Age-related Fertility Decline Research Articles

Gallic acid supplementation partially ameliorates reproductive aging in rooster breeders by improving semen quality, sperm kinetics, hormones, and antioxidant status

Aging leads to decreased fertility in roosters, which is likely due to increased oxidative stress. This study evaluated the antioxidant effects of gallic acid (GA) supplementation on sperm quality and fertility of aged roosters. This study evaluated whether gallic acid (GA) supplementation can mitigate age-related fertility decline. Roosters were randomly assigned to: control, 100 mg/kg GA, or 200 mg/kg GA. Semen parameters, sperm kinetics, hormone levels, fertility rate, and hatchability were assessed. GA increased semen concentration, membrane integrity and viability while decreasing defects versus control (P<0.01). Testosterone was higher in GA groups (P<0.01) without affecting gonadotropins. Furthermore, 200 mg/kg GA optimized motility, velocity, linearity, and beat cross frequency versus control and 100 mg/kg GA (P<0.01). Fertility and hatchability were higher in both GA groups. In conclusion, GA supplementation in aged roosters improves sperm quality, antioxidant status, testosterone, and fertility outcomes, likely by mitigating oxidative stress. The 200 mg/kg dose elicited optimal effects on motion parameters.

“You feel like you’re fairly disadvantaged with an advert over your head saying ‘in final years of reproduction’”: social egg freezing, dating and the (unequal) politics of reproductive ageing

Recent decades have seen an increasing gap occurring between the ‘desired’ and ‘actual’ family size of middle-class and professional women. This situation of ‘unrealised fertility’ and ‘incomplete families’ carries implications at a population, but also couple and individual level. This paper explores how middle-class professional women make decisions about partnering and parenthood and how these are shaped by a contemporary neoliberal feminist discourse which articulates the possibility of ‘having it all’ through engagement in careful life planning, appropriate self-investment, and by drawing on new technologies of reproductive biomedicine. Informed by semi-structured interviews with UK and US women conducted at two different points in time, it explores how they approach and experience the process of relationship formation in the face of age-related fertility decline. It also examines how the use of social egg freezing shape their romantic and family building expectations but also their interactions with (potential) partners. In doing so, it explores how gendered cultural dating scripts and unequal gender power relations shape the formation and progression of intimate relationships in a manner which can disempower women as they age. It therefore questions whether egg freezing may be the ‘great equaliser’ that some have hoped.

Cryptochrome 1 regulates ovarian granulosa cell senescence through NCOA4-mediated ferritinophagy

Age-associated decreases in follicle number and oocyte quality result in a decline in female fertility, which is associated with increased infertility. Granulosa cells play a major role in oocyte development and maturation both in vivo and in vitro. However, it is unclear whether a reduction in cryptochrome 1 (Cry1) expression contributes to granulosa cell senescence, and further exploration is needed to understand the underlying mechanisms. In this study, we investigated the role of Cry1, a core component of the molecular circadian clock, in the regulation of senescence in ovarian granulosa cells. Western blotting and qRT-PCR showed that Cry1 expression was downregulated in aged human ovarian granulosa cells and was correlated with age and anti-Müllerian hormone (AMH) levels. RNA-seq analysis suggested that ferritinophagy was increased after Cry1 knockdown in KGN cells. MDA, iron, and reactive oxygen species (ROS) assays were used to detect cellular ferritinophagy levels. Ferroptosis inhibitors, iron chelators, autophagy inhibitors, and nuclear receptor coactivator 4 (NCOA4) knockdown alleviated KGN cell senescence induced by Cry1 knockdown. Immunofluorescence, immunoprecipitation, and ubiquitination assays indicated that Cry1 affected NCOA4 ubiquitination and degradation through HERC2, thereby affecting NCOA4-mediated ferritinophagy and causing granulosa cell senescence. KL201, a Cry1 stabilizer, enhanced ovarian function in naturally aged mice by reducing ferritinophagy. Our study reveals the potential mechanisms of action of Cry1 during ovarian aging and provides new insights for the clinical treatment of age-related fertility decline.

Mechanisms of endometrial aging: lessons from natural conceptions and assisted reproductive technology cycles.

Until recently, the study of age-related decline in fertility has focused primarily on the ovary; depletion of the finite pool of oocytes and increases in meiotic errors leading to oocyte aneuploidy are well-established mechanisms by which fertility declines with advancing age. Comparatively little is known about the impact of age on endometrial function. The endometrium is a complex tissue comprised of many cell types, including epithelial, stromal, vascular, immune and stem cells. The capacity of this tissue for rapid, cyclic regeneration is unique to this tissue, undergoing repeated cycles of growth and shedding (in the absence of an embryo) in response to ovarian hormones. Furthermore, the endometrium has been shown to be capable of supporting pregnancies beyond the established boundaries of the reproductive lifespan. Despite its longevity, molecular studies have established age-related changes in individual cell populations within the endometrium. Human clinical studies have attempted to isolate the effect of aging on the endometrium by analyzing pregnancies conceived with euploid, high quality embryos. In this review, we explore the existing literature on endometrial aging and its impact on pregnancy outcomes. We begin with an overview of the principles of endometrial physiology and function. We then explore the mechanisms behind endometrial aging in its individual cellular compartments. Finally, we highlight lessons about endometrial aging gleaned from rodent and human clinical studies and propose opportunities for future study to better understand the contribution of the endometrium to age-related decline in fertility.

