2PN Oocytes Research Articles

Evaluation of the follicular fluid thiol/disulfide balance among patients with poor ovarian response.

This study aimed to compare the thiol/disulfide balance, myeloperoxidase, and ischemia-modified albumin levels in the follicular fluid (FF) of poor ovarian response (POR) and normal ovarian response (NOR) women who received intracytoplasmic sperm injection (ICSI). The study was performed between March 2021 and April 2022 at the Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Ankara City Hospital. The study included 27 POR and 35 NOR women who underwent ICSI. FF was obtained after the controlled ovarian stimulation cycle. The FF thiol/disulfide balance was detected using spectrophotometric methods. A correlation analysis was conducted to determine whether these oxidative stress markers could contribute to predicting oocyte quality. Disulfide levels were significantly higher in the NOR group than in the POR group (p=0.014). The number of fertilized egg (2PN) oocytes was positively correlated with the total thiol level (r=0.258, p=0.046). The disulfide level was positively correlated with the anti-Müllerian hormone level (r=0.262, p=0.039) and the total number of retrieved oocytes (r=0.335, p=0.008). The disulfide levels differed significantly between the NOR and POR groups. The statistically significant differences of fewer metaphase II oocytes and lower percentage of good-quality embryos in the NOR group compared to the POR group might have resulted from the NOR group's elevated disulfide levels. The total thiol levels correlated with the total of 2PN oocytes. Future studies should examine the thiol/disulfide balance at assisted reproductive technology centers to predict which oocytes could be fertilized.

P-383 Development and validation of a visualized prediction model for early miscarriage risk in patients undergoing IVF/ICSI procedures: a real-world multi-center study

Abstract Study question What are the key predictors of early miscarriage in patients undergoing IVF/ICSI treatment? Summary answer This study identified risk factors for early miscarriage in IVF/ICSI patients, and developed a validated predictive nomogram, highlighting age-dependent factor variations. What is known already Early miscarriage, occurring in 15-20% of confirmed pregnancies, can lead to complications, psychological distress, and fertility impairment. It is especially prevalent during the first trimester, with up to 80% of all miscarriages happening during this phase. Women who conceive via Assisted Reproductive Technology (ART) face higher stress levels and increased risk of miscarriage. Although many contributing factors to early miscarriage have been identified, there is a scarcity of integrated predictive models for IVF/ICSI patients. This paper aims to develop a validated, user-friendly nomogram to predict first-trimester miscarriage risk in IVF/ICSI patients and help clinicians and patients make informed decisions. Study design, size, duration A retrospective analysis was conducted on 20,322 first cycles out of 31,307 for IVF/ICSI at a reproductive centre in Southern China, After excluding ineligible cycles, 6,724 first fresh cycles were included and randomly divided into a training dataset (n = 4,516) and an internal validation dataset (n = 2,208). An external validation dataset (n = 1,179) was obtained from a reproductive center in Northern China, further ensuring the robustness and generalizability of the findings. Participants/materials, setting, methods This retrospective study analysed 6,724 out of 31,307 IVF/ICSI treatments from 2011 to 2020 at a Southern China reproductive center and 1,179 cycles from a Northern China center (2013-2018) for validation. Early miscarriage was defined as a loss before 12 weeks’ gestation. Patients were divided into two age groups: ≥35 and <35 years. The study followed the TRIPOD statement and received Institutional Review Board approval. Main results and the role of chance The study found statistically significant differences across the training, internal validation, and external validation sets in terms of patient age, BMI, number of good quality embryos, antral follicle count (AFC), basal testosterone (T), luteinizing hormone (LH) levels, among other variables. The early miscarriage rates were 9.43%, 10.55%, and 13.50% across these sets. Elder women (age ≥35 years) had a higher duration of infertility and early miscarriage rate, but lower AFC and numbers of retrieved oocytes, 2PN, and MII oocytes compared to younger women (age <35 years). A nomogram prediction model was developed using variables such as female age, BMI, number of spontaneous abortions, number of induced abortions, and basal FSH, which were found to be independent prognostic factors for early miscarriage. The model’s performance was validated using AUC, calibration plots, and DCA, indicating acceptable performance and good agreement between prediction and observation. The model was more beneficial for predicting miscarriage risk than the treat-all or treat-none schemes. Subgroup analysis for elder and younger women showed different risk factors for miscarriage, notably female age, basal FSH for elder women, and female age, BMI, and the number of spontaneous abortions for younger women. The subgroup models also showed consistent results and good clinical relevance. Limitations, reasons for caution The retrospective design of our study may have introduced certain limitations and potential biases. To overcome these limitations, it is advisable that future research endeavors adopt a prospective clinical study design. Wider implications of the findings This study uses a diverse dataset and rigorous methods to create a user-friendly prediction model for early miscarriage in ART. By considering regional differences, it ensures the model’s broader applicability and accuracy, enabling personalized treatment strategies and improving infertility treatment outcomes. Trial registration number The study was a retrospective study, and it was approved by the Institutional Review Boards of Sun Yat-Sen Memorial Hospital (SYSEC-KY-KS-2021-121) and the First Affiliated Hospital of Xinjiang Medical University (K202106-17) and is reported in accordance with the TRIPOD statement

Non-invasive cumulus cell analysis can be applied for oocyte ranking and is useful for countries with legal restrictions on embryo generation or freezing.

Can a strategy for scoring oocyte quality, based on cumulus cell (CC) gene expression, prioritize oocytes with the highest implantation potential, while limiting the number of embryos to be processed in culture and the number of supernumerary embryos to be vitrified? An interventional, blinded, prospective cohort study was retrospectively analyzed. In the original study, patients underwent a fresh Day3 single embryo transfer with embryos ranked based on morphology and CC gene expression (Aurora Test). The additional ranking of the embryos with the Aurora Test resulted in significant higher clinical pregnancy and live birth rates. Now it is investigated if the Aurora Test ranking could be applied to select oocytes. The effect of an Aurora Test based restriction to 2 and 3 2PN or MII oocytes on clinical pregnancy and other outcomes, was analyzed in two subsets of patients with all 2PN (n = 83) or all MII oocytes (n = 45) ranked. Considering only the top three ranked 2PN oocytes, 95% of the patients would have received a fresh SET on Day3 resulting in 65% clinical pregnancies. This was not different from the pregnancy rate obtained in a strategy using all oocytes but significantly reduced the need for vitrification of supernumerary embryos by 3-fold. Considering only top-ranked MII oocytes gave similar results. In countries with legal restrictions on freezing of embryos, gene expression of CC can be used for the selective processing of oocytes and would thus decrease the twin pregnancy rate and workload, especially for embryo morphology scoring and transfers as the handling and processing of lower competence oocytes is prevented, while improving the ART outcome.

