Mean Number Of Embryos Research Articles

The Covid-19 Infection Effect on Assisted Reproductive Method Outcomes.

COVID-19 can have potential pathogenic effects on the oocyte and embryos, but there is limited data about its impact. This study aimed to investigate the COVID-19 impact on the outcome of Assisted Reproduction Techniques (ART) methods. This case-control study was conducted on 190 infertile women who underwent oocyte retrieval at Yas Hospital in vitro fertilization (IVF) department affiliated to Tehran University of Medical Sciences, from October 2021 to October 2022. The case group was defined as women whose PCR test was positive on puncture day and the control group was women with COVID-19 negative tests on puncture day. The study outcome measurements included the number of oocytes retrieved and the number and quality of embryos. Finally, the data were analyzed by SPSS 24. The mean age of the participants was 32.89 ± 5.58 years with an age range of 18-49 years. No significant difference was observed between the two groups regarding baseline variables. The mean number of oocytes was significantly (p =0.001) lower in the case (6.68±4.25) group versus the control (9.07±4.10) group. While there was no statistically significant difference regarding the mean number of embryos in the study groups, No grade A embryos were observed in more than half (57.5%) of the women in the case group. Furthermore, the frequency of grade C embryos on average was 1.08±1.11 in the case group and 0.57±0.75 in the control group, with a statistically significant difference (p =0.010). The findings of this research highlighted that women infected with COVID-19 on the puncture day have a lower number of oocytes and also good-quality embryos.

#371 : Immunological Analysis of Endometrial Receptivity: A New Hope for Patients with Recurrent Implantation Failures

Background and Aim: Recurrent implantation failures (RIF) remain always a strong problem for all IVF teams. Our ART Center started in 2015 and quickly we wanted to focus on implantation failure. After 2 or 3 embryo transfers without pregnancy, guidelines provide to perform exams such as karyotypes, Anti phospholipid, thrombophilia, uterine Doppler, hysteroscopy, endometrial biopsy with CD 136, laparoscopy, sperm DNA Fragmentation. However, a large part of these failures remains unexplained. Following the studies performed in France by N. Lédée in 2013, this study evaluates the results of a new exploration based on immunological analysis of endometrial receptivity. Methods: This study is proposed, after other explorations already done, for patients with 6 or more embryos D3 or 3 embryos D5 transferred without pregnancy. Such an analysis is achieved on endometrial biopsy performed in middle luteal phase and sent to MatriceLab (Paris) Immunological exploration is based on uNK count (uterine Natural Killer cells) and cytokines expression. According to these parameters, “individualized” care is proposed to patient for next embryo transfers to correct the disorder identified by analysis. Results: From 2016 to 2020, 114 biopsies have been performed and sent to MatriceLab. 112 were suitable for analysis. According with results, 4 main profiles have been isolated: Normal immunity (22,3%) Dysregulation immunity (77,7%). Low local immune activation (25%). Over local immune activation (40,2%). “Mix” profile (12,5%) 95 patients have been performing FET following the analysis. Mean age 32.6 ±3.6 years. Mean number of transfers before biopsy 3.9 ±1.6. Mean number of embryos previously transferred 9.6 ±4.4. Deliveries in normal immunity group: 33% (n=6). Deliveries in dysregulation group treated with adapted protocol: 53,2% (n=40) Conclusions: This pregnancy rate is much better than usual pregnancy rate expected after six embryos D3 or three embryos D5 transferred without pregnancy.

Outcomes of random-start letrozole protocol with PGT-A in women with breast cancer undergoing fertility preservation.