Oocytes on ice: Exploring the advancements in elective egg freezing for women

Introduction: Female fecundity decreases significantly after the age of 32, and rapidly so after age 37. There is no treatment to prevent this decline. Furthermore, globally, women are getting married later and the age at which they have their first child is increasing. As of July 2023, elective egg freezing (EEF) or oocyte cryopreservation (OC) for age-related fertility decline, commenced in Singapore. With medical advancements in OC, EEF is no longer considered experimental. The aim of this review is to examine the existing literature around EEF with regard to reproductive outcomes and its safety, to better guide clinicians in counselling young single women. Method: Published studies were examined to increase understanding on optimal age for EEF, ideal number of oocytes for a live birth, recommended OC protocols, cryopreservation techniques affecting thaw survival or fertilisation, oocyte storage and pregnancy risks. Results: Models predict that EEF should be performed at age &lt;37 years and to achieve a 70% chance of live birth, women would need 14, 15 and 26 mature oocytes at ages 30–34, 35–37 and &gt;38 years, respectively. An antagonist stimulation protocol with an agonist trigger would minimise ovarian hyperstimulation syndrome and duration of stimulation without affecting outcomes. Oocyte vitrification in comparison to slow freezing increases thaw survival, fertilisation and clinical pregnancy rates. No increased risks exist for the woman, future pregnancy or child when compared with conventional IVF. Conclusion: EEF is a viable option for single women desiring fertility preservation. Financial costs are significant, but returns are worthwhile if oocytes are utilised.

FROM PATHOPHYSIOLOGY TO POTENTIAL INTERVENTIONS: INVESTIGATING THE INTRICATE DYNAMICS OF POLYCYSTIC OVARY SYNDROME, AGING, AND FERTILITY

Polycystic ovary syndrome (PCOS) and age-related decline in fertility are complex issues affecting women’s reproductive health. This review aims to explore the intricate dynamics between PCOS, aging, and fertility, and examine potential interventions for improved clinical outcomes. It delves into the pathophysiology of PCOS, highlighting hormonal imbalances, metabolic disturbances, and the impact on ovarian function and follicular development. Concurrently, age-related changes in female fertility are discussed, including ovarian reserve decline, hormonal shifts, and alterations in egg quality and uterine environment. Emphasizing the overlapping mechanisms, shared pathways are elucidated such as hormonal dysregulation, insulin resistance, hyperandrogenism, and inflammation. Additionally, the clinical implications of diagnosing and managing PCOS in aging women are addressed, as well as fertility preservation options and assisted reproductive techniques. Furthermore, potential interventions and therapeutic strategies are explored, encompassing lifestyle modifications, pharmacological interventions, and emerging technologies. By comprehensively reviewing the current evidence and research gaps, this article provides valuable insights into the interplay between PCOS, aging, and fertility, paving the way for optimized clinical approaches and future advancements.

Single-cell transcriptomic atlas of goat ovarian aging

BackgroundThe ovaries are one of the first organs that undergo degenerative changes earlier in the aging process, and ovarian aging is shown by a decrease in the number and quality of oocytes. However, little is known about the molecular mechanisms of female age-related fertility decline in different types of ovarian cells during aging, especially in goats. Therefore, the aim of this study was to reveal the mechanisms driving ovarian aging in goats at single-cell resolution.ResultsFor the first time, we surveyed the single-cell transcriptomic landscape of over 27,000 ovarian cells from newborn, young and aging goats, and identified nine ovarian cell types with distinct gene-expression signatures. Functional enrichment analysis showed that ovarian cell types were involved in their own unique biological processes, such as Wnt beta-catenin signalling was enriched in germ cells, whereas ovarian steroidogenesis was enriched in granulosa cells (GCs). Further analysis showed that ovarian aging was linked to GCs-specific changes in the antioxidant system, oxidative phosphorylation, and apoptosis. Subsequently, we identified a series of dynamic genes, such as AMH, CRABP2, THBS1 and TIMP1, which determined the fate of GCs. Additionally, FOXO1, SOX4, and HIF1A were identified as significant regulons that instructed the differentiation of GCs in a distinct manner during ovarian aging.ConclusionsThis study revealed a comprehensive aging-associated transcriptomic atlas characterizing the cell type-specific mechanisms during ovarian aging at the single-cell level and offers new diagnostic biomarkers and potential therapeutic targets for age-related goat ovarian diseases.