Cumulative live birth rate in mild versus conventional stimulation in progestin-primed ovarian stimulation protocols for individuals with low prognosis.

The purpose of this study was to evaluate the cumulative live birth rate (CLBR) of mild stimulation and conventional stimulation for the low-prognosis population undergoing PPOS protocols. This was a retrospective cohort study. We included women with a low prognosis. All women underwent PPOS protocols, and the starting gonadotropin (Gn) dose was 150 IU or 300 IU. The primary outcome measure was CLBR. The secondary outcome measures were the number of oocytes retrieved, number of 2PN oocytes and number of available embryos. In total, 171 women with mild stimulation and 1810 women with conventional stimulation met the criteria. In the PSM model, 171 mild stimulation cycles were matched with 513 conventional stimulation cycles. The gonadotropin dosage in the mild stimulation group was significantly lower than that in the conventional stimulation group (1878.6 ± 1065.7 vs. 2854.7 ± 821.0, P<0.001). The numbers of oocytes retrieved, 2PN oocytes, available embryos and high-quality embryos were also higher in the conventional stimulation group than in the mild stimulation group (P<0.05). There was no significant between-group difference in the cumulative clinical pregnancy rate (26.3% vs. 27.5%, P=0.77). The CLBR after mild stimulation was similar to that after conventional stimulation (21.1% vs. 22.0%, P=0.79). In our study, we found that the CLBRs of mild stimulation and conventional stimulation were similar, despite conventional stimulation resulting in significantly more oocytes and embryos. Thus, mild stimulation can be considered an option for women with a low prognosis in PPOS protocols.

P-030 Clinical outcomes of microdissection testicular sperm extraction (micro TESE) and intracytoplasmic sperm injection (ICSI) in non-obstructive azoospermia (NOA) with the history of cryptorchidism

Abstract Study question What is sperm retrieval rate (SRR) by micro TESE and the clinical outcomes using testicular sperm in couples with the history of cryptorchidism? Summary answer NOA couples with the history of cryptorchidism had a higher SRR by micro TESE but lower clinical outcomes by ICSI compared of unexplained NOA. What is known already Undescended testis (UT) which is exposed to a higher temperature compared with the scrotal temperature is associated with impairment of germ cell maturation, and progressive Leydig and Sertoli cell atrophy, and subsequent infertility in adulthood. There have been very few studies of ICSI with a focus on, or large enough numbers to examine, the specific outcomes associated with male factor infertility, however improvement in sperm retrieval techniques including micro TESE and micromanipulation techniques, such as ICSI, has led to excellent fertilization and pregnancy outcomes of treatment cycles. Study design, size, duration We performed a retrospective study based on two reproduction centers in Japan and evaluated 1521 azoospermic patients in our clinics between September 2013 and December 2022. We investigated SRR by micro TESE in these patients and therefore aimed to evaluate the prevalence and the significance of ICSI outcomes with embryonic development in NOA couples with the history of cryptorchidism. Participants/materials, setting, methods We evaluated SRR of micro TESE, two pronuclei (2PN) oocyte rates, blastocyst development, good-quality blastocyst (Grade 3BB and above on day 5 by the Gardner scoring), and clinical pregnancy rates per embryo transfer (ET) in 72 NOA cases with the history of UT, 953 cases of unexplained NOA, not including after orchidopexy, Klinefelter syndrome, cryptozoospermia, mumps orchitis, and 284 cases of obstructive azoospermia (OA). Statistical analysis was performed using unpaired t-tests and chi-squared tests. Main results and the role of chance SRR of first attempt micro TESE in UT (44/56=78.6%) was higher than unexplained NOA (160/748=21.4%) (p &amp;lt; 0.001). Spermatozoa were successfully retrieved in 13 of 22 (59.1%) UT group and 36 of 226 (15.9%) unexplained NOA who had previously undergone micro TESE with no sperm found. No correlation was found between serum FSH, LH, and T level with the success of sperm retrieval. Testicular volume and patient age at orchidopexy also did not affect the SRR for micro TESE. 2PN oocytes, blastocysts development, and good-quality blastocysts rates were 45.5%, 47.4%, and 18.9% in UT, 51.2%, 45.0%, and 20.9% in unexplained NOA, and 62.6%, 52.1%, and 23.7% in OA, respectively. Clinical pregnancy rates per ET were 29.3% in UT, 32.3% in unexplained NOA, and 41.6% in OA. Significant differences were only observed in 2PN oocytes between unexplained NOA and OA (p &amp;lt; 0.05) and clinical pregnancy between UT and unexplained NOA (p &amp;lt; 0.05), and OA and unexplained NOA (p &amp;lt; 0.01). Several UT patients showed a very small number of spermatozoa and even only immotile sperm of retrieved by micro TESE. Limitations, reasons for caution We included the patients only after a surgery of cryptorchidism, but not a delayed testicular descent without surgery. The cohort size of this study is not small, however, our screened population of azoospermic patients may be biased. Wider implications of the findings This study shows a high impact in micro TESE and ICSI outcomes with embryonic development for the NOA couples with the history of cryptorchidism. Our study emphasizes that history of cryptorchidism provides clinically valuable prognostic information to couples considering surgical sperm retrieval. Trial registration number not applicable

P-633 Progesterone elevation on the day of trigger in the presence of low ovarian response signifies abnormal follicular environment and poorer oocyte health