One hundred and seventeen who underwent SSCOS and 39 who underwent RSCOS for oocyte and/or embryo cryopreservation before breast cancer chemotherapy were retrospectively evaluated. Mean number of embryos and blastocyst euploidy rates were the main outcome measures. A majority of RSCOS cycles were initiated in the luteal phase (66.6% luteal vs. 33.3% follicular). While the total dose of gonadotropins was significantly higher in the RSCOS (3720.8 ± 1230.0 vs. 2345.1 ± 803.6IU; P < 0.001), the mean number of mature oocytes and embryos was similar to SSCOS. However, there was a trend for a higher number of mean embryos with luteal start RSCOS (6.9 ± 2.7 in late follicular start vs. 9.4 ± 4.2 in luteal start, P = 0.08). PGT-A was performed in 48% of the cases that underwent embryo cryopreservation in RSCOS (12 women, mean age = 35.3 ± 4.1; 87 blastocysts), revealing a euploidy rate of 36.2 ± 22.3% per patient. This rate was comparable to a 45% aneuploidy rate from similarly aged published data. Of the 7 RSCOS patients who returned for frozen embryo transfer, 5 delivered and one has an ongoing pregnancy, while in SSCOS, 18 out of 40 cycles resulted in live birth. Our data suggests that RSCOS fertility preservation cycle outcomes are similar to those with SSCOS and result in age-appropriate euploidy rates.

P-666 High embryo yield (and not oocyte yield) despite low ovarian stimulation dose correlates with cumulative livebirth rate in normal-high responders undergoing IVF/ICSI

Abstract Study question Does a good response i.e., high oocyte or embryo yield with low ovarian stimulation correlate with cumulative live birth outcome in IVF/ ICSI treatment? Summary answer High ovarian sensitivity to stimulation (embryos to gonadotropin dose ratio) positively predicts cumulative live birth in IVF, while oocytes to stimulation dose ratio does not. What is known already The conventional belief that the higher the number of eggs retrieved, the better for IVF outcome has been challenged. Oocyte yield fails to predict fresh-cycle live birth in presence of high ovarian response. Ovarian sensitivity index (OSI) which is an expression of inherent ovarian responsiveness to stimulation remained overlooked as a predictor of success in IVF. We explore whether attention should be on the oocyte or embryo yield as a function of ovarian stimulation (OSI) can predictor of positive outcomes in IVF. Study design, size, duration Retrospective analysis of 750 patients who underwent IVF/ ICSI treatment with ≤150 IU of gonadotropin in an antagonist protocol due to predicted normal or high response. This is a further analysis of a previously published study covering a period between 1 January 2016 to 31 Dec 2018. Participants/materials, setting, methods Multi-centre study within a group company using the same treatment protocols. Participants were normal or high responders of IVF/ ICSI, predicted poor responders (according to Bologna criteria) were excluded. All patients received ≤150 IU daily dose of gonadotropin (FSH) in an antagonist cycle. Cumulative LBR calculated. Univariate and multivariate (regression) analysis, adjusting for all significant variables were done to find the predictors of cumulative livebirth. Main results and the role of chance When the women who achieved a livebirth were compared with those who did not, there was no difference in the mean BMI (23.7 vs 23.6, p = 0.81), antral follicle counts (p = 0.33), Anti-mullerian hormone levels (p = 0,89), mean starting (149.5 vs 140.5 IU/ day, p = 0.16), and total gonadotropin doses (1360.7 vs 1342.8 IU, p = 0.62). Women’s age (33.5 vs 35.3 years, p &amp;lt; 0.0001) and the number of embryos transferred (p &amp;lt; 0.0001) had significantly correlation with LBRs. There was no difference in the mean number of oocyte (9.1 vs 8,6, p = 0.21) or oocyte yield per unit of gonadotropin (7.4 vs 6.7, p = 0.32).. The mean number of embryos (5.4 vs 4.1, p &amp;lt; 0.0001), transferable embryos (2.5 vs 1.9, p = 0.0006) and embryo yield per unit of gonadotropin (4.1 vs 3.2, p &amp;lt; 0.0001) were significantly higher in the livebirth group. After adjusting for woman’s age and the number of embryos transferred, the embryo yield (both total (p &amp;lt; 0.0001) and transferable (p = 0.01) embryos and embryo yield per gonadotropin unit (p = 0.0007) were predictors of cumulative livebirth whereas oocyte yield and oocyte number per unit of gonadotropin were not. The above findings remained the same when only IVF cycles excluding male infertility or only the cycles with a fixed 150 IU/d gonadotropin dose were analysed. Limitations, reasons for caution A moderate-sized study population. The poor responders have been excluded to minimise variations in the protocol and stimulation dose. Wider implications of the findings Our study is a proof of concept of ‘ovarian sensitivity index’ indicating the inherent ovarian responsiveness rather than the strength of stimulation determines the IVF/ICSI outcome. Embryo yield appeared to be a predictor of cumulative livebirth while oocyte yield is not, in normal and high responders of IVF. Trial registration number Not applicable