#391 : Oocyte Cryopreservation — A Five Year Follow Up Study on its Utilization and Outcome

Background &amp; Aim: Oocyte cryopreservation is a boon for women undergoing Assisted reproductive technology (ART). Apart from cancer patients it is recently promoted as a mode of fertility insurance to overcome the age-related decline in fertility, endometriosis, primary ovarian insufficiency, etc. The objective of the study was to evaluate the clinical applications of oocyte freezing and its efficacy in terms of pregnancy outcome. Method: Retrospective observational study of all patients undergoing oocyte cryopreservation from 2016 to 2021 in Lilavati Hospital &amp; Research Centre, Mumbai, India. Result: 221 patients undergoing oocyte cryopreservation were included. Oocyte cryopreservation was done for different indications like azoospermia (50.2%, most common), planned oocyte freezing (41.17%), fertility preservation (1.36%), endometriosis (0.45%), ejaculatory dysfunction (1.8%), husband with active hepatitis B infection (3.62%) and unavailability of husband on the day of egg retrieval (3.62%). A total of 1715 mature oocytes were frozen. 41.17% women were normo-responder, 38.91% were poor responder and 19.90% were hyper-responder. The oocytes of 59 patients (return rate-26.7%) have been thawed and 162 women (73.3%) remain in storage. Out of 508 oocytes frozen for these 59 patients utilizing their oocytes, around 474 oocytes were recovered post-warming (recovery rate 93.3%). A total of 59.3% patients created embryos with a partner, and 40.6% used donor sperm. The mean fertilization rate was 65.8% of surviving oocytes. Of 59 patients, 44 (74.5%) planned a fresh embryo transfer, out of which 20 patients tested positive for pregnancy (pregnancy rate 45.45%). Conclusion(s): We report the outcomes of patients opting for oocyte cryopreservation, which is useful for patient counselling. Limited evidence is available in terms of utilization rate, post warming survival of oocytes, fertilization and pregnancy rate. Further studies over long period of time with larger cohorts are needed.

Effect of Lifestyle in Female Infertility: A Review Based Study

Abstract: Female infertility is a multifaceted issue with numerous contributing factors. While medical conditions and genetics play a significant role, recent research has shed light on the compelling influence of lifestyle choices. This research paper explores the intricate relationship between lifestyle and female infertility, delving into the impact of dietary habits, physical activity, substance use, and stress management on a woman’s reproductive health. Through an extensive review of existing literature, this paper synthesizes findings that emphasize the critical role of lifestyle factors in fertility. It highlights the potential consequences of diets rich in processed foods, excessive alcohol consumption, smoking, and sedentary behavior. Conversely, it underscores the benefits of a balanced diet, moderate exercise, and stress reduction in promoting fertility and overall well-being. Furthermore, this paper examines the interplay between lifestyle and age, recognizing that the age-related decline in fertility can be influenced and, to some extent, mitigated by healthy living choices. It also explores the impact of environmental factors and socioeconomic disparities on female infertility. This research contributes to a comprehensive understanding of the modifiable aspects of female infertility and their implications for healthcare practices, public health policies, and individual decisionmaking. It advocates for a holistic approach to reproductive health, where lifestyle choices are not only recognized as integral but also as actionable pathways toward improved fertility outcomes. This research underscores the significance of lifestyle in the realm of female infertility and the potential for proactive measures to enhance reproductive health. It sets the stage for a future where individuals and healthcare providers can harness the power of lifestyle choices to support fertility, destigmatize infertility, and prioritize overall well-being.

Exploring the knowledge, attitudes, and perceptions of women of reproductive age towards fertility and elective oocyte cryopreservation for age-related fertility decline in the UK: a cross-sectional survey.

