stimulation success in rhesus macaques and can be used to identify suitable animals for oocyte donation before entering them into a stimulation protocol.

Global DNA methylation level in spermatozoa is not associated with ICSI fertilization outcome and embryo quality in donor oocyte programme

Epigenetic mechanisms, including DNA methylation and histone modifications, govern chromatin arrangement in sperm, enhancing motility and safeguarding DNA integrity for accurate epigenetic inheritance. Abnormal methylation is linked to poor sperm quality and fertility issues, underscoring the need to study sperm DNA methylation and its impact on sperm function and embryo development in assisted reproductive technology. In this study, processed spermatozoa from 75 normozoospermic and 15 abnormal ejaculates were examined for sperm global DNA methylation levels using a colourimetric absorbance method. Although semen characteristics were poor in abnormal ejaculates, no significant correlation was found between sperm global DNA methylation levels and sperm characteristics in either normozoospermic or abnormal cohorts. However, mean global DNA methylation levels were significantly lowered in abnormal sperm samples compared to normozoospermic samples (p < 0.05). Furthermore, injecting spermatozoa from these patients (N = 50) into donor oocytes did not show a significant relationship between sperm global DNA methylation and embryo developmental competence. These findings highlight the limitation of sperm global DNA methylation as a biomarker for embryo development and quality.

Phthalates are detected in the follicular fluid of adolescents and oocyte donors with associated changes in the cumulus cell transcriptome

ObjectiveTo investigate follicular fluid (FF) phthalate levels in adolescents undergoing fertility preservation compared to oocyte donors and explore its association with ovarian reserve and cumulus cell gene expression. DesignRetrospective study, and molecular analysis of cumulus cells (CCs) and FF SubjectsAdolescents (n= 20, 16.7 ± 0.6 years old) undergoing fertility preservation and oocyte donors (n= 24, 26.2 ± 0.4 years old). Main Outcome MeasuresPatient demographics, ovarian stimulation, and oocyte retrieval outcomes were analyzed for each group. FF levels of 9 phthalate metabolites were assessed individually and as molar sums representative of common compounds (all phthalates: ƩPhthalates; di(2-ethylhexyl) phthalate (DEHP) associated phthalate metabolites: ƩDEHP), exposure sources (plastics: ƩPlastic; personal care products: ƩPCP), and modes of action (anti-androgenic: ƩAA) and compared between the two groups. The transcriptome of CC associated with mature oocytes was compared between adolescents and oocyte donors using bulk RNA-sequencing. ResultsFollicular fluid ƩPlastic and ƩPCP levels were significantly higher in adolescents compared to oocyte donors (p<0.05). Follicular fluid ƩDEHP, ƩPlastic, ƩPCP, ƩAA, and ƩPhthalates levels were positively associated with antral follicle count (AFC) (p<0.05) in oocyte donors when adjusted for age, BMI, and race/ethnicity. RNA-seq analysis revealed 248 differentially expressed genes (DEGs) in cumulus cells of adolescents within the top quartile (n=4) of FF ƩPhthalates levels compared to the adolescents within the bottom half (n=9). Genes enriched in pathways involved in cell motility and development were significantly downregulated. ConclusionAdolescents undergoing fertility preservation cycles demonstrate higher levels of phthalate metabolites in their follicular fluid compared to oocyte donors. Phthalate metabolite levels in FF are associated with higher AFC levels in oocyte donors.

Air pollution exposure in vitrified oocyte donors and male recipient partners in relation to fertilization and embryo quality.

Studies on air pollution and outcomes of in vitro fertilization (IVF) have focused on couples undergoing autologous IVF, in which it is challenging to disentangle maternal and paternal exposures during gametogenesis. We sought to evaluate the independent associations between air pollution exposure during oogenesis and spermatogenesis on fertilization and embryo quality in non-identified donor oocyte IVF cycles. Our study included 500 oocyte donors and 915 male recipient partners who contributed 1,095 oocyte thaw cycles (2008-2019). Daily ambient air pollutant exposure was estimated using spatio-temporal models based on residential address and averaged over folliculogenesis (i.e., three months prior to initiation of controlled ovarian stimulation), controlled ovarian stimulation, and spermatogenesis (i.e., 72days prior to oocyte thaw). We used multivariable generalized estimating equations to estimate the adjusted odds ratios (aOR) and 95% confidence intervals (CI) for an interquartile range increase in pollutant exposure in relation to the proportion of oocytes surviving thaw, oocytes fertilized, and usable embryos. Oocyte donors with higher exposure to organic carbon (OC) (aOR=0.86 95%CI 0.79,0.94) and particulate matter<10µm (aOR=0.69 95%CI 0.54,0.90) during folliculogenesis had a lower proportion of oocytes surviving thaw. During ovarian stimulation, higher particulate matter<2.5µm (aOR=0.78 95%CI 0.66, 0.91), nitrate (aOR=0.83 95% CI 0.69,0.99), and OC (aOR=0.86 95% CI 0.80,0.93) exposure was associated with a lower proportion of surviving oocytes while nitrogen dioxide (aOR=1.11 95%CI 1.00,1.23) and ozone (aOR=1.19 95%CI 1.04,1.37) exposure was associated with a higher proportion of fertilized oocytes and usable embryos. Elemental carbon (aOR=0.93 95%CI 0.87,1.00) and OC (aOR=0.95 95%CI 0.90,1.00) exposure during spermatogenesis was associated with a slightly lower proportion of usable embryos. On the day of oocyte thaw, higher ambient OC at the IVF clinic was associated with lower oocyte survival and higher ozone was associated with lower fertilization. Both maternal and paternal air pollution exposures during gametogenesis have independent, largely detrimental, effects on early embryological outcomes.

