bluuebunny/crossref_metadata_2025 huggingface dataset:
P-085 Reactive molecules drive sperm capacitation via protein posttranslational modifications
Authors: Řimnáčová H , Iniesta Cuerda M , Moravec J , Nevoral J
Published: June 2023

Study question

Are S-sulfenylation and S-nitrosylation, two essential posttranslational modifications (PTMs), increased during sperm capacitation? Which proteins are subjected to S-sulfenylation?

Summary answer

S-sulfenylation and S-nitrosylation increase during sperm capacitation. S-sulfenylation affects certain proteins, including A-kinase anchor protein 13, Fibrilline-1, and Spermatogenesis-associated protein 7.

What is known already

There is a significant increase in reactive oxygen species (ROS) and reactive nitrogen species (RNS) production within sperm capacitation. In contrast to tyrosine phosphorylation, a well-established mark of sperm capacitation, ROS, and RNS lead to PTMs themselves and modify thiol groups of cysteines. Despite these facts, the dynamics of ROS/RNS-derived PTMs during sperm capacitation remain unknown. Therefore, we consider both S-nitrosylation and S-sulfenylation of sperm proteins as novel modifications associated with the capacitation.

Study design, size, duration

C57BL6 mouse sperm were isolated from the caudae epididymae of 12-20-week-old males (n = 5). The study compared non-capacitated and capacitated spermatozoa using proteomic profiling.

Participants/materials, setting, methods

Sperm proteins were extracted into RIPA buffer. The protein lysate was incubated with 5mM dimedone, a hapten that specifically labels S-sulfenylated proteins, and immunogenically detected with an anti-dimedone antibody (Merk, Germany). Alternatively, S-nitrosylated proteins were detected by the anti-nitrosocysteine antibody (Merk, Germany). S-sulfenylated and S-nitrosylated proteins were visualized by western blotting. S-sulfenylated proteins were simultaneously identified by mass spectrometry.

Main results and the role of chance

We demonstrated an increase in S-sulfenylation and S-nitrosylation along with sperm capacitation. In addition, both S-sulfenylation and S-nitrosylation are positively correlated with tyrosine phosphorylation. Using mass spectrometry, we identified S-sulfenylated proteins, mostly belonging to the cytoskeleton: A-kinase anchor protein 13, Fibrilline-1, Spermatogenesis-associated protein 7, and Rab-like protein 2A.

Limitations, reasons for caution

The limitation of this study is the instability of PTMs. ROS/RNS-derived PTMs hypothetically arise and cease during protein sample preparation. The experiment was limited to one animal model. Further experiments should be performed on other mammalian models.

Wider implications of the findings

Studying novel sperm PTMs and their mechanism of action may improve the diagnosis of male infertility. Further, it could bring a novel approach to modulate the fertilizing ability of sperm used for in vitro fertilization.

Trial registration number

260 536
P-151 In women with recurrent implantation failures, can hyaluronan enriched media optimize reproductive outcomes?
Authors: Thorat R , Mantravadi K , Gedela D R
Published: June 2023

Study question

To evaluate the efficacy of Hyaluronan Enriched Media (HEM) to optimize Reproductive Outcomes in women with Recurrent Implantation Failures (RIF)?

Summary answer

In women with RIF, HEM seems to offer beneficial and safer reproductive outcomes.

What is known already

HEM has shown to enhance the implantation process of an embryo in-utero. Efficacy of HEM to optimize reproductive outcomes is still not proven.

Study design, size, duration

This retrospective study comprises data of 430 women with two failed IVF cycles or had RIF undergoing infertility treatment between 2016 to 2021. Study group included Young women (<37yrs) with RIF who opted elective-single blastocyst transfer (eSET) with HEM (n = 228). Control group had young women with RIF and eSET without use HEM (n = 202). Only self-gamete cycles with No PGT-A screening were included in this study. Women with known uterine pathologies were excluded from the study.

Participants/materials, setting, methods

All women had ICSI - freeze all culture strategy. In a FET, blastocyst showing 100% survival post warming was considered transfer. In study group, embryos were incubated in pre-equilibrated HEM for 10minutes before the intended embryo transfer. In control group embryo was transferred in single step media. ET procedure done as per the clinic’s SOP. Outcomes measured: Clinical Pregnancy Rates (CPR), Implantation rates (IR), Miscarriage Rates(MR), Live Birth rates(LBR) and Neonatal outcomes.

Main results and the role of chance

Following were the outcomes measured in HEM group Vs Control group:

CPR- 66.22% Vs 61.38% (p = 0.2938)

IR – 49.56% Vs 31.68% (p = 0.0002; Significant)

MR – 11.4% Vs 16.33% (p = 0.1359)

LBR – 54.82% Vs 45.05% (p = 0.0418; Significant)

There was no significant adverse events with neonatal outcomes between the groups and data looked comparable.

Use of HEM at embryo transfer for women with RIF seemed to be a beneficial intervention. It showed significant improvement with embryo implantation and live birth. Data in this study has also looked at Neonatal outcomes. Since HEM group had no reported adverse neonatal outcomes, it not only aids in improving reproductive outcomes but also is a safe intervention and can be considered for routine clinical use for indicated cases.

Limitations, reasons for caution

Retrospective data with small and unequal sample size.