P-229 Prospective comparative study between two commercial heavy oils for culture using sibling donor oocytes collected

Abstract Study question To compare embryo development and clinical outcomes between two commercial heavy oils using sibling donor oocytes collected. Summary answer Our study suggests that both commercial heavy oils achieve similar embryo development and clinical outcomes rates. What is known already Current tendencies in IVF laboratories, such as extending the embryo culture uninterruptedly until day 6/7 or the use of dry time-lapse incubators, have enhanced the importance to use good quality oils supporting human embryo culture in vitro. The coating of the culture dishes with oil is highly important to maintain the ideal conditions that embryos need for an optimal development. Specifically, oil plays an essential role in maintaining a stable temperature, provides a barrier against external agents and contributes preventing the media evaporation, and thus, to the maintenance of an optimal pH and osmolality for the correct embryo development. Study design, size, duration This is a single-centre prospective study performed between February and November 2022 that included 180 donors and 213 recipients. Donors were randomized using a computer-generated randomization list. Each case was processed and cultured with a commercial single medium coated with a layer of the commercial heavy oil assigned, a mineral oil (A) or a paraffin oil (B). Participants/materials, setting, methods Oocytes were injected by ICSI and then cultured in 16-well dishes (EmbryoSlide+®, EmbryoScope+™,Vitrolife) prepared with each heavy oil (1700µl oil/dish). These were cultured in a time-lapse incubator (EmbryoScope+™,Vitrolife) at 37.29 ± 0.05 °C in an atmosphere of 6.5% CO2 and 5% O2. These parameters were controlled periodically (T+Button,BrightSentinel and G100,Geotech). Laboratory conditions, such as temperature, humidity and volatile organic compounds levels were monitored continuously (Octax Log&amp;Guard™,Vitrolife) during the study period, and pH was measured in a weekly basis. Main results and the role of chance A total of 2554 MII oocytes were injected by ICSI (oil A, n = 1304 and oil B, n = 1250). The proportion of fertilized oocytes was identical between the two oils (A:80.00% vs B:80.53%), as well as, abnormal fertilized oocyte rate (A:6.85% vs B:5.62%) and oocyte degeneration rate post-ICSI (A:6.14% vs B:6.00%). The mean number of embryos that reached the blastocyst stage and the proportion of blastocysts suitable for clinical use (transferred or cryopreserved) was almost the same independently of the oil used (A:69.84% vs B:67.15% and A:62.39% vs B:60.71%, respectively). Statistical data analysis was performed without referring to statistical significance (p &amp;gt; 0,05). 205 patients had an embryo transfer on day 5/6 with either fresh or cryopreserved blastocysts cultured coated with a layer of oil A (n = 101) or B (n = 104), with a mean number of 1.42 ± 0,55 and 1.33 ± 0,53 blastocysts transferred/patient in each group, respectively. No differences were found in terms of clinical pregnancy (A:69.30% vs B:67.31%) or implantation rates (A:62.37% vs B:61.53%) between both groups. Miscarriage rates were similar between group A (11.88%) and group B (12.62%). The pH average value during the study was 7.26 ± 0.06. The mean values of the room temperature, humidity and VOCs were stable at 21.7 ± 0.4 °C, 66.7 ± 6.9% and 0.098 ± 0.01ppm, respectively. Limitations, reasons for caution Although heavy oils are the most competent in keeping optimal culture conditions over time, there are several culture oils with different features available in IVF market. Thus, further studies should be performed comparing among them. Future research is also needed to compare peroxidation rates of our culture oils studied. Wider implications of the findings The present study suggests that both commercial heavy oils used in a continuous approach may provide similar in vitro fertilization rates regarding fertilization, blastocysts suitable for clinical use or clinical pregnancy. Heavy oil features, laboratory conditions and the culture environment should be properly validated independently on each IVF center. Trial registration number not applicable