Recipients' age, fresh embryo and blastocyst-stage embryo transfer as favorable factors in a transnational oocyte donation program.

In Italy, egg donation is being allowed again following the ruling by the Constitutional Court N. 162/2014. Nevertheless, the number of voluntary donors is insignificant, probably because no reimbursement is provided. Therefore, the great majority of egg donation cycles are performed by using imported cryopreserved oocytes from foreign countries. However, recent evidence has questioned the overall efficacy of this strategy in comparison with the shipment of frozen sperm and vitrified embryos. Retrospective cohort study, conducted at ANDROS Day Surgery Clinic Palermo, Italy in the period 2016-2022, with the objective of evaluating the efficiency of each batch of vitrified oocytes, in terms of completed cycles. The cryopreserved donated oocytes came from a single Spanish clinic (Next Fertility, Valencia, Spain). The primary outcome was the cumulative live birth rate (CLBR). The secondary outcomes were the cumulative ongoing pregnancy rate (COPR), the cumulative clinical pregnancy rate (CCPR) and the miscarriage rate. The CLBR totaled 35.1%, the COPR 36.8%, the CCPR 49.2% and the miscarriage rate 24.7%. In the multivariate analysis, the probability of success was significantly higher in younger recipients, in fresh embryo transfer and in blastocyst-stage embryo transfer. Our "transnational" donation program with vitrified oocytes has good success rates. The main focus of our analysis is the negative effect of advancing recipient age on success rates, outlining a possible, important role of age-related uterine factor on implantation mechanisms. This aspect should be emphasized during the counselling with couples much more than is routinely done.

Proteomic Analysis of Follicular Fluid in Polycystic Ovary Syndrome: Insights into Protein Composition and Metabolic Pathway Alterations.

This study explores the proteomic composition of follicular fluid (FF) from women undergoing oocyte retrieval for in vitro fertilisation (IVF), with a focus on the effects of polycystic ovary syndrome (PCOS). FF samples were collected from 74 patients, including 34 with PCOS and 40 oocyte donors. Proteomic profiling using machine learning identified significant differences in protein abundance between the PCOS and control groups. Of the 484 quantified proteins, 20 showed significantly altered levels in the PCOS group. Functional annotation and pathway enrichment analysis pointed to the involvement of protease inhibitors and immune-related proteins in the pathophysiology of PCOS, suggesting that inflammation and immune dysregulation may play a key role. Additionally, HDL assembly was identified as a significant pathway, with apolipoprotein-AI (APOA1) and alpha-2-macroglobulin (A2M) as the major proteins involved. Notably, myosin light polypeptide 6 was the most downregulated protein, showing the highest absolute fold change, and may serve as a novel independent biomarker for PCOS.