Wider implications of the findings

Efficacy of various therapies is still unknown for women who had RIF. PGT-A and ERA (invasive therapies) have been tried for such cases, but its effectiveness is still up in question. Development for newer & safer therapies to improve embryo implantation and live birth for these patients is still awaited.

Trial registration number

Not applicable
P-153 Clinical outcome of different embryo transfer strategies after late rescue ICSI procedure: A cohort study based on over 140 thousand ART cycles
Authors: Zhu X , Yang R , Tian T , Li R , Liu P , Qiao J
Published: June 2023

Study question

How effective and safe is freeze-all blastocyst transfer in combination with late rescue intracytoplasmic sperm injection (r-ICSI)?

Summary answer

Freeze-all blastocyst stage embryo transfer (ET) serves as an optimal strategy to support late r-ICSI.

What is known already

Late r-ICSI has not been widely adopted due to the time-dependent in vitro deterioration of oocyte quality and endometrial growth not being synchronised with embryo development.

Study design, size, duration

This was a retrospective cohort study. All participants received treatment between 2009 and 2019. 2,270 patients in the aggregate encountered unexpected total fertilisation failure (TFF) during 149,054 cycles of in vitro fertilisation (IVF) and adopted a late r-ICSI procedure, whilst meeting the inclusion criteria for this study.

Participants/materials, setting, methods

Patients were grouped according to transfer strategies (926 women in Group 1 underwent fresh ET, 365 women in Group 2 underwent freeze-all ET, 716 women in Group 3 experienced blastulation failure, and 263 women did not have available D3 embryos). Patients received different ET strategies after r-ICSI, with the main outcome measures included live birth rate (LBR), cumulative live birth rate (cLBR), and conservative cLBR.

Main results and the role of chance

TFF occurred in 7.4 % of all IVF cycles. Group 1 tended to be older at oocyte retrieval [33.00 (30.00–37.00) vs. 31.00 (29.00–35.00), P < 0.001], with more infertile years [5.00 (3.00–8.00) vs. 3.00 (2.00–6.00), P < 0.001], higher follicle-stimulating hormone (FSH) levels [7.10 (5.71–8.69) vs. 6.61 (5.25–7.95) mIU/mL, P < 0.001], higher gonadotropin consumption [2850.00 (1950.00–4125.00) vs. 2325.00 (1800.00–3150.00) IU, P < 0.001], and fewer oocytes retrieved [10.00 (6.00–15.00) vs. 13.00 (9.00–19.00), P < 0.001]. Group 2 exhibited considerably better LBRs following the first ET cycle (37.53 % vs. 4.64 %) and cLBR (52.60 % vs. 8.21 %). After adjustment for covariates using binary logistic regression analyses, Group 2 still showed better obstetric performance in LBR (OR:11.77, 95 % CI (8.42–16.45)], cLBR (OR:11.29, 95 % CI (7.84–16.27)], and conservative cLBR (OR:2.55, 95 % CI (1.83–3.55)]. Additionally, the two groups showed similar miscarriage rates, whilst no new-borns with malformations or congenital diseases were reported.

Limitations, reasons for caution

This study was based on a retrospective cohort, which is inevitably associated with a skewed data distribution and inherent biases.

Wider implications of the findings

Our study highlights r-ICSI as a safe and effective alternate solution for unexpected TFF to aid frustrated couples. However, for women with limited oocytes available for r-ICSI use, weighing the benefits against the costs of the procedure might be prudent before implementing in vitro blastulation.

Trial registration number

NA
P-168 Follow-up survey of deliveries derived from Day 7 blastocysts
Authors: Murakami K , Tsuji H , Kitasaka H , Fukunaga N , Asada Y
Published: June 2023

Study question

Does Day 7 blastocyst-transfer affect prognosis of babies born?

Summary answer

There was no effect on growth up to 3 years of age in babies from Day 7 blastocyst-derived births.

What is known already

Culture medium and technology have advanced, and the efficiency to culture up to the blastocyst stage in order to obtain viable embryos has increased dramatically. As a result, blastocyst transfer is associated with higher clinical pregnancy outcomes than cleavage transfer. In 1998, it was first reported that some embryos may develop into blastocysts on Day 7. However, there are few reports on neonatal outcomes from Day 7 blastocyst transfer. Therefore, the utility of Day 7 blastocysts is controversial.

Study design, size, duration

This was retrospective study which included 23 cases where there was a live birth after single embryo transfer of a Day 7-derived blastocyst. The incidence of birth defects, birth weight was recorded as well as a physical development survey. The height and weight were recorded in 10 children aged 18-months and 4 children aged 3 who responded to follow-up survey. The time period was from January 2013 to December 2021.

Participants/materials, setting, methods

Patient seeking fertility treatment at an established private IVF clinic. We compared the birth weight, and birth after 18-months and 3 years height and weight of children born to a Day 7 blastocyst with data from babies born from Day 5 and Day 6 derived blastocysts. Statistical significance was determined using Kruskal-Wallis test.