What are the knowledge, perceptions and attitudes towards fertility and elective oocyte cryopreservation (OC) for age-related fertility decline (ARFD) in women in the UK? Awareness of OC for ARFD has reportedly improved compared to studies carried out almost a decade ago, but inconsistencies in knowledge remain regarding the rate of miscarriage amongst specific age groups, the financial costs and optimal age to undergo OC for ARFD. The age of first-time motherhood has increased amongst western societies, with many women of reproductive age underestimating the impact of age on fertility. Further understanding of women's awareness of their fertility, the options available to preserve it and the barriers for seeking treatment earlier are required in order to prevent the risk of involuntary childlessness. A hyperlink to a cross-sectional survey was posted on social media (Instagram) between 25 February 2021 and 11 March 2021. Women from the general population aged 18-50 years were invited to complete the survey. In total, 5482 women fulfilled the inclusion criteria and completed the survey. The mean age of participants was 35.0 years (SD 10.25; range 16-52). Three quarters (74.1%; n = 4055) disagreed or strongly disagreed they felt well informed regarding the options available to preserve their fertility, in case of a health-related problem or ARFD. The majority overestimated the risk of miscarriage in women aged ≥30 years old, with 14.5% correctly answering 20%, but underestimated the risks in women ≥40, as 20.1% correctly answered 40-50%. Three quarters (73.2%; n = 4007) reported an awareness of OC for ARFD and 65.8% (n = 3605) reported that they would consider undergoing the procedure. The number of women who considered OC for ARFD across age groups were as follows: 18-25 (8.3%; n = 300), 26-30 (35.8%; n = 1289), 31-35 (45.9%; n = 1654), 36-40 (9.6%; n = 347), 41-45 (0.3%; n = 13), and 46-50 (0.1%; n = 2). The majority of women (81.3%; n = 4443) underestimated the cost of a single cycle of OC for ARFD (<£5000). Furthermore, 10.4% (n = 566) believed a single cycle would be adequate enough to retrieve sufficient oocytes for cryopreservation. Approximately 11.0% (n = 599) believed OC for ARFD may pose significant health risks and affect future fertility. Less than half agreed or strongly agreed that the lack of awareness regarding OC for ARFD has impacted the likelihood of pursuing this method of fertility preservation further (41.4%; n = 2259). Results from cross-sectional studies are limited as interpretations made are merely associations and not of causal relationships. The online nature of participant recruitment is subject to selection bias, considering women with access to social media are often from higher socioeconomic and education backgrounds, thus limiting generalizability of the findings. Further education regarding the financial costs and optimal age to undergo elective OC to increase the chances of successful livebirth are required. Clinicians should encourage earlier fertility counselling to ensure that OC is deemed a preventative measure of ARFD, rather than an ultimate recourse to saving declining fertility. No funding was required for this article. There are no conflicts of interests to declare. N/A.

Human ovarian aging is characterized by oxidative damage and mitochondrial dysfunction.

Are human ovarian aging and the age-related female fertility decline caused by oxidative stress and mitochondrial dysfunction in oocytes? We found oxidative damage in oocytes of advanced maternal age, even at the primordial follicle stage, and confirmed mitochondrial dysfunction in such oocytes, which likely resulted in the use of alternative energy sources. Signs of reactive oxygen species-induced damage and mitochondrial dysfunction have been observed in maturing follicles, and even in early stages of embryogenesis. However, although recent evidence indicates that also primordial follicles have metabolically active mitochondria, it is still often assumed that these follicles avoid oxidative phosphorylation to prevent oxidative damage in dictyate arrested oocytes. Data on the influence of ovarian aging on oocyte metabolism and mitochondrial function are still limited. A set of 39 formalin-fixed and paraffin-embedded ovarian tissue biopsies were divided into different age groups and used for immunofluorescence analysis of oxidative phosphorylation activity and oxidative damage to proteins, lipids, and DNA. Additionally, 150 immature oocytes (90 germinal vesicle oocytes and 60 metaphase I oocytes) and 15 cumulus cell samples were divided into different age groups and used for targeted metabolomics and lipidomics analysis. Ovarian tissues used for immunofluorescence microscopy were collected through PALGA, the nationwide network, and registry of histo- and cytopathology in The Netherlands. Comprehensive metabolomics and lipidomics were performed by liquid-liquid extraction and full-scan mass spectrometry, using oocytes and cumulus cells of women undergoing ICSI treatment based on male or tubal factor infertility, or fertility preservation for non-medical reasons. Immunofluorescence imaging on human ovarian tissue indicated oxidative damage by protein and lipid (per)oxidation already at the primordial follicle stage. Metabolomics and lipidomics analysis of oocytes and cumulus cells in advanced maternal-age groups demonstrated a shift in the glutathione-to-oxiglutathione ratio and depletion of phospholipids. Age-related changes in polar metabolites suggested a decrease in mitochondrial function, as demonstrated by NAD+, purine, and pyrimidine depletion, while glycolysis substrates and glutamine accumulated, with age. Oocytes from women of advanced maternal age appeared to use alternative energy sources like glycolysis and the adenosine salvage pathway, and possibly ATP which showed increased production in cumulus cells. The immature oocytes used in this study were all subjected to ovarian stimulation with high doses of follicle-stimulating hormones, which might have concealed some age-related differences. Further studies on how to improve mitochondrial function, or lower oxidative damage, in oocytes from women of advanced maternal age, for instance by supplementation of NAD+ precursors to promote mitochondrial biogenesis, are warranted. In addition, supplementing the embryo medium of advanced maternal-age embryos with such compounds could be a treatment option worth exploring. The study was funded by the Amsterdam UMC. The authors declare to have no competing interests. N/A.

Rethinking advanced motherhood: a new ethical narrative

The aim of the study is to rethink the ethics of advanced motherhood. In the literature, delayed childbearing is usually discussed in the context of reproductive justice, and in relationship to ethical issues associated with the use and risk of assisted reproductive technologies. We aim to go beyond these more “traditional” ways in which reproductive ethics is framed by revisiting ethics itself through the lens of the figure of the so-called “older” mother. For this purpose, we start by exploring some of the deep seated socio-cultural discourses in the context of procreation: ageism, ableism and the widespread bias towards geneticism and pronatalism. Afterwards, we provide a critical overview of the key arguments against or in support of advanced motherhood. We then briefly discuss how entrenchment by both sides has produced an impasse in the debate on the ethics of advanced motherhood and proceed by arguing that it is fundamental to bring about a change in this narrative. For this purpose, we will revisit the feminist usage of the concept of vulnerability which will allow us both to criticize culturally prescribed norms about motherhood and to address the painful reality of age-related fertility decline. In the last section, we argue that instead of defining “older” motherhood as an ethical problem, we should problematize the fact that female reproductive ageing is an understudied and ill-sourced topic. We believe that allocating resources to research to better understand female reproductive ageing is not only ethically permissible, but might even be ethically desirable.