Non-genetic risk factors for preeclampsia: an updated umbrella review

Abstract Background Preeclampsia (PE) is a common pregnancy disorder with complex mechanisms, and multiple risk factors may contribute to its development. Our objective is to update our literature search from our previous umbrella review, summarizing recent evidence on non-genetic risk factors associated with PE and assessing their credibility to identify those supported by robust epidemiological evidence. Methods We searched PubMed, EMBASE, Scopus, and ISI Web of Science up to June 2023. For each meta-analysis, we computed summary effect estimates, 95% confidence intervals, 95% prediction intervals, and assessed heterogeneity using I², while also examining small-study effects and excess significance bias. Quality assessment was conducted using the AMSTAR-2 tool. Results 89 meta-analyses were identified, providing data on 219 associations. Of these, 151 (69%) showed statistically significant findings at P &amp;lt; 0.05, with 52 (24%) remaining significant at P &amp;lt; 10-6. Large or very large heterogeneity was observed in 121 (55%) associations. Evidence for small-study effects and excess significance bias was found in 27 (12%) and 29 (13%) associations, respectively. Only 16 (7%) presented convincing evidence: Polycystic Ovary Syndrome (PCOS), PCOS (adjusted), Obstructive Sleep Apnea (OSA), Obesity (adjusted), Chronic Kidney Disease (CKD), Pregnancy Intrahepatic Cholestasis, SARS-CoV-2 infection (adjusted), Oocyte donation (OD) vs. In vitro fertilization (IVF), Fresh embryo transfer vs. spontaneous conception (SC) (singleton), Frozen embryo transfer vs. SC, OD vs. SC (singleton), IVF-OD vs. IVF-AO, IVF-OD fresh ET vs. IVF-AO fresh ET, NC frozen embryo transfer vs. AC frozen embryo transfer, Fetal sex (term), and Smoking. Conclusions Maternal health conditions like PCOS, along with reproductive interventions such as OD and IVF, as well as lifestyle factors like obesity and smoking, showed convincing associations with PE. Key messages • Maternal health conditions, reproductive interventions, and lifestyle factors showed compelling evidence of association with preeclampsia. • Our findings emphasize the multifactorial nature of preeclampsia and suggest potential avenues for further research and clinical intervention.

Stereological study of organelle distribution in human mature oocytes

The ultrastructure of human oocytes has been described only qualitatively. To offer a precise organelle spatial distribution and organelle volume during the main maturation stages, we previously conducted stereological studies on prophase-I (GV) and metaphase-I (MI) oocytes, and here we present results on metaphase-II (MII) oocytes. Five donor oocytes from different donors were processed for transmission electron microscopy, and quantification of organelle distribution was performed using point-counting stereology. Statistical tests compared the means of the relative volumes occupied by organelles among oocyte regions. The most abundant organelles were elements of the smooth endoplasmic reticulum (SER), such as SER small vesicles, SER medium vesicles, SER large vesicles and SER isolated tubules, along with mitochondria, followed by SER tubular aggregates, cortical vesicles and lysosomes. Significant differences between oocyte regions were found for lysosomes, cortical vesicles and SER large vesicles. Comparisons of MII oocytes to previous findings in GV and MI oocytes evidenced specific patterns of organelle distribution and relative volumes. This final evaluation thus enables to track organelle spatial reorganization across oocyte stages, which, in addition to gathered knowledge, may be useful to assist in improvements of stimulation protocols, in-vitro maturation media and cryopreservation techniques.

An exploratory study of perceptions and utilization of genetic information in the intended parent experience of oocyte donor selection.

Utilization of oocyte donation has become an increasingly common practice in assisted reproductive technology (ART). Since the introduction of larger carrier screening (CS) panels and extended family medical histories (EFMH), studies have not examined how this information factors into the oocyte donor selection process. This exploratory, qualitative study provides further insight into what role, if any, donors' available genetic information (e.g., larger CS panels and EFMH) plays in selecting an oocyte donor. An online screening survey was distributed to individuals who have undergone or are currently in the process of selecting an oocyte donor through the RESOLVE network and Mayo Clinic's Reproductive Endocrinology and Infertility clinic. From 13 survey respondents, six oocyte recipients subsequently participated in semi-structured telephone interviews and discussed their experiences as oocyte recipients including their perceptions of donors' available genetic information and process in choosing an oocyte donor. Genetic information was seen as valuable and reassuring for participants, particularly EFMH, but did not play a significant role in the selection process for these participants. Supplemental emergent themes provide context on the psychosocial complexities of the oocyte recipient experience and possible explanations for why genetic information is not a decisional priority. Participants indicated genetic information was not extensively discussed or fully explained by providers. Results demonstrate how genetic counselors can be more involved in the pre-selection process to discuss the utility and limitations of genetic information, as well as address psychosocial issues common to the oocyte recipient experience.

Progestin-primed ovarian stimulation outcomes in in-vitro fertilization (IVF) – A systematic review of the literature