Main results and the role of chance

The incidence of birth defects was 2.4% (115/4828), 1.7% (16/954) and 0% (0/23) in the Day 5, Day 6 and Day 7 groups, respectively. The average birth weight of the Day 7 group was 3057+/-532 g, which was not significantly different from 3024+/-472 g and 3041+/-480 g in the Day 5 and Day 6 groups. The average height at 18-months, was 80.0+/-5.3 cm, 79.8+/-5.1 cm, 77.8+/-6.9 cm in the Day 5, Day 6 and Day 7 groups, respectively. The average weight at 18-months, was 10.5+/-1.2 kg, 10.4+/-1.1 kg, 10.5+/-1.4 kg. The average height at 3 years old, was 93.2+/-4.8 cm, 92.9+/-4.7 cm, 93.4+/-5.9 cm. The average weight at 3 years old, was 13.9+/-1.6 kg, 13.8+/-1.4 kg, 14.1+/-2.6 kg, respectively. There was no significant difference in average height and weight up-to the 3 years old follow-up survey.

Limitations, reasons for caution

The incidence of Day 7 blastocyst-transfer is low and the study was limited to cases of single blastocyst embryo transfer. The study was not separated by sex, because sample size was small.

Wider implications of the findings

There was no difference in growth up to a 3-year-old follow-up survey of the Day 7 blastocyst compared with the Day 5 and Day 6 groups. Transfer of a Day 7-derived blastocyst does not appear to affect infant development and should be considered in cases where blastocyst development is delayed.

Trial registration number

not applicable
P-180 Hyaluronidase used in oocyte denudation inhibits the proliferation and the viability of cumulus cells through the inhibition of midkine
Authors: Ergüven M , İrez T
Published: June 2023

Study question

Does hyaluronidase (HAse) have possible negative or positive effects on cumulus cells (CCs) during oocyte denudation before ICSI by altering midkine (MK) levels?

Summary answer

Hyaluronidase inhibits the proliferation of CCs via the inhibition of MK activity which may result in poor embryo quality and low ICSI success.

What is known already

MK is abundantly expressed in ovarian follicles. MK secreted from the cumulus-granulosa cells that surround oocytes was shown to promote the cytoplasmic maturation of oocytes. This effect of MK could be mediated via its' anti-apoptotic effect and some other mechanisms. The removal of CCs during oocyte denudation (OD) is done in order to select and grade the oocytes before ICSI. HAse is applied for ICSI to break down hyaluronic acid, which is present at high levels in the cumulus-oocyte complex during OD. To date, there have only been a few reported cases of the toxic effects of hyaluronidase on oocytes.

Study design, size, duration

This study is a prospective, randomized study done with 90 healthy women between September 2017 and September 2018.

Participants/materials, setting, methods

This study was done with women diagnosed as a male factor between the ages of 21 and 40 who underwent ICSI. HAse was applied to the cultured CCs at concentrations of 0.1 IU/ml, 1 IU/ml, and 10 IU/ml. The proliferation and apoptosis indices (Flow cytometry), structures [Transmissive electron microscopy (TEM)], and MK levels (ELISA) of CCs were evaluated every 24 hours for 48 hours. One way-Anova was used and p < 0.05 was considered statistically significant.

Main results and the role of chance

The application of HAse at all concentrations decreased cumulus cell numbers for 48 hours (p < 0.05). The highest decrease in cell number and cell viability with the highest number of apoptotic cells were detected at the 48th hour at the application of the highest concentration of HAse (p < 0.05). The highest concentration of HAse application caused the highest decrease in MK levels at the end of 48 hours (p < 0.05).

Limitations, reasons for caution

The effect of HAse on the CCs of women aged below 21 and above 40 could not be evaluated. Therefore, the overall resistance and fragility of CCs to these HAse concentrations could not be evaluated. In addition, the HAse effects should be evaluated in female infertility cases such as PCOS.

Wider implications of the findings

This is the first report to examine the effect of HAse on MK activity. 2-3 lines of CCs are commonly left during oocyte denudation in order to protect the oocyte from stress-induced reactions. The low number of CCs means low MK levels resulting in a loss in oocyte competence.

Trial registration number

Tthe Ethics Committee Directive on Non-Interventional Studies of Biruni University with the permission numbers 2017/5-1.
P-181 The impact of high proportion of immature oocyte in a cohort on the reproductive outcome following icsi
Authors: Kant G , Nayar K D , Prasad A , Sanan S , Miglani S , Sharma H , Nayar K
Published: June 2023

Study question

Whether high proportion of immature oocyte in a cohort impacts outcome of sibling mature oocyte in ICSI cycles.

Summary answer

Study demonstrate that if immature oocyte retrieved was more than 50% in a cycle then it reduces reproductive outcome with the remaining mature sibling oocytes.

What is known already

The selection of competent oocyte is crucial to assisted reproductive technique procedure outcome. In ICSI procedure where only metaphase II oocytes (MII) are injected correlates with number of MII oocytes available, embryo development and pregnancy outcome. Percentage of immature oocyte {Metaphase I (MI) and Germinal Vessicle (GV) } retrieved in a cycle is always a concern for stimulation protocol, patient response which affects IVF laboratory outcome.

Study design, size, duration

A prospective study was conducted from 1st January 2020 to 1st December 2022 in the Department of Reproductive medicine at a tertiary infertility centre in India.

Participants/materials, setting, methods

Patients were divided into 3 groups on the basis of percentage of immature oocyte retrieved in a single cycle. Group A (n = 100), where percentage of immature oocyte (MI and GV) was between 0 to 20%, while Group B (n = 60) where percentage of immature oocyte was between 21 to 50% and Group C (n = 20) where percentage of immature oocyte was between 51 to 100%.