Abstract Study question Is progesterone elevation (PE) in the presence of low number of follicles (unexpected PE) associated with oocyte or embryo quality? Summary answer PE on day of trigger in cycles with ≤4 follicles is associated with significantly reduced fertilisation rates and fewer top-quality blastocysts What is known already The negative effect of PE on pregnancy rates in fresh IVF cycles has been demonstrated by multiple studies and meta-analyses, and most studies support no negative effect on oocyte and embryo quality. However, as it is thought that PE is the product of cumulative secretion of low levels of progesterone from multiple developing follicles, PE in the presence of a low number of follicles (unexpected PE) is considered a different pathophysiological entity. It has been hypothesized, but never proven, that in these cases, PE is the result of follicular dysregulation and that it might indicate poorer follicular and oocyte health. Study design, size, duration This is a multi-centre retrospective analysis of a large cohort (n = 1,393) of IVF/ICSI cycles with a low response to ovarian stimulation (1-4 follicles ≥14mm) performed from January 2018 to April 2022. Only cycles of ovarian stimulation using gonadotrophins and GnRH analogues with a measurement of serum progesterone on the day of triggering final oocyte maturation were included. Natural cycles and cycles involving the administration of clomiphene were excluded. Participants/materials, setting, methods Unexpected PE was defined as serum progesterone levels greater than 4.77 nmol/L (1.5 ng/mL) on day of trigger. The primary outcome was the number of usable blastocysts. Secondary outcomes included the number of oocytes retrieved, two pronuclei (2PN) oocytes, number of top-quality embryos, fertilisation rate and blastocyst formation rate. Generalised estimating equations were used to account for the clustered nature of data. Multivariable regression analysis was performed to control for the effect of multiple confounders. Main results and the role of chance In the univariate regression analysis, the number of oocytes retrieved was not significantly different between cycles with and without PE (mean 2.3 vs. 2.1 respectively; p = 0.550). The number of 2PN oocytes was significantly (p = 0.021) lower in the PE group (mean 0.72, 95%CI 0.31-1.14) compared to the non-PE group (mean 1.21, 95%CI 1.12-1.31). The number of top-quality blastocysts was also significantly (p = 0.003) lower in the PE group (mean 0.03, 95%CI -0.03-0.10) compared to the non-PE group (mean 0.14, 95%CI 0.11-0.17). Fertilisation rates per oocyte retrieved, fertilisation rates per oocyte inseminated and the top-quality blastocyst rate per oocyte retrieved were consistently lower in the PE group at 33.3% (95% CI 19.0-47.6), 26.9% (95% CI 14.8-39.0) and 0.7% (95% CI -0.8-2.1) compared to 58.7% (95% CI 55.8-61.7), 52.3% (95% CI 49.3-55.2) and 5.5% (95% CI 4.3-6.8) in the non-PE group. The negative association of PE with the primary outcome measure of this study was confirmed in the multivariable regression analysis where the number of usable blastocysts was significantly lower in the PE group (adjusted mean 0.14, 95%CI 0.03-0.25) compared to the non-PE group (adjusted mean 0.27, 95%CI 0.23-0.31) after adjusting for confounders. Limitations, reasons for caution This is a retrospective observational study, thus not all confounders may be accounted for. Despite being one of the largest studies conducted in poor responders, the sample size of unexpected PE cycles is relatively small due to its rarity. Therefore, this study may be underpowered to detect smaller effect sizes. Wider implications of the findings This study demonstrates for the first time that unexpected PE on the day of trigger (i.e. in cycles with low ovarian response) is associated with poorer oocyte health as reflected in the reduced fertilisation rates and fewer blastocysts available for transfer. This information can aid in better understanding this phenomenon. Trial registration number Not applicable

P-669 The effect of ovarian follicle size on oocyte fate and embryology outcome

Abstract Study question Does ovarian follicle size influence ultimate oocyte fate and embryology outcome? Summary answer Per punctured follicle, higher oocyte yield, MII oocytes, 2PN fertilisation, and utilisable blastocyst is observed as follicle increases in size up to threshold of 15.5mm What is known already It is well known that oocyte maturity and probability of successful fertilisation is correlated to the size of follicles where oocyte originated from. However the true cutoff and ideal size at oocyte retrieval remains elusive. Furthermore, there is no conclusive evidence that embryological outcome in terms of blastocyst morphology and utilisation differs in the mid to large ovarian follicular sizes. Study design, size, duration Prospective observational study undertaken in a tertiary fertility referral centre over 10 months. 50 participants recruited and underwent conventional controlled ovarian stimulation as per unit protocol. A total of 288 follicular punctures performed in the study period. Participants/materials, setting, methods 50 oocyte retrieval (OR) procedures were performed. The size of ovarian follicles was individually measured using transvaginal ultrasound. 288 follicular punctures were performed and up to 3 oocytes per patient were collected for the study to be individually cultured to allow for tracking of oocyte and embryo outcomes. Follicular fluid and spent culture media were collected separately for subsequent metabolomics analysis. Follicular sizes were divided into 6 groups ( &amp;lt; =9.5, 10-12.5, 13-15.5, 16-19.5, 20-23.5, &amp;gt; =24). Main results and the role of chance Oocyte fate included 113 mature oocytes, 83 bipronuclear (2PN) oocytes, 7 cleavage stage embryos, and 39 good quality blastocysts suitable for utilisation in fresh or frozen transfer cycles. When compared to overall average per punctured follicle, there is an association of higher oocyte yield, proportion of MII oocytes, 2PN fertilisation, and utilisable blastocyst as follicle size increases. The threshold for a change in fate appears to be when follicle size increases beyond 15.5mm where optimal follicle size may reach plateau. Proportion of good-quality blastocysts appears to be associated with size of follicles at rates as follow- 0% (less than 10mm), 11.1% (10-12.5mm), 13.2% (13-15.5mm), 15.9% (16-19.5mm), 14.5% (20-23.5mm), and 20% (above 24mm). 23 embryos have been utilised to date, yielding a 50% positive pregnancy rate. Of these utilised embryos resulting in a positive pregnancy test, 81.8% of embryos originated from follicles above 15.5mm. Metabolomics analysis for follicular fluid and spent culture media is ongoing. Limitations, reasons for caution The observational nature of the study along with a small sample size are limitations that did not allow clinical significance to be observed. Wider implications of the findings Oocyte retrieved from follicles less than 10mm is futile. There appears to be a plateau in optimal follicle size where no real detriment is demonstrated. This is an important findings to guide stimulation, especially in cases of asynchronous growth where sacrifice in larger follicles is commonly assumed. Trial registration number Not applicable

P-251 Cumulus cell analysis as a non-invasive oocyte selection strategy to reduce the number of oocytes/embryos cultured and increase pregnancy rates