P-399 Oocyte vitrification does not impair clinical outcomes in women of advanced maternal age. An analysis of 1268 cycles using fresh and vitrified oocytes

Abstract Study question Does oocyte vitrification impair clinical outcomes in women of advanced maternal age (AMA, &amp;gt;35 years old)? Summary answer Up to the age of 39 years, clinical outcomes were comparable amongst cycles using fresh and vitrified oocytes. What is known already Primarily recommended for young women, oocyte vitrification is the most efficient method for elective fertility preservation. Nevertheless, various social and economic circumstances often compel women to undergo fertility preservation when already at an AMA. Recent studies in mouse suggest that AMA increases the vulnerability of oocytes to molecular and subcellular damage during vitrification, ultimately compromising embryo viability and implantation potential. However, the effects of oocyte vitrification on human preimplantation development and clinical outcomes in women of AMA currently remain unknown. Study design, size, duration This is a retrospective cohort single-center study. We compared outcomes of 1268 patients undergoing their first fresh ICSI cycle using autologous fresh (n = 1087) or vitrified oocytes (n = 181), performed between January 2019 and October 2022. All vitrified oocytes were obtained following elective fertility preservation for age-related fertility decline. Outcomes were stratified according to maternal age at oocyte pick-up: control group (≤35) and AMA groups (36-37, 38-39 and ≥40 years). Participants/materials, setting, methods We compared outcomes between fresh and vitrified oocytes across matching maternal age groups. These included fertilization rates, proportion of viable embryos on day 3, clinical pregnancy and live birth rates. Univariate and multivariate analyses (2-contrast linear and logistic regression) were used to establish correlations. Analyses were adjusting for sperm origin (partner or donor), day of transfer (3 or 5), number of embryos transferred and endometrial age. All p values &amp;lt;0.05 were considered significant. Main results and the role of chance Sperm donation and homologous cycles were equally distributed among the study groups (45% sperm donor and 55% male partner across the entire cohort). In our cohort, blastocyst transfer was performed less frequently in older women, 23.8% for women ≥40 years compared to 45.6% for women ≤35 years (p &amp;lt; 0.0001). When comparing oocyte status across matching maternal age groups, fertilization rates were ∼10% lower when vitrified oocytes were used (p &amp;lt; 0.05). Our adjusted analysis confirmed this negative impact of oocyte vitrification on fertilization rates. Oocyte vitrification also affected ongoing cleavage, as the proportion of viable embryos on day 3 was significantly lower across all groups when vitrified oocytes were used (p &amp;lt; 0.05). Nevertheless, this effect did not translate to clinical pregnancy rates, which were comparable across all maternal age groups regardless of oocyte status. As expected, live birth rate decreased steadily with advancing maternal age (from 38.5% in women ≤35 to 15.0% in women ≥40). While our adjusted analysis showed a negative effect of oocyte vitrification on live birth rates for patients ≥40 (OR: 0.60, 95%CI: 0.18-2.06), we observed no such effect in AMA patients ≤39 years. Limitations, reasons for caution The main limitation of this study is its retrospective design. The low number of women ≥40 undergoing fertility presentation (n = 39) warrants careful interpretation. Our results cannot be generalized to all patient populations and indications for fertility preservation. Wider implications of the findings Clinical outcomes were comparable between fresh and vitrified oocytes up to the age of 39 years. While oocyte vitrification before 35 years old remains the most established and effective method for preventing age-related fertility decline, our findings offer a valuable clinical resource for counselling AMA patients considering elective fertility preservation. Trial registration number Not applicable

P-441 Frequency of sexual intercourse and fecundability among women trying to conceive in Japan