Main results and the role of chance

This study included all normal responder patients and cycles using in-vitro fertilisation (IVF) and poor responders were excluded. All blastocyst formed were vitrified and transferred in frozen embryo replacement cycle. All the 3 groups were compared on the basis of fertilisation rate, blastocyst formation rate, clinical pregnancy rate and miscarriage rate.

Group A and group B had no significant difference in fertilisation rate (82% vs. 78%, p > 0.5), blastocyst formation rate (60% vs. 58%, p > 0.5), clinical pregnancy rate (58% vs. 55%, p > 0.5) and miscarriage rate (12% vs. 11%, p > 0.5).

While significant reduction was observed in group C ( > 50% MI & GV oocyte) in fertilisation rate (40%, p < 0.1), blastocyst formation rate (28%, p < 0.1), clinical pregnancy rate (15%, p < 0.1).

Limitations, reasons for caution

Larger randomised control studies are needed to strengthen these results.

Wider implications of the findings

Percentage of immature oocyte can help in predicting the success of assisted reproductive technique. This will help in counselling patient about outcome.

Trial registration number

NA
P-194 Initial experience of using iDAScore as a tool to predict euploid blastocysts
Authors: Tan S H , Lim A Y X , Khoo C L , Yap W Y , Keith J M , Lee C S S
Published: June 2023

Study question

Can the annotationfree embryo scoring system (iDAScore) predict the likelihood of euploidy in embryos?

Summary answer

Our results have shown that there is a significant positive correlation between iDAScore and blastocyst euploidy.

What is known already

iDAScore (Vitrolife, Sweden) is an embryo ranking model that was developed by using artificial intelligence (AI) and deep learning. It does not require any user-dependent annotation which largely eliminates the subjectivity of manual annotation by embryologists. It has also been reported to be a robust predictor in likelihood of embryo implantation. However, the correlation between iDAScore and euploidy has not been extensively studied. Therefore, this study is to determine if iDAScore can be utilized as a predictor for embryo euploidy.

Study design, size, duration

A total of 860 blastocysts (mean age: 30.0; age range: 19.0-35.0) were assessed with iDAScore on day-5 and/or day-6 of embryo development from January 2022 to October 2022 at Alpha IVF & Women’s Specialists. The euploidy rates were analyzed based on the stratified iDAScores (Group A:0.0-4.9; Group B: 5.0-7.4; Group C:7.5-8.9; Group D:9.0-10.0).

Participants/materials, setting, methods

Scores were computed by the iDAScore software at the Vitrolife Technology Hub (Vitrolife, Gothenburg, Sweden) prior to trophectoderm biopsy for Preimplantation Genetic Testing for Aneuploidies (PGT-A) (Ion Torrent, USA). The ploidy status (euploid or aneuploid) based on PGT-A results were recorded and assessed with the corresponding iDAScore.

Main results and the role of chance

The euploidy rates of Group A, B, C and D were 56.7% (17/30), 60.0% (105/175), 65.9% (178/270) and 74.8% (288/385) respectively. Group D shows significantly higher euploidy rate compared to Group B and Group C (p = 0.0005, p = 0.0143). A weak significance was observed between Group D and Group A (p = 0.0505). This is probably due to smaller sample size in Group A. No significant differences were displayed among Group A, Group B and Group C (p = > 0.005). Our results demonstrated an increasing trend between iDAScore and the proportion of euploid embryos.

Limitations, reasons for caution

Few of the blastocysts in Group A could be selected for biopsy so the sample size was only a fraction of the other groups. Further studies and larger sample sizes would allow the findings to be stated more confidently.

Wider implications of the findings

Although the iDAScore algorithm scores the chance of embryo implantation, we have demonstrated its potential to assist in predicting the embryo ploidy status in a non-invasive manner. This may be used to inform the decision of which and how many blastocysts to biopsy and transfer, thus reducing treatment costs.

Trial registration number

not applicable
P-196 PIEZO-intracytoplasmic sperm injection (P-ICSI) can improve the clinical outcomes of ICSI
Authors: Lim C K , Cha H J , Shin S H , Kim G W , Ryu H , Han S , Choi W
Published: June 2023

Study question

Can P-ICSI improve the clinical outcomes of ICSI, such as degeneration, fertilization and development to blastocyst, compared to conventional ICSI (C-ICSI)?

Summary answer

P-ICSI can improve the clinical outcomes of ICSI by increasing the number of fertilized oocytes and decreasing the number of degenerated oocytes after ICSI.

What is known already

In human IVF-ET, some of oocytes are damaged after ICSI when retrieved oocytes are inseminated by ICSI. The application of P-ICSI has been limited although it was developed over 30 years ago. Recently, it has been reported that P-ICSI can decrease damage after ICSI and improve fertilization and embryonic development compared to C-ICSI.

Study design, size, duration

This study included 50 subjects undergoing IVF and was performed between April and October 2022. Sibling oocytes were randomly assigned to C-ICSI and P-ICSI.