Abstract Study question Can non-invasive gene expression analysis of cumulus cells (CC) improve efficiency in ART by prioritizing oocytes for further culture and fresh single embryo transfer? Summary answer CC analysis can be used for the selective processing of oocytes. This may reduce culture work and improve the outcome in ICSI elective SETs (eSET). What is known already In an interventional, blinded, prospective cohort study (Van Vaerenbergh et al. 2021), 113 patients underwent a fresh Day3 eSET with embryos ranked and transferred based on morphology and CC gene expression (Aurora Test), while 520 control patients underwent a Day3 eSET without the Aurora Test. This resulted in a significant higher clinical pregnancy of 61% in the patients with eSET based on CC ranking applied on good morphology embryos, compared to 29% in the controls with eSET based on embryo morphology only. Live birth rate was also significantly increased, while time-to-pregnancy was significantly reduced with 3 transfer cycles. Study design, size, duration In a retrospective analysis, in a subset of patients with at least 6 growing follicles and at least five 2PN oocytes (n = 80), it was investigated whether the Aurora Test, used to select transferrable Day3-embryos, could also be applied to select oocytes on Day0/1. The effect of processing only the three highest ranked oocytes (based on the Aurora Test) on embryo development and clinical pregnancy was studied compared to processing all oocytes. Participants/materials, setting, methods Patients included in this single centre study had their first or second GnRH-antagonist ICSI cycle, were younger than 40y, had normal BMI, were stimulated with HP-hMG and scheduled for Day3 eSET. Two-sided statistical analysis (p &amp;lt; 0,05) was performed between a strategy of processing only the top 3 Aurora ranked oocytes, according to CC gene expression, and a strategy of processing all available oocytes. Main results and the role of chance On average, 8 MII oocytes were obtained per patient and the average fertilization rate was 83%. In total, 407 good quality embryos (GQE) on Day3 were generated from these 80 patients when utilising all 639 oocytes. Processing the three top-ranked oocytes only (240/639 oocytes) would have reduced the number of embryos to 169 GQE and would have resulted in 2.1 GQE on average on Day3 per patient; 75/80 (94%) patients would have had a fresh Day3 transfer resulting in a 63% clinical pregnancy rate. Processing all 639 available 2PN oocytes (standard of care) resulted in a fresh Day3 transfer in all 80 patients and a similar 64% clinical pregnancy rate (ns). However, 399 more oocytes would need to be processed. The strategy of restricting the number of oocytes to be processed would not have compromised cumulative cycle outcome. Considering all subsequent freeze/thawing cycles the cumulative clinical pregnancy rate calculated per all 80 patients would increase to 90%. Limitations, reasons for caution The limitation of this approach is that the Aurora Test requires individual oocyte denudation and individual oocyte vitrification. Secondly, this new strategy should be validated in a prospective study. Wider implications of the findings By applying this oocyte selection strategy patients would benefit from a high pregnancy rate in the fresh transfer cycle, while the lab would see reduction in embryo culture work, because freeze/thawing cycles and culture of embryos with lower competence would be prevented. Trial registration number NA

P-217 Laboratory and clinical outcomes for fresh and vitrified-warmed oocytes after Piezo-intracytoplasmic sperm injection (Piezo-ICSI)

Abstract Study question How is the performance of Piezo-ICSI on vitrified-warmed oocytes compared to fresh oocytes in human IVF? Summary answer Laboratory outcomes for fresh and vitrified-warmed oocytes after Piezo-ICSI were comparable. What is known already ICSI is utilized as the fertilization method for 60-80% IVF patients undergoing assisted reproductive technology (ART) worldwide. In the past 3 decades, Piezo-ICSI (Prime Tech Ltd, Japan), a refined ICSI methodology had shown increased fertilization and survival rates. The fertilization rate (2PN, 1PN, and &amp;gt;2PN) achieved via Piezo-ICSI was reported as high as 84-91% in previous publications. Study design, size, duration This retrospective cohort study included 12,195 fresh and 1,065 vitrified-warmed oocytes retrieved or warmed from 1334 and 93 patients respectively between January 2020 and November 2021. Mean age was 35.9 (ranged 19-49) for fresh oocytes and 27.3 (ranged 18-49) for vitrified-warmed oocytes. The vitrified-warmed oocytes were mainly originated from young donors and thus reflected in lower maternal age in this cohort Participants/materials, setting, methods All oocytes fertilized using Piezo-ICSI technology during the study period were included. Piezo-ICSI is performed using a piezo impact drive unit (MB-S; Prime Tech Ltd, Japan) driven by a controller (PMM4G; Prime Tech Ltd, Japan). The ultrathin walled micropipette (PINU06-25FT; Prime Tech Ltd, Japan) filled with 1-1.5cm operation liquid (Flourinert FC-770; Sigma, Belgium) was used for microinjection. For vitrified-warmed oocytes, Cryotec Method (Reprolife, Japan) was used for both vitrification and warming of oocytes. Main results and the role of chance The two groups (fresh and vitrified-warmed oocytes) were comparable (p &amp;gt; 0.05) in terms of fertilization (2PN and &amp;gt;2PN) rate (81.5% vs 80.0%), post-ICSI oocytes survival rate (93.3% vs 92.8), blastocyst-formation rate from 2PN oocytes (74.6% vs 71.0%) and blastocyst-utilization rate from 2PN oocytes (53.2% vs 49.3%). This study demonstrated that fresh and vitrified-warmed oocytes achieved comparable laboratory outcome following the use of Piezo-ICSI technique and ultrathin walled injection pipette. Limitations, reasons for caution The methodology in performing Piezo-ICSI differs compared to conventional ICSI, hence training is required to achieve optimal results. Wider implications of the findings Our data resonates with previously published data using the same Piezo-ICSI platform. Trial registration number Not applicable

P-613 Association between basal androgen concentrations and number of follicles on the day of triggering final oocyte maturation in poor responders undergoing IVF; A prospective study