Abstract Study question To what extent is the frequency of sexual intercourse associated with fecundability? Summary answer Frequency of sexual intercourse was the most influential factor in achieving pregnancy but was low among women trying to conceive in Japan. What is known already The Japanese National Fertility Survey showed that 39% of married couples had experienced some fertility problems. A recent national survey revealed that 47% of married couples were sexless. The effect of intercourse frequency on time-to-pregnancy (TTP) has not been well investigated in Japan despite the high prevalence of fertility problems and sexless marriages. Study design, size, duration We used a social research panel in Japan to conduct an internet-based preconception cohort study of pregnancy planners who were not undergoing fertility treatment. In total, 3,796 women were enrolled in February 2021 and followed up for six months. We used data from 2,371 (62%) participants who discontinued contraception. Participants/materials, setting, methods Participants were married women between the ages of 25 and 39 who planned to conceive within a year but were not pregnant at the time of enrolment. Those receiving fertility treatment or trying to conceive for more than six months were excluded from the recruitment. Participants reported frequency and timing of intercourse along with sociodemographic and lifestyle factors. TTP was measured in months, and discrete-time Cox regression was used to estimate fecundability odds ratios (FORs). Main results and the role of chance Of the 2,371 women, 31% had intercourse “less than once in a few months” (hereafter, the Lowest Frequency), 54% had intercourse “several times per month” (hereafter, the Middle Frequency), and 15% engaged in intercourse “several times or more per week” (hereafter, the Highest Frequency). Higher frequency groups were younger, were married for a shorter period, and were less likely to have kids (all Ps for trend&amp;lt;0.05). Adjusted FORs of the Middle and the Highest Frequency groups were 2.20 (95% confidence interval [CI]: 1.56—3.10) and 3.41 (95% CI:2.29—5.08), respectively, resulting in a shorter TTP compared to the Lowest Frequency group. Covariates showing significant associations with fecundability were timed intercourse of fertile window (adjusted FOR=1.76, 95% CI: 1.37—2.27), folic acid and/or multivitamin use (adjusted FOR=1.44, 95%CI: 1.12—1.84), and participant age (adjusted FOR=0.96, 95% CI: 0.92—0.99), whereas the other variables, such as partner age, sociodemographic factors, health indicators (e.g., body mass index, cycle regularity), or lifestyles (e.g., smoking, physical activity), did not show significant associations. Limitations, reasons for caution Intention to conceive could be a confounder, although we used timed intercourse as a proxy measure. Misclassification may have occurred due to self-reported information. Wider implications of the findings Occasional intercourse among pregnancy planners might be linked to the high prevalence of infertility in Japan through a direct reduction in fecundability before the age-related fertility decline. Informing couples about the frequency of intercourse and TTP might help them achieve pregnancy earlier. Trial registration number NA

O-081 A 10-year follow-up of reproductive outcomes in women returning after elective oocyte cryopreservation

Abstract Study question What are the reproductive treatment outcomes in women who underwent elective oocyte cryopreservation (EOC) returning to our clinic with a desire for a child? Summary answer Whether to warm oocytes or to first use fresh oocytes for ART depends on age upon returning, but both strategies resulted in favourable reproductive outcomes. What is known already Most affluent countries have observed a trend towards postponement of childbearing, and EOC is increasingly used in an attempt to extend the reproductive lifespan and mitigate age-related fertility decline. Although most follow-up studies after EOC have focused on women who requested oocyte warming, a substantial proportion of women who do not conceive naturally eventually embark for a fertility treatment without using their cryopreserved oocytes. Reports on reproductive outcomes in past EOC users are scarce, and the lack of reproductive treatment algorithms in this group of women hampers efficiency of clinical practice. Study design, size, duration This retrospective observational single-center study includes 843 women who had elective oocyte vitrification for non-medical reasons between 2009 and 2019, and describes the outcomes of the diverse reproductive treatment strategies that were performed in these women when they returned to the same clinic because of a desire for pregnancy until May 2022. Participants/materials, setting, methods Patient characteristics and data of ovarian stimulation of EOC cycles were analyzed, as well as data related to ovarian stimulation and laboratory data of ART in women who pursued fertility treatment with and/or without using their cryopreserved oocytes. The primary outcome was the live birth rate (LBR) per patient after oocyte warming and after ART using fresh oocytes. Secondary outcomes were laboratory outcomes and LBR per embryo transfer. Main results and the role of chance A total of 1353 EOC cycles (1.6 ± 0.9 per patient) were performed. Mean age was 36.5 ± 2.8 years. At the time of EOC 174 (20,6%) women had a partner. On average, 13.7 ± 9.2 mature oocytes were cryopreserved. After 39.9 ± 23.4 months, 231 (27.4%) women returned to the clinic. Upon returning, mean age was 40.4 ± 3.1 years, mean AMH 2.3 ± 2.0 ng/mL, and 150/231 (64,9%) patients had a partner. As a primary approach, 110/231 (47.6 %) past EOC women embarked on oocyte warming, 50/231 (21.6%) had intrauterine insemination, and 71/231 (30.7%) had ART using fresh oocytes. Cumulative LBR (CLBR) was 45.9 % (106/231) and miscarriage rate (MR) was 30.7% (51/166) in the entire cohort. A subset of 90/231 (39.0%) patients exclusively had oocyte warming, at 41.6 ± 3.0 years, with 10.0 ± 5.2 oocytes warmed per patient. 52/231 (22.5%) patients exclusively had ART using fresh oocytes, at a mean age of 39.0 ± 2.8 years, with 9.9 ± 7.4 mature oocytes retrieved per patient. CLBR was 37/90 (41.1%) in the oocyte warming-only group and 25/52 (48.1%) in the OS-only group. MR/transfer was 25.0% and 29.3% in the oocyte warming-only group and the OS-only group, respectively. Limitations, reasons for caution Both the small sample size and the retrospective design are limitations of this study. The decision to embark on a specific reproductive treatment strategy was based on patient preference, after counseling of treatment options. This precludes direct comparison of the efficacy of reproductive treatment options in past EOC users. Wider implications of the findings Reporting of clinical outcomes of women who underwent EOC and returned to the clinic to embark on divergent reproductive treatment strategies is mandatory to establish guidelines for best clinical practice in this growing patient population. Trial registration number Not applicable