Participants/materials, setting, methods

A total of 1,207 oocytes were retrieved from 50 patients. Among them, 852 oocytes with visible spindle were inseminated by C-ICSI or P-ICSI in sibling oocytes. Four hundred twenty-five oocytes were inseminated by C-ICSI and 427 oocytes were by P-ICSI. Fertilization, degeneration after ICSI and blastocyst development on day 5 or day 6 were compared between the P-ICSI and C-ICSI. The differences between C-ICSI and P-ICSI were statistically analyzed using Mann-Whitney U test.

Main results and the role of chance

Normal fertilization rate of P-ICSI (78.2 ± 16.5%) were higher than that of C-ICSI (71.2 ± 18.9%). The difference was statistically significant (P = 0.0447). The oocyte degeneration rates after ICSI were 9.3 ± 11.8% (C-ICSI) and 4.8 ± 9.4% (P-ICSI). The degeneration rate of C-ICSI was significantly higher than that of P-ICSI (P = 0.04338). Blastocyst formation rate of C-ICSI was 58.6 ± 25.0% and that of P-ICSI was 64.3 ± 26.3%. The blastocyst formation rate of P-ICSI was higher than that of C-ICSI but the difference was not statistically significant (P = 0.2113). The rates of blastocysts that were available for embryo replacement or cryopreservation were higher in P-ICSI (56.0 ± 25.4%) than in C-ICSI (50.4 ± 24.7%) although the difference was not significant (P = 0.26272).

Limitations, reasons for caution

Pregnancy outcomes data were not available. The data of this study were obtained from single fertility center. More large-scale multicenter studies will be needed to confirm the effectiveness of P-ICSI.

Wider implications of the findings

Consistent with other studies, these results show that P-ICSI reduces the damage rate of oocytes after ICSI. The implementation of P-ICSI has the potential to improve clinical outcomes of human IVF although it is more complicated and takes longer than that of C-ICSI.

Trial registration number

not applicable
P-211 “Survival rates after vitrification of oocytes obtained after luteal phase vs follicular phase ovarian stimulation”
Authors: Lopez Feijoo C , Cerrillo M , Kohls G , Bronet F , Molina M J , García Velasco J A
Published: June 2023

Study question

Do the oocytes retrieved after luteal phase ovarian stimulation (OS) have the same survival rate as those retrieved after follicular phase (OS)?

Summary answer

Oocyte survival rate after vitrification is similar in oocytes retrieved in a follicular phase OS compared to those in luteal phase.

What is known already

Oocytes retrieved in luteal phase have recently captured scientific interest as dual stimulation is gaining popularity. Oocytes obtained in luteal phase OS added to those obtained in follicular phase shorten time to pregnancy per intention to treat in poor responders. Embryos obtained with these oocytes show similar ploidy rates, as well as clinical outcomes.

Study design, size, duration

We analyzed data from a randomized, non-inferiority Duo Stim Preimplantation Genetic Testing for aneuploidies (PGT-A) trial performed from 2017 to 2021 in women >38 years old. Only cycles with at least one oocyte retrieved in both follicular and luteal were included.

We compared survival rates from oocytes obtained in after follicular phase OS (n = 154 oocytes) versus luteal phase OS (n = 201 oocytes) within the same patient.

Participants/materials, setting, methods

A subtotal of 38 patients were included who underwent 2, 3, or 4 OS cycles to accumulate oocytes for subsequent ICSI with PGT-A. After retrieval and decumulation, oocyte vitrification was performed with Kitazato vitrification protocol (Kitazato, Japan) and Cryotop device, same as subsequent thawing.

Main results and the role of chance

Mean of age in the selected patients was 39,7 years. In total, 195 cumulus-oocyte complexes were retrieved after follicular phase OS vs 245 in luteal phase. From this group of oocytes, 145/195 were mature (78.97%) vs 201/245 (82.04%), respectively.

No statistically significant differences were observed in survival rate whether oocytes came from a follicular phase OS 81.16 % (125/154) versus luteal phase 87.06% (175/201) (p = 0.3312).

Our results confirm that survival rate of oocyte obtained in a luteal phase OS is comparable to conventional follicular phase OS, validating oocyte accumulation strategy in a Duo Stim protocol.

Limitations, reasons for caution

The limited sample size was due to the nature of the randomized, controlled trial. Our results need to be replicated in larger series for validation.

Wider implications of the findings

Luteal phase OS oocytes showed similar survival rates after thawing than conventional OS cycles, opening a new venue for oocyte vitrification in oncological patients or poor responders accumulating oocytes. This approach might be of interest as well in patients or countries where ethical/legal limitations may limit embryo accumulation.

Trial registration number

not applicable
P-231 The effect of warming blastocysts in Vitrolife media previously vitrified in Cook media on reproductive potential and clinical outcome
Authors: Withey A , Craig J , Watkins J
Published: June 2023

Study question

Does the use of a different manufacturer’s media for vitrification and warming of blastocysts affect pregnancy, miscarriage and live birth rates.

Summary answer

Warming blastocysts in Vitrolife media, previously vitrified in Cook media, does not significantly affect the pregnancy or live birth rate.

What is known already

Current good practice is to vitrify and warm blastocysts in media manufactured by the same company. This is due to the optimisation of the protocols and media composition intending to maintain high survival rates and associated pregnancy rates. However, circumstances can arise, such as supply and production issues, whereby it is not possible to source the same media for warming vitrified blastocysts as the media they were vitrified in.