Abstract Study question Are basal androgen concentrations associated with the number of follicles on the day of triggering final oocyte maturation in poor responders undergoing in vitro fertilization (IVF)? Summary answer A significant negative association is present between (dehydroepiandrosterone sulfate) DHEAS concentrations and the number of follicles on the day of triggering final oocyte maturation. What is known already Studies in animals have shown that androgens promote early follicular development and granulosa cell proliferation, by augmenting follicle-stimulating hormone (FSH) receptor expression in granulosa cells. Several retrospective studies have evaluated the association between basal androgen concentrations and follicular development in normal and poor responders undergoing ovarian stimulation for IVF with conflicting results. Study design, size, duration This prospective study was performed between 02/2020 and 01/2022 in 103 poor responders according to the Bologna criteria. Androgens, including total testosterone, sex hormone-binding globulin (SHBG), dehydroepiandrosterone sulfate (DHEAS), Δ4-androstenedione and 17-OH progesterone (17-OHP), were measured at the initiation of ovarian stimulation, using the automated Elecsys immunoanalyser (Roche Diagnostics, Mannheim, Germany). Ovarian stimulation was performed using a fixed dose of 300 IU of recombinant gonadotrophins and gonadotrophin-releasing hormone (GnRH) analogues. Participants/materials, setting, methods Triggering of final oocyte maturation was performed in the presence of three follicles of ≥ 17mm. The primary outcome measure was the number of follicles ≥11mm on the day of triggering final oocyte maturation. The association between androgen concentrations and the number of follicles ≥11mm on the day of triggering was evaluated using generalized estimating equations, accounting for female age and body mass index (BMI). Values were expressed as coefficient (coef) or mean (95% confidence interval). Main results and the role of chance Female age was 41.9 (41.2-42.6) years, while BMI was 26.1 (24.9-27.3) kg/m2. The duration of ovarian stimulation was 10.3 (9.7-10.8) days and the total dose of gonadotrophins required was 3058 (2903-3213) IU. The number of follicles ≥11 mm on the day of triggering final oocyte maturation was 6.1 (5.3-7.0). The number of COCs retrieved was 3.9 (3.2-4.6), the number of MII oocytes was 3.4 (2.8-3.9) and the number of 2pn oocytes was 2.5 (2.1-2.8). No significant association was found between basal testosterone (coef: -0.008, -0.019 to + 0.003, p = 0.17), 17-OHP (coef: -0.044, -0.391 to + 0.303, p = 0.80), SHBG (coef: -0.002, -0.007 to + 0.002, p = 0.25) Δ4-androstenedione (coef: -0.101, -0.306 to + 0.104, p = 0.33) concentrations and the number of follicles ≥11mm on the day of triggering final oocyte maturation. In contrast, a significant negative association was found between basal DHEAS (coef: -0.011, -0.019 to -0.003, p = 0.007) concentrations and the number of follicles ≥11 mm on the day of triggering final oocyte maturation. Higher DHEAS concentrations were associated with the development of fewer follicles ≥11 mm. Limitations, reasons for caution This prospective study evaluated the association between basal androgen concentrations and the number of follicles on the day of triggering final oocyte maturation in poor responders stimulated with a fixed dose of recombinant FSH. However, the precision of the estimates could be increased by analyzing a larger study population. Wider implications of the findings DHEA supplementation in poor responders undergoing IVF has not been shown to improve ovarian response. Given the significant negative association between DHEAS concentrations and the number of follicles on the day of triggering final oocyte maturation, future studies on DHEA supplementation should consider basal DHEAS concentrations. Trial registration number N/A

O-127 Higher probability of live birth after testosterone pretreatment in women with poor ovarian response undergoing IVF: a systematic review and meta-analysis

Abstract Study question Does testosterone pretreatment increase the probability of pregnancy in poor responders undergoing ovarian stimulation with gonadotrophin-releasing hormone (GnRH) analogues and gonadotrophins for in-vitro fertilization (IVF)? Summary answer Testosterone pretreatment increases clinical pregnancy and live birth rates in poor responders undergoing ovarian stimulation for IVF. What is known already Androgens have been shown to stimulate early stages of follicular growth, increase the number of primary, pre-antral and antral follicles as well as increase ovarian sensitivity to follicle stimulating hormone (FSH). Although androgen supplementation has been evaluated in several randomized controlled trials (RCTs) and meta-analyzed in six systematic reviews until today currently no solid conclusions can be drawn regarding its effectiveness. Study design, size, duration A literature search was performed until September 2021 aiming to identify RCTs evaluating testosterone pretreatment in poor responders. Outcome measures included achievement of pregnancy, total dose of gonadotrophins required, duration of stimulation, estradiol levels, endometrial thickness and number of follicles ≥17 mm on the day of triggering final oocyte maturation, number of cumulus–oocyte complexes (COCs) retrieved, embryos transferred, metaphase II (MII) and 2-pronuclei oocytes (2pn) and the proportion of patients having an embryo transfer (ET). Participants/materials, setting, methods Eight RCTs published between 2006 and 2021 were analyzed, including 760 women. Pretreatment with transdermal testosterone gel was performed in all studies with a dose ranging from 10 to 12.5 mg/day for 10 to 56 days. In dichotomous data, estimates were expressed as risk ratio (RR) with 95% confidence intervals (CIs), using the fixed or random effects method. In continuous data, differences were pooled across resulting in a weighted mean difference (WMD) with 95% CI. Main results and the role of chance Testosterone pretreatment was associated with a significantly higher live birth (RR: 2.07, 95%CI: 1.09 to 3.92) and clinical pregnancy rate (RR: 2.25, 95%CI: 1.54) in women with POR undergoing IVF, while there was also a significant increase in the number of COCs retrieved (WMD: +0.88, 95% CI: +0.22 to + 1.54). Significantly less days to complete ovarian stimulation (WMD: -0.81 days, 95% CI: -1.46 to −0.16), a lower total dose of gonadotrophins (WMD: -368.8 IUs, 95% CI: −612.4 to -125.2), a thicker endometrium on the day of triggering final oocyte maturation (WMD: +0.83 mm, 95% CI: +0.13 to + 1.53) and a lower cancellation rate due to poor ovarian response (RR: 0.37, 95%CI: 0.20 to 0.71) were observed. No significant differences were observed in estradiol levels (WMD: -8.12 pg/mL, 95% CI: -118.2 to + 101.96), in the numbers of follicles ≥17 mm on the day of triggering final oocyte maturation (WMD: +0.82, 95%CI: -0.11 to + 1.74), of MII oocytes (WMD: +0.50, 95% CI: -0.17 to + 1.17), of 2pn oocytes (WMD: +0.49, 95% CI: -0.11 to + 1.10), of embryos transferred (WMD: +0.21, 95%CI: -0.07 to + 0.49) and in the proportion of patients with ET (RR: 1.00, 95% CI: 0.96 to 1.04). Limitations, reasons for caution The definition of poor ovarian response varied among studies and a considerable heterogeneity regarding the type, dose and duration of testosterone pretreatment was present. Although the present study is currently the largest meta-analysis evaluating testosterone pretreatment, the total number of patients is still not large enough to draw solid conclusions. Wider implications of the findings The current study suggests that the probability of pregnancy is increased in poor responders pretreated with transdermal testosterone. This increase, in the absence of other proven beneficial interventions in these patients, justifies the conduction of further relevant RCTs. Trial registration number CRD42021262098

Fertilization and neonatal outcomes after early rescue intracytoplasmic sperm injection: a retrospective analysis of 16,769 patients