Background: Progestin-primed ovarian stimulation (PPOS) stimulates ovaries to block the premature surge of luteinizing hormone (LH) by using micronized progesterone or a progestin during the follicular phase instead of the conventional gonadotropin-releasing hormone (GnRH) analogues or GnRH antagonists downregulating LH to obtain multi-follicle engagement. Current work aims to assess the influence of progestogen treatment on ovarian stimulation and the ability to control LH surge, its efficacy and suitability in retrieving oocytes, without affecting the embryo quality and its benefit among infertile women long-term outcomes on children compared to standard stimulation protocols. Materials and Methods: The literature review used the randomized control trials published in the Pubmed database from January 2015 to April 2021. To generate the citation list, the following keywords were used: ‘progestin-primed ovarian stimulation’, ‘PPOS’, ‘micronized progesterone’, ‘medroxyprogesterone’, and/or ‘dydrogesterone’. The selected articles analyzed the cohort, intervention, and scheme of the progestin-primed ovarian stimulation protocol in controlled ovarian stimulation (COS) for in-vitro fertilization (IVF)/intra cytoplasmic sperm injection (ICSI) used in Assisted Reproductive Technologies (ART). Results: Overall we concluded that PPOS for IVF/ICSI in ART results in a higher number of obtained embryos, lower incidence of OHSS, equal duration of stimulation, number of retrieved oocytes, and number of MII oocytes. It is also suggested that long-term safety in children shows no significant difference between the study and control groups. Conclusions: Despite the outcomes of progestin stimulation cycles among all cohorts, we concluded that poor ovarian responders, patients with PCOS, women of advanced age and oocyte donors benefit the most from using PPOS.

Diagnosing a genetic disease in a donor-conceived child: case report and discussion of the ethical, legal, and practical issues.

We describe the unexpected challenges pediatricians may experience when children conceived with assisted reproduction are diagnosed with a rare genetic condition. A local case series triggered a dialogue between many stakeholders with varied expertise. Indeed, diagnosing a genetic disease in a child conceived by embryo, egg, or sperm donation is becoming more common now that genetic testing and in vitro fertilization (IVF) are readily accessible. However, how and whether to share that information with other stakeholders in the gamete donation process has not been fully explored, and the clinical responsibilities of the treating clinician remain ill-defined. This work centers on a patient with a confirmed diagnosis of X-linked nephrogenic diabetes insipidus. Ultimately, we found the same AVPR2 mutation in two children from two families conceived by egg donation. It led to multigenerational cascade diagnoses in the family of the shared, anonymous egg donor. First, we review current genetic testing practices in gamete donation and third-party reproduction. Then, we provide an overview of relevant genomic, ethical, legal, and psychosocial considerations for sharing relevant genomic information. Finally, and to maximize the best interests of genetic relatives, we discuss how a pediatrician can play a role in the early disclosure of relevant clinical information to all stakeholders in the gamete donation process, starting with the fertility clinic. While these clinical cases are cast in a Canadian context, we submit that its important lessons generally apply to medical systems of nearly all developed countries, broadly defined.

Fertility knowledge, childbearing intentions and attitudes towards social egg freezing: A study of female students of a private university in Kumasi, Ghana.

As women increasingly delay childbearing for educational and career pursuits, understanding the age-related decline in ovarian reserve becomes crucial to avoid infertility. Egg freezing offers a solution, extending the window for achieving pregnancy with a biological child. However, awareness and acceptance of age-related fertility decline and elective egg freezing vary across countries and social groups. This descriptive cross-sectional study investigates fertility awareness, childbearing intentions, and attitudes toward elective egg freezing among 138 undergraduate female students at a private university in Kumasi, Ghana. Data were collected via a Google App-based questionnaire. The majority of respondents were below 28 years old (76.3%), Christians (79.26%), and single (57. 78%). Many respondents overestimated their knowledge of age-related fertility decline, with almost half (48.89%) incorrectly believing it occurs after age 40. Primary sources of fertility education included health professionals, the Internet, and television. While most (62.96%) considered childbearing necessary, they would not have a child without a partner. Acceptance of donor egg in vitro fertilisation (IVF) varied based on circumstances, and a significant portion was open to donating their eggs (41.48%). Awareness of elective egg freezing was low, but nearly half (48.89%) expressed interest in it, primarily to retain the option for a biological child. The cost of elective egg freezing was a significant deterrent. The study revealed a significant lack of awareness regarding age-related fertility decline and elective egg freezing among female university students in Kumasi. Most prefer having a biological child only with a partner. While few were open to donating their eggs or accepting the same to achieve a pregnancy, interest in elective egg freezing was notable, albeit hindered by cost. The study recommends enhanced education by health professionals using social media and television to address these gaps.

The effect of male factors on embryo morphokinetics: a retrospective analysis of 2726 blastocysts.