Effect of breed and embryo sexing on the efficiency of a commercial OPU-ICSI program in mares

Breeders of Polo pony mares favour females over males as their sport performance is shown to be superior. This sex preference in breeding Polo horses is a challenge for stud veterinarians as sex sorted semen in horses is not available commercially due to technical difficulties. An alternative is the sexing of embryos. Ovum pick-up (OPU) and ICSI have become popular in show jumping (SJ) horses over the last decade, with the overall efficiency of the process improving steadily, with reported embryo per OPU session of > 1.5 embryos. The in vitro production of embryos not only allows a higher yield of blastocysts compared with in vivo derived embryos, but also their cryopreservation with similar pregnancy results similar to those achieved with fresh embryos. In addition, the possibility of sexing embryos before freezing would be an attractive option to the Polo breeding industry. Although there are several reports on the efficiency of an OPU-ICSI commercial program in SJ, information in Polo mares OPU-ICSI programs is scarce. The objective of this study was to describe the OPU-ICSI outcomes of a commercial program of Polo mares (n=77 cycles) compared to show jumping mares (n=73 cycles), including the likelihood of pregnancy after transferring sexed cryopreserved embryos. A general linear model of variance was created to test the effect of breed (Polo vs. SJ)on antral follicles available for aspiration, recovered oocytes, and number of embryos per OPU-ICSI session. Furthermore, the effect of embryo sexing prior to cryopreservation on pregnancy rate was determined by chi-square test. All OPUs were performed in the UK and oocytes were shipped to an external laboratory in Europe. Pre-implantation gender diagnosis was performed by extended embryo culture and sexing of extruded cells in 53 embryos of Polo mares. Polo mares had a lower (P<0.001) number of punctured follicles and recovered oocytes (14.1±0.6 and 8.2±0.5) compared to SJ mares (19.2±0.7 and 11.0±0.5), respectively. The mean number of embryos per OPU-ICSI session was similar (P>0.1) for SJ and Polo mares (1.38 and 1.41, respectively). Embryos derived from SJ mares (15/22, 68%) were more likely (P<0.05) to develop into a pregnancy than embryos from Polo mares (22/51, 43%). The ongoing pregnancy rate of sexed embryos (6/26, 23.1%) was lower (P=0.016) than that of unsexed embryos (14/25, 56%) from Polo mares, which explains the overall lower pregnancy outcome of Polo compared to SJ mares (unsexed embryos). In conclusion, Polo mares had a lower antral follicle pool than SJ mares, but their oocytes showed similar developmental potential to those of SJ mares. Embryo sexing decreased the chances of having a viable pregnancy after an OPU-ICSI session.

Gemellar Competition as a Key Component in Seed–Seedling Transition of Handroanthus chrysotrichus (Mart. ex A. DC.) Mattos (Bignoniaceae)