PurposeTo evaluate the efficacy and safety of short-term insemination and early-rescue intracytoplasmic sperm injection (ICSI), an approach that rescued oocytes with unclear second polar body 6 h after initial insemination by ICSI (early R-ICSI) to avoid total or near-total fertilization failure in conventional in vitro fertilization (IVF).MethodsWe performed a retrospective study in 16,769 patients (short-term IVF, n = 12,094; ICSI, n = 3452; early R-ICSI, n = 1223) who received IVF/ICSI treatment in our hospital from January 2009 to October 2018. Fertilization and clinical outcomes were compared among those three groups.ResultsWhen considering the R-ICSI embryos in the early R-ICSI group independently, the rates of fertilization and day-3 cleaved embryos in 2PN oocytes were comparable, the rates of fertilization (2PN) and high-quality embryos were lower, whereas the multi-PN fertilization rate (3.27%) was significantly higher than the ICSI group (1.26%). The difference of clinical pregnancy rate between the part of transferred R-ICSI embryos (40.81%) and the ICSI group (44.73%) remained nonsignificant. Furthermore, the rate of congenital birth defects in the early R-ICSI group (0.99%) was not significantly different from those in the short-term IVF (0.76%) and ICSI groups (1.07%).ConclusionDespite the multi-PN fertilization rate, our study highlights early R-ICSI as a safe and effective alternative in assisted reproduction to decrease complete IVF fertilization failure and reduce ICSI utilization. Additional large amount and long-term follow-up studies are needed to further validate the use of early R-ICSI.

The effect of ovarian follicle size on oocyte and embryology outcomes

To identify relationships between the size of punctured ovarian follicles and subsequent embryology outcomes. Prospective observational cohort study. Private fertility center. One hundred fifty-seven oocyte retrievals performed during the study period. The diameter of punctured follicles was ultrasonically measured during routine oocyte collection. The resulting embryos were group-cultured to the blastocyst stage and classified into 8 groups according to follicle size (≤9.5, 10-12.5, 13-15.5, 16-18.5, 19-21.5, 22-24.5, 25-27.5, and ≥28 mm). Rate of good-quality blastocysts per follicle puncture. This study included 4,539 follicle punctures, 2,348 oocytes, 1,772 mature oocytes, 1,258 bipronuclear (2pn) oocytes, and 571 good-quality blastocysts derived from 157 oocyte retrievals. The per-puncture yields of oocytes, mature oocytes, 2pn oocytes, and good-quality blastocysts were associated with the size of the punctured follicle. The rates of good-quality blastocysts per punctured follicle were 2.2% (≤9.5 mm), 6.2% (10-12.5 mm), 11.9% (13-15.5 mm), 14.5% (16-18.5 mm), 18.9% (19-21.5 mm), 17.5% (22-24.5 mm), 15.9% (25-27.5 mm), and 16.0% (≥28 mm). When compared with the overall average, punctures of follicles in groups ≤12.5 mm in diameter had significantly inferior yields of good-quality blastocysts, whereas punctures of follicles in groups 19-24.5 mm in diameter were associated with significantly greater than average yields of good-quality blastocysts. Other groups did not differ significantly from average. No correlation was observed between follicle diameter and ploidy of biopsied blastocysts. Punctures of follicles ≤12.5 mm in diameter rarely result in good-quality blastocysts. The yield of good-quality blastocysts progressively increases with follicle size up to approximately 19 mm in diameter, with no substantial decline above that size. The ploidy of the blastocysts that form appears to be unaffected by follicle size.

THE PRE-IMPLANTATION HUMAN EMBRYO “REMEMBERS” THE SIZE OF THE OVARIAN FOLLICLE THAT IT CAME FROM

Investigate any relationship between ovarian follicle size and the rate of good blastocyst formation among successfully fertilized (bipronuclear, or 2pn) oocytes. This was an IRB-approved prospective observational study. Following conventional ovarian stimulation with exogenous gonadotropins, follicles were measured during oocyte collection. After insemination by intracytoplasmic sperm injection, oocytes were subsequently observed to assess fertilization. Resulting embryos were group-cultured to the blastocyst stage according to follicle size. Good blastocysts were those suitable for transfer (no “CC” grades by the Gardner system). Logistic regression was used to assess oocyte maturity, fertilization potential, and good blastocyst formation as a function of follicle size. The results of 2152 collected oocytes, 1633 M-II oocytes, 1126 2pn oocytes, and 566 good-quality blastocysts are shown in Table 1. Logistic regression of oocyte maturity versus follicle size found increasing probability of oocytes being mature (metaphase-II) as follicle size increased (P<0.0001). Logistic regression of successful fertilization (2pn) among mature oocytes did not find a significant relationship with follicle size (P=0.1792). Logistic regression of good blastocyst formation among all oocytes collected found the probability of blastocyst formation significantly increased as follicle size increased (P<0.0001). Logistic regression of good blastocyst formation among 2pn oocytes found the probability of obtaining a good quality blastocyst significantly increased as follicle size increased (P<0.0001). Oocyte maturity and good blastocyst formation have positive correlations with ovarian follicle size.

NEONATAL FOLLOW-UP OF BABY BORN DERIVED FROM A TWO PRONUCLEI PLUS ONE MICRO PN OOCYTE FOLLOWING INTRACYTOPLASMIC SPERM INJECTION

A micro PN is very small and can be overlooked when observed under a microscope. Furthermore, it is difficult to see when it overlaps with the normal nucleus. Out of 5,026 oocytes derived by intracytoplasmic sperm injection, 0.7% showed micro PN. After performing chromosome and ploidy analysis on embryos that have grown to blastocysts, micro PN-derived balanced-diploid blastocysts have been transferred into three patients and childbirth has been reported in two patients. (Capalbo et al, Fertility and Sterility, 2017) However, there are no reported cases of developmental follow-up of babies born from micro PN oocytes. Therefore, we report on the neonatal follow-up of baby born derived from a two pronuclei plus one micro PN oocyte. Patient seeking infertility treatment in a well-established private IVF clinic. The birth weight and the height and weight at one and a half and three years old of one case in which a micro-PN-derived blastocyst was transferred were compared with the results of the 2010 Infant Physical Growth Survey by the Ministry of Health, Labor and Welfare (MHLW) of Japan. The patient was a nulliparous woman and treatment started at the age of 31. Therapy started in August 2009, and AIH was performed 6 times in total upto February 2011, but pregnancy was unsuccessful. After that, 19 oocytes were collected by controlled ovarian stimulation in April 2014. As a result of insemination of the collected oocytes, microPN (size: 9 μm) appeared in one of the ICSI-derived embryos, which was about one-third the size of the male and female pronuclei. Afterwards singleton vitrified-warmed embryo transfer in hormone replacement cycle was performed twice by July 2014. As a result of transferring micro PN-derived blastocyst (Gardner’s score 3AB) at the time of the second embryo transfer, a girl was born by Caesarean section at 38 weeks, and the weight was 3.02 kg, which was within the normal range of the growth curve of data from children in MHLW database. The height at one and a half years was 74.8 cm and the weight was 9.45 kg, which was within the normal range of the growth curve of the children by MHLW. At the age of three, she was 87 cm tall and weighed 13 kg. The height was slightly below the growth curve, but there were no other abnormalities in the child. Follow up from birth to age 3 years did not report any abnormalities or severe growth retardation in the infant.