Male infertility may influence fertilization rates, embryo morphology, and implantation rates in in vitro fertilization (IVF) cycles. Oocyte competence plays a major role in embryo development, but there is a limited understanding of the connection between sperm quality, embryo development, and morphokinetic parameters using donor oocytes. The study evaluated if sperm quality may influence the morphokinetic parameters in IVF cycles. A retrospective multicentric observational cohort study included 747 ICSI cycles using donor oocytes with fresh or frozen sperm. Embryos were cultured in time-lapse incubators until the blastocyst stage. The population was divided into three groups according to sperm concentration, as control group (> 16 mill/mL), severe oligospermia (0-5 mill/mL), and moderate oligospermia group (5-16 mill/mL). Morphokinetic analysis showed no difference in the time from the 2-cell to 6-cell stage of embryo development. A significant difference was observed on day 3 of embryo development, specifically at the 7-cell stage (t7), severe oligospermia 53.37 ± 9.81, moderate oligospermia 56.95 ± 9.78, and control 55.1 ± 8.85h post-insemination (hpi) (p = 0.024), and 8-cell stage (t8), severe oligospermia 55.41 ± 10.83, moderate oligospermia 61.86 ± 12.38 hpi (p < 0.001), and control 58.61 ± 11.33. Accordingly, the synchrony of the four cleavages going from 4 to 8 cells (s3) was found statistically different among the groups in the severe oligospermia 8.05 ± 9.99, moderate oligospermia 11.66 ± 11.04 hpi, and control 8.55 ± 8.58 (p = 0.009). Morphokinetic time ranges were obtained for t6, t7, t8, and s3 in order to identify the good-quality blastocysts. Poor sperm quality is associated with alterations in morphokinetic parameters on day 3 in IVF cycles with donor oocytes, underlining the important role of spermatozoa during embryo development.

The Business of Egg Transactions and Need for Improved Regulation of the Fertility Industry in Canada

In Canada, ethical and legal debates around egg donation have largely focused on the question of whether egg providers should be paid, since payment for egg donation is prohibited through the federal Assisted Human Reproduction Act, 2004. One of the generally overlooked issues, however, is the potential for fertility clinics and agencies to mistreat egg providers. There is little legal oversight of the practices of fertility clinics and agencies in Canada, and scholars have raised concerns about the potential for mistreatment of egg providers. This article examines the medical care of egg providers in Canada by presenting data from qualitative interviews with 14 egg providers. While some egg providers felt well cared for, others were left feeling like a means to an end — they felt that insufficient information was provided for consent, there was an inability to communicate with the clinic, and they were physically mistreated. I argue that not only do egg providers’ experiences run contrary to the CMA Code of Ethics, but they also run contrary to laws, regulations, and guidelines that are intended to ensure that people who undergo fertility treatments are not harmed.

Female-age-dependent changes in the lipid fingerprint of the mammalian oocytes.

Can oocyte functionality be assessed by observing changes in their intracytoplasmic lipid droplets (LDs) profiles? Lipid profile changes can reliably be detected in human oocytes; lipid changes are linked with maternal age and impaired developmental competence in a mouse model. In all cellular components, lipid damage is the earliest manifestation of oxidative stress (OS), which leads to a cascade of negative consequences for organelles and DNA. Lipid damage is marked by the accumulation of LDs. We hypothesized that impaired oocyte functionality resulting from aging and associated OS could be assessed by changes in LDs profile, hereafter called lipid fingerprint (LF). To investigate if it is possible to detect differences in oocyte LF, we subjected human GV-stage oocytes to spectroscopic examinations. For this, a total of 48 oocytes derived from 26 young healthy women (under 33 years of age) with no history of infertility, enrolled in an oocyte donation program, were analyzed. Furthermore, 30 GV human oocytes from 12 women were analyzed by transmission electron microscopy (TEM). To evaluate the effect of oocytes' lipid profile changes on embryo development, a total of 52 C57BL/6 wild-type mice and 125 Gnpat+/- mice were also used. Human oocytes were assessed by label-free cell imaging via coherent anti-Stokes Raman spectroscopy (CARS). Further confirmation of LF changes was conducted using spontaneous Raman followed by Fourier transform infrared (FTIR) spectroscopies and TEM. Additionally, to evaluate whether LF changes are associated with developmental competence, mouse oocytes and blastocysts were evaluated using TEM and the lipid dyes BODIPY and Nile Red. Mouse embryonic exosomes were evaluated using flow cytometry, FTIR and FT-Raman spectroscopies. Here we demonstrated progressive changes in the LF of oocytes associated with the woman's age consisting of increased LDs size, area, and number. LF variations in oocytes were detectable also within individual donors. This finding makes LF assessment a promising tool to grade oocytes of the same patient, based on their quality. We next demonstrated age-associated changes in oocytes reflected by lipid peroxidation and composition changes; the accumulation of carotenoids; and alterations of structural properties of lipid bilayers. Finally, using a mouse model, we showed that LF changes in oocytes are negatively associated with the secretion of embryonic exosomes prior to implantation. Deficient exosome secretion disrupts communication between the embryo and the uterus and thus may explain recurrent implantation failures in advanced-age patients. Due to differences in lipid content between different species' oocytes, the developmental impact of lipid oxidation and consequent LF changes may differ across mammalian oocytes. Our findings open the possibility to develop an innovative tool for oocyte assessment and highlight likely functional connections between oocyte LDs and embryonic exosome secretion. By recognizing the role of oocyte LF in shaping the embryo's ability to implant, our original work points to future directions of research relevant to developmental biology and reproductive medicine. This research was funded by National Science Centre of Poland, Grants: 2021/41/B/NZ3/03507 and 2019/35/B/NZ4/03547 (to G.E.P.); 2022/44/C/NZ4/00076 (to M.F.H.) and 2019/35/N/NZ3/03213 (to Ł.G.). M.F.H. is a National Agency for Academic Exchange (NAWA) fellow (GA ULM/2019/1/00097/U/00001). K.F. is a Diamond Grant fellow (Ministry of Education and Science GA 0175/DIA/2019/28). The open-access publication of this article was funded by the Priority Research Area BioS under the program "Excellence Initiative - Research University" at the Jagiellonian University in Krakow. The authors declare no competing interest. N/A.