Study design, size, duration

This retrospective cohort study was conducted at TFP-Oxford Fertility. Frozen embryo transfer (FET) cycles included in the study were performed between 2018 and 2022. Embryos that had undergone genetic testing were excluded from the study. The mean women's age was 35 in the control group (Cook warming media) and 36 in the study group (Vitrolife warming media). The average number of embryos transferred was 1.06 vs 1.17 in control vs study group.

Participants/materials, setting, methods

For the present study 817 patients underwent FET between 2016 and 2018 whereby blastocysts were vitrified in cook media (Sydney IVF Blastocyst Vitrification Kit, K-SIBV-5000) and warmed in cook media (Sydney IVF Blastocyst Warming Kit, K-SIBW-5000). In comparison 704 patients underwent a FET between 2018 and 2022 whereby blastocysts were vitrified in cook media and warmed in vitrolife warming media (Rapidwarm™ Blast, 10120). The pregnancy, clinical pregnancy and live birth rates were compared.

Main results and the role of chance

Fisher’s exact test was used to calculate the p value and a level of 0.05 was used for significance. The results showed that there was no significant difference in the pregnancy rate, clinical pregnancy rate or live birth rate for blastocysts warmed in Vitrolife media vs Cook media (59.0% vs 57.0% p = 0.56, 47.2% vs 43.9% p = 0.44, 39.3% vs 37.0% p = 0.72)

Limitations, reasons for caution

This is a retrospective study at a single centre. The number of previous treatment cycles per couple was not included in the study. The study only looked at a comparison of Cook and Vitrolife warming media, no other commercially available media was tested.

Wider implications of the findings

Although good practice to vitrify and warm blastocysts in the same manufacturer media, our results show that the reproductive potential and clinical outcome may not be adversely affected by using a different manufacturer’s media. However, composition of the media and protocol should be considered and further studies should be conducted.

Trial registration number

not applicable
P-255 Increased time in thawing solution improves human oocyte warming outcomes
Authors: Schlenker T , Schoolcraft W , Swain J
Published: June 2023

Study question

Can increased time in thaw solution improve oocyte survival and outcomes following vitrification/warming?

Summary answer

Exposing vitrified oocytes to the first step thawing solution for 90 seconds improved oocyte survival compared to 60 second exposure.

What is known already

Oocytes are a sensitive cell type and careful handling of the cells is required to obtain high outcomes following vitrification and warming. Carefully timed exposure to various vitrification solutions is required to avoid toxicity and ensure proper vitification. Similarly, proper and carefully timed exposure to warming solutions is required to avoid oocyte damage upon warming. Recent reports suggest a modified and rapid warming protocol may be beneficial for vitrified blastocysts. However, oocytes are unique cells and may require unique considerations to optimize warming.

Study design, size, duration

Retrospective analysis of donor oocyte warming outcome data within a single IVF lab over a 12 month period comparing a 60 second first step during oocyte warming (control n = 637 oocytes) versus a very consistent 90 second exposure (n = 672 oocytes) to the first step thawing solution.

Participants/materials, setting, methods

Human donor oocytes were vitrified using the standard Kitazato protocol with ES drop merging and loading on CryoTop devices using the wicking method prior to plunging into liquid nitrogen. Warming was performed using the standard protocol of 60 seconds in thaw solution (TS) or 90 seconds exposure. All cycles utilized ICSI and excluded testicular sperm samples. Survival, fertilization and blastocyst development were compared between the two exposure groups. Data were analyzed utilizing Fisher’s Exact Test.

Main results and the role of chance

Oocyte survival differed between the two timing treatments, with exposure to TS for 90 seconds yielding higher oocyte survival (96.1%) than 60 secondsTS exposure (92.0%), p < 0.002. No differences in fertilization following ICSI (78.3.0% vs. 79.0%) orin good quality blastocyst conversion (≥3BB) on day 5 (22.1% vs. 25.1%) or by day 7 (55.3% vs. 55.7%) were apparent. The aneuploidy rate of blastocysts were similar, with 28.5% in the 90 second TS treatment group and 27.3% in the 60 second TS exposure group.

Limitations, reasons for caution

Data were examined retrospectively and sibling oocyte splits were not utilized. Clinical outcomes following resulting embryo transfer were not examined.

Wider implications of the findings

These data may help improve oocyte warming protocols and resulting outcomes.

Trial registration number

not applicable
P-307 Is multinucleation as bad as we ever thought?
Authors: Granell Martínez M
Published: June 2023

Study question

Could the presence of multicleation at early stages influence further embryo development?

Summary answer

The presence of multinucleation at 2 cell stages is independent on the success, but not at 4 cell stage.

What is known already

The presence of multinucleated blastomeres (MNB) in the cleaving embryo has been linked to poor embryo development and adverse in vitro fertilization (IVF) outcomes, when detecte on days 2 and 3 of embryo culture under inverted microscope inspections once a day. Multinucleation has been associated with a lower rate of blastocyst formation, increased rates of aneuploidy and chromosomal abnormalities, leading to chromosomal chaos and a decreased rate of implantation and arrested embryo development. With the introduction of Time-Lapse (TL) technology, it is been demostrated that multinucleation at the 2 cell stage is a very frequent issue.

Study design, size, duration

It is a retrospective observational study. 195 embryos from 28 patients that underwent and IVF treatment at the Unit of Medicine Reproductive of the Hospital General Universitario de Valencia were included in this study from 2017 to 2021 to study embryo development. Those patients whose embryos could be visualized with quality in a Time-lapse incubator were selected.