The occurrence of more than one embryo per seed (polyembryony) is common among angiosperms; however, there are gaps in the knowledge of its effects on the early stages of plant development. In this context, we study the effects of polyembryony and intraspecific variability in gemellar competition during the seed–seedling transition in Neotropical Handroanthus chrysotricus (Bignoniaceae). We used seeds from five cultivated trees in an urban environment inserted in a biodiversity hotspot (Cerrado). Embryo mass, seed germination, seedling emergence and seedling morphometry were evaluated. We did not find intraspecific variability in seed germination, seedling emergence or the mean number of embryos and seedlings per seed. On the other hand, intraspecific variability was observed during the transition from embryo to seedling. When only one seedling emerged from a seed, the seed–seedling transition was more asynchronous than when more seedlings emerged from one seed (with higher uncertainty and a longer time to emergence of the last seedling). The mass of embryos and seedlings decreased with the increase in the number of embryos in a seed, reinforcing the occurrence of gemellar competition. However, the total mass of embryos per seed was similar. The increase in seedlings per seed also decreased the morphometric measurements of each one. A positive morphometric aspect of the emergence of two seedlings per seed was that they had the highest total seedling mass, evidencing the positive Allee effect. Polyembryony had both positive and negative effects on seed germination and seedling morphology in the species, which helps to understand how this phenomenon acts on seed biology and plant establishment.

Comparison of aneuploidy rate in spontaneous abortion chorionic villus between D6 and D5 thawed-frozen blastocyst transfer

BackgroundTo compare the aneuploidy rate in spontaneous abortion chorionic villus (SA-CV) after D5 and D6 thawed-frozen blastocyst transfer(TBT).MethodsThis retrospective cohort study recruited 522 patients with early spontaneous abortion from March 2012 to January 2020 in the our center. The aneuploidy rate of SA-CV was compared according to the blastocyst development stage: D5 group (n = 398) and D6 group (n = 124).ResultsPatients’ characteristics, including age, body mass index, follicle-stimulating hormone, fertilization methods, type of infertility, infertility duration, and gestational age when abortion, did not differ between the two groups (all P > 0.05). Although the mean number of embryos was significantly higher in D6 than in the D5 group (P < 0.001), the mean number of high-quality embryos was similar (P = 0.773). In the D5 group, 46.5% of SA-CV showed aneuploidy, which was comparable to 41.1% in the D6 group (P = 0.296). After further grouping according to age (> 35 years or ≤ 35 years), the difference between the D5 and D6 groups remained not statistically significant (P = 0.247 and P = 0.690). Multivariate logistic analysis showed that women’s age was independently associated with the aneuploidy rate (OR = 0.891; 95% CI: [0.854–0.930]; P < 0.001). The rate of chromosomal aneuploidy was significantly higher in the age > 35 years group than in the age ≤ 35 years group (61.0% vs. 39.4%, P < 0.001). Other factors, including blastocyst formation speed, were not significant predictors of aneuploidy rate.ConclusionsThe rate of chromosomal aneuploidy in SB-CV after D6 TBT was comparable to that after D5 TBT. Chromosomal aneuploidy may not be a main factor contributing to the high prevalence early pregnancy loss at D6 group.

Presence of Endometrioma Does Not Impair Embryo Quality and Assisted Reproductive Technology (ART) Cycle Outcome in Diminished Ovarian Reserve (DOR) Patients.

This study aims to assess the impact of endometrioma on embryo quality and cycle outcome in patients who undergo assisted reproductive technology (ART) treatment due to diminished ovarian reserve (DOR). Retrospective case-control study was conducted in women ≤ 40years of age who underwent ART treatment caused by DOR, defined according to POSEIDON criteria, at a university-based infertility clinic between January 2015 and December 2020. Three groups of patients were selected: group A included patients with an idiopathic DOR, group B included patients with endometrioma(s) who underwent ovarian cystectomy, and group C included patients with endometrioma(s) without surgical treatment. A total of 351 women with DOR were included in the final analysis. Demographic characteristics, including age and AMH, were similar between the groups. Significant differences were observed among groups on mean number of MII oocytes retrieved (1.88 ± 1.59 vs. 2.84 ± 2.89 vs. 2.78 ± 2.41, respectively; p < 0.001) and mean number of embryos (1.04 ± 1.18 vs. 1.87 ± 2.01 vs. 1.66 ± 1.81, respectively; p < 0.001). However, the mean number of top-quality embryos, cycle cancellation, and live birth rates were similar between the groups. Clinical pregnancy (35 (26.5%) vs. 8 (18.2%) vs. 18 (42.9%), respectively; p = 0.038) and miscarriage rates (12 (9.1%) vs. 0 vs. 8 (19.0%), respectively; p = 0.009) were higher in endometrioma group without surgery. Women with DOR appear to have similar ART cycle outcomes regardless of the etiology, in terms of live birth rates. Infertility of endometrioma patients might be related to altered endometrium rather than to decreased oocyte quality. Cystectomy for endometrioma before IVF did not seem to affect the LBR.