P–146 Differential impact of three embryo culture media for IVF on in vitro development and perinatal outcome: a single-center RCT

Abstract Study question This report provides updated data from an RCT determining which embryo culture medium yields optimal IVF outcomes. Summary answer Embryo culture systems used for IVF differentially affected preimplantation development and resultant obstetric and perinatal outcomes, including birthweights of live-born singletons. What is known already Currently, multiple embryo culture medium systems are in use for IVF, raising questions regarding which is optimal. However, the ability of a medium to yield preimplantation embryos is not necessarily indicative of embryo viability. For example, supplementation of medium with serum was commonly used to increase animal blastocyst yields, but this impaired embryonic, fetal, and offspring health. In humans, medium composition and culture duration can influence IVF efficacy and offspring phenotype. Given the importance of culture systems in determining clinical outcomes, existing data regarding differential culture system impacts are insufficient and additional well-designed studies are required. Study design, size, duration Between February 2016 and August 2017, 795 couples undergoing their first autologous clinical IVF cycle and freeze-all strategy were recruited. Participants were randomized via computer-generated tables into three groups. Following standard oocyte retrieval and IVF/ICSI procedures, embryos were cultured using three different culture media, G1 Plus/G2 Plus (G1/G2; Vitrolife), Global Total (GT; LifeGlobal), or Sequential Cleav/Sequential Blast (SC/SB; Origio). Thirty-eight patients exhibiting no 2PN oocytes following insemination or those undergoing fresh embryo transfers were excluded. Participants/materials, setting, methods For patients yielding a single good-quality cleavage-stage (day–2 or day–3) embryo, that cleavage-stage embryo was vitrified. For patients yielding two or more good-quality cleavage-stage embryos, two or less good-quality cleavage-stage embryos were vitrified. The culture period of the remaining embryos was extended, and all good-quality blastocyst-stage (day–5 or day–6) embryos were vitrified. This report presents data for vitrified embryo transfer performed until the end of December 2020. Main results and the role of chance The mean per-cycle vitrified embryo yield (± SD) was comparable between groups for cleavage-stage embryos, but significantly different for blastocyst-stage embryos (G1/G2: 1.69 ± 2.2, GT: 2.53 ± 3.01, SC/SB: 2.04 ± 2.42; P = 0.001). Following vitrified cleavage- or blastocyst-stage embryo transfers, biochemical pregnancy rates were significantly different between groups (G1/G2: 55.6%, GT: 59.1%, SC/SB: 46.2%; P = 0.011). Furthermore, a between-group trend towards different live birth rates was observed (G1/G2: 41.7%, GT: 42.1%, SC/SB: 33.1%; P = 0.063). Of 382 live births, data for first-borns (n = 323; 295 singletons and 14 twin-pairs) are reported here. Perinatal data did not differ significantly between groups for both cleavage- and blastocyst-stage embryo transfers, including gestational age- and gender-adjusted singleton birthweight (z-score). Following multiple linear regression (including selected covariates), adjusted mean singleton birthweights were significantly lower in the G1/G2 and GT groups than in the SC/SB group (by 131 g; P = 0.011 and 110 g; P = 0.032, respectively) and tended to be lower for cleavage-stage embryo transfers than for blastocyst-stage embryo transfers (by 102 g; P = 0.053). Limitations, reasons for caution A larger cohort size and longer-term follow-up are required to verify and further elucidate the impact of embryo culture methods on child health. Such studies will raise awareness regarding the sensitivity of in vitro-cultured human embryos to their environment, ultimately resulting in practices that decrease IVF risks to offspring. Wider implications of the findings: Pregnancy outcome of the medium yielding fewer blastocysts was comparable or superior to that of other media, highlighting the importance of differentiating between the ability to support preimplantation development versus the ability to yield viable embryos. Embryo culture medium had a greater impact than embryo transfer stage on live birthweight. Trial registration number UMIN000020910

P–019 Sperm parameter and ICSI / IVF outcomes after sperm selection using microfluidic sperm separator and density gradient centrifugation with swim-up in split semen sample

Abstract Study question To analyze whether microfluidic sperm selection (MSS) by ZyMōt™ improves sperm DNA fragmentation rate and embryonic development compared to density gradient centrifugation with swim-up (DGCS). Summary answer MSS by ZyMōt™ selects sperm for clinical use with less DNA damage significantly compared to DGCS. What is known already Conventional sperm preparation methods, such as density gradient centrifugation and the swim-up method utilize centrifugation during processing, may damage the sperm. MSS may allow for improved selection of normal sperm compared with conventional sperm preparation as it yields sperm with a lower DNA fragmentation rate. However, there are few clinical studies by sibling oocytes study compared to DGCS. Study design, size, duration This prospective study was performed between March 2020 and May 2020 at a reproductive center. All patients involved gave written consent, and institutional review board approval was granted. A total of 575 metaphase II oocytes were collected from 49 cycles. Wife’s age was 34.7 ± 3.9 years old. Raw sperm concentration and motile sperm concentration was 63.1 ± 78.7M/mL, and 41.6 ± 67.7M/mL, respectively. Participants/materials, setting, methods Patients who performed ART for the first or second time were divided into two groups according to MSS and DGCS. Sperm DNA fragmentation rate (SDFR) and motile sperm concentration were compered between MSS and DGCS. SDFR was measured by sperm chromatin structure assay (SCSA) using a flow cytometer. Sibling oocytes were randomized into MSS-IVF, DGCS-IVF, MSS-ICSI, and DGCS-ICSI. Rate of two pronuclear (2PN) oocytes, blastocysts development, and good-quality blastocysts were compared between each group. Main results and the role of chance SDFR was 13.5 ± 11.8% for raw semen. SDFR was significantly lower after MSS (3.6 ± 4.1%) than that for raw semen and after DGCS (17.4 ± 14.8%) (P &amp;lt; 0.01). Motile sperm concentration after MSS (19.0 ± 28.3M/mL) was significantly higher after than after DGCS (15.4 ± 15.3M/mL) (P &amp;lt; 0.01). The number of IVF performed was 145 for MSS and 132 for DGCS. IVF results (MSS vs DGCS) were 2PN rate (73.1% vs 72.0%), blastocysts development rate (65.3% vs 55.4%), and good quality blastocysts rate (43.2% vs 34.9%). The number of ICSI performed was 149 for MSS and 149 for DGCS. ICSI results (MSS vs DGCS) were 2PN rate (77.9% vs 79.2%), blastocysts development rate (68.8% vs 65.8%), and good quality blastocysts rate (35.8% vs 30.6%). No significant difference was observed between MSS and DGCS for each parameter both IVF and ICSI. Limitations, reasons for caution The participants were limited to those who collected semen of 2mL or more and motile sperm concentration of above 1M/mL, because semen sample needed to be divided to MSS and DGCS. Wider implications of the findings: This is the first study to conducted in sibling oosytes study with MSS and DGCS, in both IVF and ICSI. MSS is effective in collecting sperm with less DNA damage compared to DGCS. Motile sperm concentration after using MSS is sufficient to perform IVF as well as DGCS. Trial registration number Not applicable