First-trimester uterine artery pulsatility index and preeclampsia risk in pregnancies after artificial frozen embryo transfer: analysis of over 27,000 pregnancies

Accumulating evidence indicates that pregnancies after Artificial Cycle Frozen Embryo Transfer are associated with an increased risk of preeclampsia. Uterine Artery Doppler, along with maternal factors and serum biomarkers, is a crucial biomarker for first-trimester preeclampsia screening, aiding in identifying "high-risk" patients. Guidelines strongly recommend administering aspirin (150mg/day) in these women, owing to robust evidence demonstrating a 62% reduction in the incidence of preeclampsia. Although previous studies suggested lower Uterine Artery Pulsatility Index after Frozen Embryo Transfer, no previous studies explored the impact of the type of endometrial preparation in Uterine Artery Doppler or its influence on estimating 1st-trimester preeclampsia risk. The study aims to evaluate the possible impact of endometrial preparation for frozen embryo transfer on the Uterine Artery Pulsatility Index during the first-trimester preeclampsia screening. This is a retrospective single-center study including 27289 singleton pregnancies (naturally conceived or after assisted reproductive treatment) who underwent the 1st-trimester ultrasound screening at our University Hospital between January 2010 and May 2023. Overall, 27289 pregnancies were included: 23410 naturally conceived and 3879 following assisted reproductive technologies including 391 after ovulation induction and intrauterine insemination, 888 in-vitro fertilization and fresh embryo transfer, and 2600 natural or artificial frozen embryo transfer cycles. An Analysis of covariance (ANCOVA) was conducted to assess if there is an association between the UtAPI value and the mood of conception, adjusting for confounding factors (age, weight, smoking, and oocyte donation). Overall, pregnancies after artificial frozen embryo transfer demonstrated significantly lower 1st-trimester Uterine Artery Pulsatility Index as compared with all other modes of conception in a multivariable regression analysis adjusted for age, weight, smoking, and oocyte donation. The percent difference was 22.6 [CI95%: 20,6; 24,5] compared to naturally conceived pregnancy, 24.5 [CI95%:20,7; 28,1] to Ovulation Induction or intrauterine insemination, 24.8 [CI95%: [22,9; 27,6] to fresh Embryo Transfer and 21.7 [CI95%: [17,6; 25,5] compared to Natural Cycle Frozen Embryo Transfer. When calculating the risk for initiating preventive aspirin administration, the number of patients with increased risk (>1/100) who initiated prophylactic aspirin was significantly lower in the artificial cycle frozen embryo transfer group (7.8% vs 16.0% in natural cycle p<0.001 vs. 11.0% in Fresh embryo transfer p=0.01 vs. 10.5% in ovulation induction or intrauterine insemination p=0.14 vs. 9,3% in naturally conceived pregnancy p=0.03). Surprisingly although significantly fewer patients were considered at high risk for preeclampsia in the Artificial Cycle Frozen Embryo Transfer group, analysis of the actual incidence of preeclampsia demonstrated three times higher preeclampsia incidence in Artificial Cycle group 5.3% (122/2284) as compared with naturally conceived 1.4 % (321/23410), Ovulation Induction and intrauterine insemination 1.3 % (5/391) or Natural Cycle pregnancies 1.6 % (5/316) and more than two times higher when compared to fresh Embryo Transfer pregnancies 2.3 % (20/888), p<0.001. Pregnancies following frozen embryo transfer in artificial cycle are associated with significantly lower Uterine Artery Pulsatility Index during 1st-trimester preeclampsia screening. This results in a significantly lower number of patients being classified as high-risk for developing preeclampsia, despite accumulating evidence that artificial cycles are linked to an increased risk of preeclampsia. Therefore, the first-trimester preeclampsia risk algorithm should be adjusted to accurately assess risk for those undergoing Artificial Cycle Frozen Embryo Transfer, to prevent the under-treatment of patients who are at very high risk of developing preeclampsia and may benefit from prophylactic aspirin.