Participants/materials, setting, methods

The study of the 195 embryos from the 28 patients was carried out in the MIRI® TL6 Time-Lapse incubator. These embryos were checked at the 2-cell, 4-cell and blastocyst stages. The Shapiro-Wilk test was used to determine whether the sample had a normal distribution. and the Pearson test was used to determine if there were significant differences between the samples. Besides, the Chi-square test was used to determine the correlation between 2 qualitative variables.

Main results and the role of chance

108 fertilized embryos were studied, 78.7% of them did develop blastocyst formation; 50% did not show multinucleation and 28.7% did, the analysis indicated that there was no correlation between blastocyst formation and the presence of multinucleation at 2 cell-stage (p = 0.364). The analysis indicated that multinucleated cells showed a prolongation of the division time, the analysis indicated that it had a normal distribution (p < 0.001) and it exist significant differences (p < 0.001). Pregnancy was obtained in 5% and 9% of the embryos with multinucleation at 2 cell-stages. Finally, analysis of autocorrections on day 2 showed that these occurred in 60% and 40% of 2 cell-stage.

Limitations, reasons for caution

As these data correspond to patient profiles from our unicenter study, the data could be different in other assisted reproduction centers. In addition, a larger study sample size is necessary in order to confirm our results.

Wider implications of the findings

These results can help us to decide the fate of the embryos and not to penalize those embryos that present multinucleations at the 2-cell stage.

Trial registration number

not applicable
P-322 Single-cell transcriptome analysis reveals endometrial immune microenvironment in minimal/mild endometriosis
Authors: Huang X , Huang W
Published: June 2023

Study question

This study aimed to systematically understand the endometrial leukocyte types, inflammatory environment, and impaired receptivity at single-cell resolution.

Summary answer

These results provide new insights into the endometrial immune microenvironment and impaired endometrial receptivity in infertile women with minimal/mild endometriosis.

What is known already

Endometriosis is a common inflammatory disorder in women of reproductive age due to an abnormal endometrial immune environment and is associated with infertility.

Study design, size, duration

We profiled single-cell RNA transcriptomes of 138,057 endometrial cells from endometriosis patients (n = 6) and control (n = 7), respectively, using 10x Genomics platform.

Participants/materials, setting, methods

We profiled single-cell RNA transcriptomes of 138,057 endometrial cells from endometriosis patients (n = 6) and control (n = 7), respectively, using 10x Genomics platform.

Main results and the role of chance

We found that one cluster of epithelial cells that expressed PAEP and CXCL14 was mostly from the control during the window of implantation (WOI). This epithelial cell type is absent in the eutopic endometrium during the secretory phase. The proportion of endometrial immune cells decreased in the secretory phase in the control group, whereas the cycle variation of total immune cells, NK cells, and T cells was absent in endometriosis. Endometrial immune cells secreted more IL-10 in the secretory phase than in the proliferative phase in the control group; the opposite trend was observed in endometriosis. Proinflammatory cytokines levels in the endometrial immune cells were higher in endometriosis than in the control group. Trajectory analysis revealed that the secretory phase epithelial cells decreased in endometriosis. Ligand–receptor analysis revealed that 11 ligand-receptor pairs were upregulated between endometrial immune and epithelial cells during WOI.

Limitations, reasons for caution

Our study had some limitations, such as the small sample size and lack of validation experiments. Further studies are needed to demonstrated the conclusions of our bioinformatic analysis, like in situ validation of receptor-ligand pairs analysis. Therefore, more evidence is needed to clearly state the pathogenesis of endometriosis-associated infertility.

Wider implications of the findings

This database will provide an essential resource for understanding the eutopic endometrial inflammatory environment and defective endometrial receptivity in patients with endometriosis-associated infertility.

Trial registration number

82071625
P-414 Trends in patient age at planned oocyte cryopreservation
Authors: Hasson J , Shavit T , Hyman J , Tsafrir A
Published: June 2023

Study question

Did the age of women who underwent Planned Oocyte Cryopreservation (POC) change in the last decade?

Summary answer

Mean age of women presenting for POC decreased from 38.2±2.8 to 35.6±2.7.

Proportion of women <36 increased from 13% in 2011 to 51% in 2022.

What is known already

Mean age of women who had POC reported in the literature to date is 37-8.

Given that efficacy of POC is strongly related to age, some clinicians advice women considering POC to complete the procedure before age 35.

Study design, size, duration

Retrospective, observational multicenter study of all POC cycles from January 2011 to October 2022. Women who had oocyte cryopreservation for other indications were excluded.

Participants/materials, setting, methods

In all, 4021 women had at least one POC cycle in two large private IVF units belonging to the same medical organization in Israel. The main outcome measure was age at first cycle. Total number of women who underwent POC at each year and age at first cycle of every woman were recorded. Data was analyzed using SPSS version 23. ANOVA was used to test whether mean age differed significantly throughout the years.

Main results and the role of chance

Since 2011, when POC was first approved in Israel, until the end of October 2022, 4021 women underwent at least one cycle of POC in our units. The number of women who had POC per year increased gradually from 62 in 2011 to 979 in 2021 (last year with full data).