P–259 Blastocyst quality is associated with both the start and the duration of compaction after ICSI

Abstract Study question Is the start and the total duration of compaction related to embryo quality? Summary answer The timing of the start, the end and the total duration of compaction are associated with blastocyst quality grade in the IVF laboratory. What is known already Preimplantation embryo development follows a programmed timeline during which a series of critical events take place. One event typically occurring on day 3/4 post fertilisation is the formation of adherence junctions between blastomeres in a process called compaction. It is considered the first morphological event in the differentiation process of the mammalian embryo. Evaluation of developmental events are used to optimize the selection of the most competent embryos for transfer and/or cryopreservation in the IVF laboratory. It has already been shown that the time of full compaction is indicative for high-quality blastocysts with a higher implantation rate. Study design, size, duration A single-centre retrospective observational study including 74 ICSI cycles performed in 2020. Injected oocytes were cultured in blastocyst medium (Origio) in the EmbryoScope + (Vitrolife) for 5/6 days. Embryos that reached the blastocyst stage were evaluated for the start of compaction, the time to reach full compaction and the total duration of compaction. These parameters were compared between good- and poor-quality blastocysts; the primary outcome parameter of the study was embryo quality. Participants/materials, setting, methods Only ICSI cycles with ejaculated fresh/frozen-thawed sperm and monitored in time-lapse incubator were included. All MNC, IVM and PGT cycles were excluded. Time zero was the start of ICSI. Good-quality embryos were full and expanded blastocysts with good-quality inner cell mass and trophectoderm (AA, AB, BA and BB according to Gardner and Schoolcraft (1999)). GraphPad Prism was used for statistical analysis. After testing for normality and homogeneity, unpaired t-test or Mann-Whitney test determined significant differences. Main results and the role of chance In this study, of the 528 included 2PN oocytes, 229 (43.4%) reached the blastocyst stage and 299 (56.6%) were arrested. Among the former, 131 (57.2%) blastocysts were classified in the good-quality group and 98 (42.8%) blastocysts in the poor-quality group. In general, human embryos compacted slowly while dividing further and the blastomeres moved during the compaction process. The start of compaction was heterogeneous (between 50.9 and 102.7 hours post ICSI; mean=80.0 hours), as well as the cell number at the initiation (between 4 and 18 blastomeres; mean=12 blastomeres). The time analysis showed that the embryos in the good-quality group started to compact significantly earlier than those in the poor-quality group (mean=78.6 vs 82.2 hours; R²=0.06; p &amp;lt; 0.01). We confirmed that blastocysts in the good-quality group reached full compaction earlier than those in the poor-quality group (mean=86.8 vs 93.8 hours; R²=0.17; p &amp;lt; 0.01). Furthermore, the total duration of compaction was significantly lower in the good-quality than in the poor-quality group (median=7.4 vs 10.7 hours; p &amp;lt; 0.01). Limitations, reasons for caution As this is a retrospective study, the influence of uncontrolled variables cannot be excluded. The absence of the pregnancy outcome and live birth rate is a shortcoming and will be subject of a larger patient-to-patient study. Wider implications of the findings: These results indicate that an earlier start and a shorter duration of compaction are associated with better blastocyst quality. These morphological events can be valuable additional parameters in selecting the embryo of better quality when using a time-lapse incubator. Trial registration number Not applicable

AGE-SPECIFIC BLASTULATION RATES IN EMBRYO CRYOPRESERVATION CYCLES YIELDING A CRYOPRESERVED BLASTOCYST

AGE-SPECIFIC BLASTULATION RATES IN EMBRYO CRYOPRESERVATION CYCLES YIELDING A CRYOPRESERVED BLASTOCYST

Associations of FSHR and LHCGR gene variants with ovarian reserve and clinical pregnancy rates

Research questionAre there any associations between the variants of FSHR c.2039 G>A (p. Ser680Asn, rs6166) and LHCGR c.935A>G (p. Asn312Ser, rs2293275) and ovarian reserve, ovarian response, clinical pregnancy rate and POSEIDON group? DesignA total of 210 infertile women were enrolled in this prospective study. The gene variants were analysed by the Sanger method. The clinical parameters were analysed based on genotypes. ResultsThe frequency of heterozygous and homozygous G allele for FSHR c.2039 G>A in the low prognosis group was significantly higher than that in other response groups (P = 0.034); there was no significant association between LHCGR c.935 A>G and ovarian response. Moreover, the serum anti-Müllerian hormone (AMH) concentration, antral follicle count (AFC), oocytes retrieved, metaphase II (MII) oocytes and two-pronuclear (2PN) oocytes in patients with AG genotype for FSHR c.2039 G>A were significantly lower than those with AA genotype. The serum LH concentrations and clinical pregnancy rate of fresh embryo transfer in patients with GG genotype for LHCGR c.935 A>G were significantly higher than that of the AG genotype. In POSEIDON analysis, the low prognosis women with AA genotype for FSHR c.2039 G>A were more likely to appear in subgroup 1 (P = 0.038). ConclusionThe FSHR c.2039 G>A variant has a significant beneficial influence on ovarian reserve and ovarian response. The LHCGR c.935 A>G variant is associated with increased clinical pregnancy rate of fresh embryo transfer in infertile women. In addition, the low prognosis women with AA genotype for FSHR c.2039 G>A tend to show better ovarian reserve and prognosis.