Effects of herbal medicine on ovulation in premature ovarian insufficiency patients during in vitro fertilization

Introduction: Premature ovarian insufficiency (POI) is a condition in which menopause occurs before the age of 40 and is known to be a relatively common disease that occurs in 1% of all women. The only way for patients diagnosed with POI to become pregnant is through oocyte donation, in which fertilized ovums are transplanted by receiving oocytes from donors.4 Can POI patients increase their chances of normal ovulation? POI patients were more likely to observe and collect follicles larger than the average 15 mm diameter when herbal medicine was administered. Methods: This study is a retrospective cohort study conducted on POI patients undergoing in vitro fertilization (IVF) who visited our hospital from 2020 to 2022. We would like to report the response rate to gonadotropin-releasing hormone (GnRH) during IVF in the hyperovulation induction low response group after herbal medicine treatment. Statistical analysis was performed using IBM’s SPSS Statistics to verify significance with a P-value&lt;0.05. Results: In recovering ovarian POI patient's ovarian function, herbal medicine treatment increased the response rate to GnRH (the number of oocytes collected) during IVF, follicle-stimulating hormone (FSH) and cortisol levels decreased, and estradiol (E2) and luteinizing hormone (LH) levels increased. Conclusions: It was confirmed that herbal medicine treatment can increase the probability of normal ovulation in patients with idiopathic POI with high cortisol levels. Until now, it has been known that ovarian function cannot be recovered once irreversibly decreased, but by increasing the probability of ovulation through herbal medicine treatment, it is significant that it may be possible to suppress ovarian aging or preserve fertility.

Oocyte donation pregnancies with high fetal-maternal immunogenetic dissimilarity show alterations in the maternal peripheral immunoregulatory response

Oocyte donation (OD) pregnancies result in increased fetal-maternal immunogenetic dissimilarity due to paternal and donor-derived genes. Higher fetal-maternal HLA mismatches are correlated with preeclampsia. Therefore, this study explored the maternal immune response, focusing on regulatory T cells (Tregs), during low versus high allogeneic pregnancies, and healthy versus preeclamptic OD pregnancies. Ten healthy and five preeclamptic OD pregnancies were included. Maternal peripheral blood was collected at different stages of pregnancy. Fetal-maternal HLA mismatches were determined, and immunophenotyping of peripheral blood mononuclear cells was conducted using a 22-colour spectral flow cytometry panel. Cytokines and hormones were detected in maternal plasma using ELISA and Luminex assays. The findings show similarities, but also distinct differences between low and high allogeneic healthy OD pregnancies. Early high allogeneic OD pregnancy showed reduction in Tregs, and CD8+ T cells, alongside lower percentage of effector/memory Tregs expressing PD-1 and Helios. Additionally, high allogeneic OD pregnancies showed increased IL-6 and progesterone in the first trimester. These variations suggest a different mode of immune regulation in early high allogeneic OD pregnancies, possibly to maintain healthy pregnancy. Further comparative analyses revealed reduced CD45RO+CTLA-4+ Tregs and increased latent TGF-β1 and -β2 levels in early preeclamptic compared to healthy OD pregnancy. Late-stage preeclamptic OD pregnancies exhibited higher frequencies of CD45RO+TIGIT+ Tregs and higher levels of TNFα, indicating both a regulatory and pro-inflammatory environment. Overall, this study sheds light on the course of various immunoregulatory key players in OD pregnancy, and expands knowledge on maternal tolerance in this particular type of pregnancy.

External validation of a fully automated evaluation tool: a retrospective analysis on 68,471 scored embryos.