P-801 Human umbilical cord perivascular cells (HUCPVC) reduce ovarian fibrosis and improve pregnancy rates in a mouse model of natural ovarian aging

Abstract Study question Does repeated intravenous administration of human umbilical cord perivascular cells (HUCPVC) during the period of ovarian aging prevent age-related fertility decline in a mouse model? Summary answer Repeated administration of first trimester and term HUCPVC improved pregnancy rates and reduced ovarian fibrosis in a mouse model of natural ovarian aging. What is known already Mesenchymal stromal cells (MSC), such as first trimester (FTM) and term human umbilical cord perivascular cells (HUCPVC), may be good cell candidates to mitigate side-effects of oncotherapy, including infertility, and are also gaining interest in ovarian rejuvenation strategies. The intraovarian or systemic administration of various sources of mesenchymal stromal cells in animal models of advanced reproductive age have been previously reported to improve ovarian reserve, follicle activation and function, via multiple proposed mechanisms. However, mating studies have not previously been performed to demonstrate fertility preservation. In addition, the optimal dose regimen and timing of treatment has not been established. Study design, size, duration Pre-clinical randomized controlled study in a mouse model of natural ovarian aging from 6 months to 12 months, including a negative control group (vehicle-treated), a cell control group (fibroblast-treated) and 3 experimental groups (HUCPVC, 2 sources and 2 dose regimens; n = 10-15 per group). All parameters were compared with young control female mice (6-8 weeks, 6 months). Cell injections and all assessments were blinded. Participants/materials, setting, methods 6-month-old ICR mice randomized into 4 groups received 6 monthly tail vein injections of 1x106 HUCPVC (FTM or term), fibroblast cells or HBSS (vehicle control). A 5th group received a single injection of FTM HUCPVC at 11M (n = 5). Pregnancy and litter size data were collected after breeding trials. The total number of follicles per ovary and picrosirius red (PSR) staining were quantified. Serum anti-Mullerian hormone (AMH) and C-reactive protein (CRP) were analyzed by ELISA. Main results and the role of chance No differences in mortality rates (P = 0.2) and animal weights were observed. The weight gain in FTM and term HUCPVC treated animals was significantly greater than in the other groups, over the duration of the study (P = 0.04). Animals that received 6 injections of FTM and term HUCPVC showed significantly higher pregnancy rates at 12M (100%, 80%) (P = 0.007; P = 0.048, respectively), when compared to animals that received HBSS (40%) and were similar to pregnancy rates of 6-8weeks (100%) and 6M (77%) groups. Fibroblast-treated and 1x FTM HUCPVC-treated groups showed 60% pregnancy rates. Age-associated declines in litter sizes and ovarian reserve indicators were observed as expected, but not rescued by cell treatments, except for levels of AMH which were increased in the group that received 1x FTM HUCPVC injections when compared to HBSS controls (P = 0.04). Age-associated increases in ovarian stroma fibrosis were observed with aging in the control groups (P &amp;lt; 0.0001), and were significantly reduced in 6x FTM HUPCVC, 6x term HUCPVC and 1x FTM HUCPVC groups, when compared to HBSS and fibroblast controls (P &amp;lt; 0.0001; P = 0.01 and P &amp;lt; 0.0001, respectively). An age-associated increase in CRP (P = 0.01) was significantly decreased in 6x FTM HUCPVC-treated groups, when compared to HBSS (P = 0.04). Limitations, reasons for caution Pregnancy rate (the main outcome measured in this study) may confound some of the secondary outcomes analyzed (CRP levels). Use of ovarian tissue for histological analysis and limited serum samples precluded further molecular analyses to assess potential mechanisms for the effects of HUCPVC. Wider implications of the findings FTM HUCPVC have anti-inflammatory and anti-fibrotic effects, and represent a promising source of MSC to prevent fertility decline in women of advanced reproductive age. Trial registration number Not Applicable

Oocyte Cryopreservation for Medical and Planned Indications: A Practical Guide and Overview.

Oocyte cryopreservation (OC) is the process in which ovarian follicles are stimulated, the follicular fluid is retrieved, and mature oocytes are isolated and vitrified. Since the first successful pregnancy utilizing previously cryopreserved oocytes in 1986, OC has become increasingly utilized as an option for future biologic children in patients facing gonadotoxic therapies, such as for the treatment of cancer. Planned OC, also termed elective OC, is growing in popularity as a means to circumvent age-related fertility decline. In this narrative review, we describe both medically indicated and planned OC, focusing on the physiology of ovarian follicular loss, OC technique and risks, timing of when OC should be performed, associated financial considerations, and outcomes.