Mean age declined with each year, from 38.2±2.8 in 2011 to 35.6±2.7 in 2022. The proportion of women aged < 36 increased gradually from 13% in 2011 to 35% in 2019 and further to 51% in 2022 (R = 0.20, p value<0.01).

Limitations, reasons for caution

This study was performed in a high-socioeconomic catchment area of one country. This may limit the generalizability of our findings to other populations. In addition, the usage rate and delivery rates of these patients are not known yet.

Wider implications of the findings

The significant decrease in age of women presenting for POC and the increased proportion of women<36 may present a growing awareness for age related infertility and possibly increased efficacy of POC in younger women.

Trial registration number

not applicable
P-451 Fertility awareness among Tunisian university students: A state-of-knowledge survey
Authors: Daoud S , Maalej M , Abdelkefi O , Benamor M , Sellami A , Chaabene K , Rebai T
Published: June 2023

Study question

To investigate the beliefs and knowledge of university students regarding fertility.

Summary answer

University students are aware of the impact of unhealthy lifestyle on fertility. There are however misconceptions about the ideal age for childbearing and treatment limits.

What is known already

The gradual decline in fertility is currently well documented. Furthermore, the mean age of first-time mothers is increasing, especially in groups with higher educational levels. However, the level of awareness (sensitization) of couples to infertility factors such as female age and environmental factors is not well-known.

Study design, size, duration

A 12 items-questionnaire on infertility aspects « 12QSI » was previously developed and validated. The questionnaire is divided into 3 sections: (i) epidemiology of infertility, (ii) causes and risk factors for infertility, and (iii) treatment of infertility. A seven-month online survey of a random sample of Tunisian students was conducted. Data collection was carried out via Google Forms® by sharing the link on social media platforms and groups.

Participants/materials, setting, methods

The survey was completed by 800 respondents. We retained 775 responses according to the inclusion criteria: complete response, being of childbearing age and attending university education. Statistics were obtained using SPSS® software.

Main results and the role of chance

Participants were 24.6 ± 5 years of age on average. Almost 57% were women. The average knowledge score was 56.7 ± 28%. The lowest scores were found for the questions on infertility treatment (only 15% of correct answers) with an overestimation of the chances of success of Assisted reproductive technology (ART). An underestimation of the impact of age on both male and female fertility was reported. The mean score was significantly better for the group of respondents studying medicine than for the rest (63 ± 32% vs. 48 ± 26%; p < 0.001), and for respondents who have faced an infertility problem compared to those who have not (65 ± 28% vs. 56.3 ± 29%; p < 0.001).

Limitations, reasons for caution

The representativeness of the sample is not ensured. Survey distribution only through social networks could be a limit.

Wider implications of the findings

Over-optimism regarding the importance of age and the effectiveness of infertility treatments is noted, which can lead to unintended infertility. Educational programs should be developed to address erroneous beliefs and to increase the population`s knowledge of fertility.

Trial registration number
P-470 Prediction of the cumulative live birth rate of IVF using a human embryonic stem cells-derived trophoblastic spheroid implantation model
Authors: Ng E , Cherie L , Ruan H , Lee K C , Fong S W , Yue C , Chen A , Lee C , Lam M T , Yeung W , Li R
Published: June 2023

Study question

Can the attachment rate of a human embryonic stem cell-derived trophoblastic spheroids (BAP-EB) onto endometrial epithelial cells predict the cumulative live birth rate of an IVF cycle?

Summary answer

The BAP-EB attachment rate offers modest prediction of the cumulative live birth rate of women aged ≥35 years undergoing IVF.

What is known already

Implantation failure is one of the major causes leading to a low IVF success. There is no reliable tool for assessing endometrial receptivity in IVF.

Study design, size, duration

Prospective observational study of 240 infertile women from 2017 to 2021.

Participants/materials, setting, methods

Infertile women with regular cycles attending IVF centres were recruited. Endometrial aspirates were collected from a natural cycle one month before IVF for the BAP-EB attachment rate. The cumulative live birth rate of a stimulated cycle and its derived frozen embryo transfer cycles within 6 months of ovarian stimulation were obtained.

Main results and the role of chance

The BAP-EB attachment rate in women with a cumulative live birth was similar to those who failed to have a live birth. When women were stratified by age into <35 years and ³35 years, the BAP-EB attachment rate was significantly higher only in women aged ³35 years who had a live birth when compared to those without a live birth. The receiver operating characteristic (ROC) curve analysis of BAP-EB attachment rate in predicting cumulative live birth showed the area under the curve (AUC) of 0.559 (95% CI 0.479 – 0.639, p = 0.151), 0.448 (95% CI 0.310 – 0.585, p = 0.461) and 0.613 (95% CI 0.517 – 0.710, p = 0.026) for all ages, age <35 and ≥35 respectively.

Limitations, reasons for caution

BAP-EB and the isolated endometrial epithelial cells may not fully represent the in vivo developed human blastocysts and endometrial cells, respectively. The in vitro nature of the experiments and the limited sample size in this study may also limit the interpretation of the data.

Wider implications of the findings

Implantation failure is one of the major reasons contributing to the low success rate of IVF. The BAP-EB-endometrium attachment assay could potentially be used by reproductive medicine specialists to better counsel the couples who fail in IVF treatment.

Trial registration number

NCT02713854