To externally validate a fully automated embryo classification in in vitro fertilization (IVF) treatments. Retrospective cohort study SUBJECTS: A total of 6,434 patients undergoing 7,352 IVF treatments contributed 70,456 embryos. Embryos were evaluated by conventional morphology and retrospectively scored using a fully automated deep learning-based algorithm across conventional IVF, oocyte donation, and PGT-A cycles. The primary outcomes were implantation and live birth including odds ratios (ORs) from generalized estimating equation (GEE) models. Secondary outcomes were embryo morphology, euploidy and miscarriage. Exploratory outcomes included comparison between conventional methodology and artificial intelligence (AI) algorithm with areas under the ROC curves (AUCs), agreement degree between AI and embryologists, Cohen's Kappa coefficient and relative risk (RR). Implantation and live birth rates increased as the automatic embryo score rose. The GEE model, controlling for confounders, showed the automatic score was associated with an OR of 1.31 (95%CI[1.25-1.36]) for implantation in treatments using oocytes from patients, and an OR of 1.17 (95%CI[1.14-1.20]) in the oocyte donation program, with no significant association in PGT-A treatments. For live birth, the ORs were 1.27 (95%CI[1.21-1.33]) for patients, 1.16 (95%CI[1.13-1.19]) for donors, and 1.05 (95%CI[1-1.10]) for PGT-A cycles. The average score was higher in embryos with better morphology, in euploid embryos compared to aneuploid embryos, and in embryos that resulted in a full-term pregnancy compared to those that miscarried. Concordance between the highest-scoring embryo and the embryo with the best conventional morphology was 71.4%(95%CI[67.7%-75.0%]) in treatments with patient oocytes and 61.0%(95%CI[58.6%-63.4%]) in the oocyte donation program. Overall, the Cohen's Kappa coefficient was 0.63. The automatic embryo score showed similar AUCs to conventional morphology, although implantation was higher when the transferred embryo matched the highest-scoring embryo from each cohort (57.36% vs. 49.98%). RR indicated a 1.14-fold increase in implantation likelihood when the top-ranked embryo was transferred. Fully automated embryo scoring effectively ranked embryos based on their potential for implantation and live birth. The performance of the conventional methodology was comparable to that of the artificial intelligence-based technology; however, better clinical outcomes were observed when the highest-scoring embryo in the cohort was transferred.

Testing an artificial intelligence algorithm to predict fetal heartbeat of vitrified-warmed blastocysts from a single image: predictive ability in different settings.

Could an artificial intelligence (AI) algorithm predict fetal heartbeat from images of vitrified-warmed embryos? Applying AI to vitrified-warmed blastocysts may help predict which ones will result in implantation failure early enough to thaw another. The application of AI in the field of embryology has already proven effective in assessing the quality of fresh embryos. Therefore, it could also be useful to predict the outcome of frozen embryo transfers, some of which do not recover their pre-vitrification volume, collapse, or degenerate after warming without prior evidence. This retrospective cohort study included 1109 embryos from 792 patients. Of these, 568 were vitrified blastocysts cultured in time-lapse systems in the period between warming and transfer, from February 2022 to July 2023. The other 541 were fresh-transferred blastocysts serving as controls. Four types of time-lapse images were collected: last frame of development of 541 fresh-transferred blastocysts (FTi), last frame of 467 blastocysts to be vitrified (PVi), first frame post-warming of 568 vitrified embryos (PW1i), and last frame post-warming of 568 vitrified embryos (PW2i). After providing the images to the AI algorithm, the returned scores were compared with the conventional morphology and fetal heartbeat outcomes of the transferred embryos (n = 1098). The contribution of the AI score to fetal heartbeat was analyzed by multivariate logistic regression in different patient populations, and the predictive ability of the models was measured by calculating the area under the receiver-operating characteristic curve (ROC-AUC). Fetal heartbeat rate was related to AI score from FTi (P < 0.001), PW1i (P < 0.05), and PW2i (P < 0.001) images. The contribution of AI score to fetal heartbeat was significant in the oocyte donation program for PW2i (odds ratio (OR)=1.13; 95% CI [1.04-1.23]; P < 0.01), and in cycles with autologous oocytes for PW1i (OR = 1.18; 95% CI [1.01-1.38]; P < 0.05) and PW2i (OR = 1.15; 95% CI [1.02-1.30]; P < 0.05), but was not significantly associated with fetal heartbeat in genetically analyzed embryos. AI scores from the four groups of images varied according to morphological category (P < 0.001). The PW2i score differed in collapsed, non-re-expanded, or non-viable embryos compared to normal/viable embryos (P < 0.001). The predictability of the AI score was optimal at a post-warming incubation time of 3.3-4 h (AUC = 0.673). The algorithm was designed to assess fresh embryos prior to vitrification, but not thawed ones, so this study should be considered an external trial. The application of predictive software in the management of frozen embryo transfers may be a useful tool for embryologists, reducing the cancellation rates of cycles in which the blastocyst does not recover from vitrification. Specifically, the algorithm tested in this research could be used to evaluate thawed embryos both in clinics with time-lapse systems and in those with conventional incubators only, as just a single photo is required. This study was supported by the Regional Ministry of Innovation, Universities, Science and Digital Society of the Valencian Community (CIACIF/2021/019) and by Instituto de Salud Carlos III (PI21/00283), and co-funded by European Union (ERDF, 'A way to make Europe'). M.M. received personal fees in the last 5 years as honoraria for lectures from Merck, Vitrolife, MSD, Ferring, AIVF, Theramex, Gedeon Richter, Genea Biomedx, and Life Whisperer. There are no other competing interests. N/A.