Altered microRNA expression profiles of human spermatozoa in normal fertile men of different ages.

MicroRNAs (miRNAs) are mediators of the aging process. The purpose of this work was to analyze the miRNA expression profiles of spermatozoa from men of different ages with normal fertility. Twenty-seven donors were divided into three groups by age (Group A, n = 8, age: 20-30 years; Group B, n = 10, age: 31-40 years; and Group C, n = 9, age: 41-55 years) for high-throughput sequencing analysis. Samples from 65 individuals (22, 22, and 21 in Groups A, B, and C, respectively) were used for validation by quantitative real-time polymerase chain reaction (qRT-PCR). A total of 2160 miRNAs were detected: 1223 were known, 937 were newly discovered and unnamed, of which 191 were expressed in all donors. A total of 7, 5, and 17 differentially expressed microRNAs (DEMs) were found in Group A vs B, Group B vs C, and Group A vs C comparisons, respectively. Twenty-two miRNAs were statistically correlated with age. Twelve miRNAs were identified as age-associated miRNAs, including hsa-miR-127-3p, mmu-miR-5100_L+2R-1, efu-miR-9226_L-2_1ss22GA, cgr-miR-1260_L+1, hsa-miR-652-3p_R+1, pal-miR-9993a-3p_L+2R-1, hsa-miR-7977_1ss6AG, hsa-miR-106b-3p_R-1, hsa-miR-186-5p, PC-3p-59611_111, hsa-miR-93-3p_R+1, and aeca-mir-8986a-p5_1ss1GA. There were 9165 target genes of age-associated miRNAs. Gene Ontology (GO) analysis of the target genes identified revealed enrichment of protein binding, membrane, cell cycle, and so on. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of age-related miRNAs for target genes revealed 139 enriched pathways, such as signaling pathways regulating stem cell pluripotency, metabolic pathways, and the Hippo signaling pathway. This suggests that miRNAs play a key role in male fertility changes with increasing age and provides new evidence for the study of the mechanism of age-related male fertility decline.

Social egg freezing: Motivations, treatment experiences and the impact of Covid-19 - a single-center experience.

This study aims to assess the motivations and treatment experiences of women undergoing social egg freezing and to understand the impact of the Covid-19 pandemic. Between January 2011 to December 2021, 191 social egg freezing patients were recruited from the Lister Fertility Clinic, London UK. Participants completed a validated questionnaire investigating patients' perspectives of social egg freezing. A response rate of 46.6% was achieved. In all, 93.9% of women expressed concern regarding age-related fertility decline which influenced their decision to undergo social egg freezing. The majority (89.5%) of women were not in a relationship at the time of social egg freezing and considered this a motivating factor. Also, 39.0% of participants had side effects related to treatment which affected work and social life. Participants were significantly more likely to experience side effects if they underwent multiple egg freezing cycles (χ2 , p < 0.01) or if they cryopreserved oocytes during the COVID-19 pandemic (χ2 , p < 0.05). Of the women, 64.0% wished to have cryopreserved oocytes at a younger age, a view significantly more likely if older than 37 years at first social egg freezing cycle (χ2 , p < 0.001). Also, 82.3% of women reported their decision to undergo social egg freezing was not delayed due to concerns regarding COVID-19 exposure during treatment; 44.1% considered the pandemic made them more willing to undergo social egg freezing. Most participants did not regret their decision to undergo social egg freezing but the majority wished they had cryopreserved oocytes at a younger age. This highlights the importance of early education to optimize outcomes and patient choice. The egg freezing process can be stressful, women may have concerns around social egg freezing and unprecedented situations such as the COVID-19 pandemic may alter treatment experience.

Assessing knowledge regarding fertility and attitude and intentions towards future parenthood among undergraduate medical students in Karachi

This study aimed to assess the knowledge regarding female fertility, infertility treatments and the attitudes regarding parenthood of medical students in Pakistan. Delayed childbirth among medical trainees due to extended years of medical education and training puts this population at a higher risk for involuntary childlessness later in life due to age-related decline in female fertility. A knowledge, attitude and practice study was carried out among medical students in Karachi in July 2021 using the English version of the Swedish Fertility Awareness questionnaire, which has been used in similar studies. Most participants wished to have children at some point in the future. However, a majority of students did not have sufficient knowledge regarding age-related decline in female fertility and overestimated the efficacy of infertility treatments. The results of this study indicate that despite planning to have children and placing great importance on parenthood, medical students severely overestimate female fertility and plan to start having children at an age at which female fecundity has begun to decline. These findings highlight a need for better provisions regarding fertility knowledge in the curriculum for medical students, as they are an at-risk group for involuntary childlessness due to age-related fertility decline.