P-729 The cumulative clinical pregnancy and live birth rates of IVF in women treated for genital tuberculosis are similar to those in other infertility etiologies

Abstract Study question Is cumulative clinical pregnancy rate (CPR) and live birth rate (LBR) in infertile patients with treated genital tuberculosis similar to other infertility etiologies undergoing IVF? Summary answer There is no significant difference in cumulative CPR and LBR in women adequately treated for genital tuberculosis undergoing IVF compared to other etiology of infertility. What is known already Female genital tuberculosis (FGTB) is an important cause of infertility in high tuberculosis prevalent countries. The uterine endometrial environment is an important cradle for embryo development. Once violated with infection, the damage to the endometrial blood flow and uterine cavity morphology is likely to be irreversible leading to infertility even after cure with anti-tuberculosis therapy (ATT). Adequate treatment of FGTB involves medical therapy with ATT and surgical treatment of hydrosalpinges and endometrial adhesions. The cumulative CPR and LBR are reported to be lower in the treated endometrial tuberculosis than in non-tuberculosis infertile patients undergoing IVF. Study design, size, duration This is a retrospective analysis of women who underwent IVF with self-oocytes over a period of 4 years from 2015 to 2018. All patients were divided into two groups as: Group-1: All women treated with ATT for FGTB before IVF with all presentations including women who underwent surgery for hydrosalpinges or intrauterine synechiae except with severe intrauterine adhesions and unfit for embryo transfer (ET) Group-2: Patients undergoing IVF other infertility etiologies. Participants/materials, setting, methods Women undergoing ovum pickup along with subsequent fresh and frozen embryo transfer attempts until attained a live birth or all the embryos were used. Exclusion criteria: Main results and the role of chance A total of 948 women underwent OPU during the study period. 171 were excluded from analysis for following reasons: Total of 775 women were included for analysis and results are summarized as: The mean number of embryos per OPU was 7.04 in Group-1 while 6.97 in Group-2. A total of 195 ET procedures were done in Group-1 while 1035 in Group-2. The mean number of ET procedures required for one clinical pregnancy in Group-1 was 3.15 and 2.81 in Group-2.The mean number of ET procedures required for one live birth in Group-1 was 3.42 and 3.02 in Group-2. CPR per ET procedure in Group-I was 31.79% while 35.55% in Group-2. All these differences were statistically insignificant. Limitations, reasons for caution The FGTB has a wide spectrum of presentation from a very early sub-clinical disease to an advanced stage with pelvic organ damage. The prognosis of IVF varies with the extent of disease. This study analyzed overall result while sub categories of FGTB were not analyzed. Wider implications of the findings Infertility after ATT treated FGTB has good IVF outcome. IVF outcome in these patients need to be studied based upon the extent of damage. We recommend undertaking an early IVF with good hope of live birth in ATT treated FGTB. Trial registration number Not applicable

O-068 The BISTIM study: first RCT comparing dual ovarian stimulation on the same cycle (duostim) vs 2 conventional ovarian stimulations in poor ovarian responders undergoing IVF

Abstract Study question Is the number of cumulated oocytes with dual ovarian stimulation on the same cycle (duostim) higher compared to 2 consecutive antagonist cycles in poor responders? Summary answer Considering the number of total and mature oocytes collected in poor ovarian responders, there is no benefit of duostim vs two consecutive antagonist cycles. What is known already Several waves of follicular development exist on the same cycle. Recent studies have shown the ability to obtain oocytes with equivalent quality in the luteal phase, even after a previous ovarian stimulation in the follicular phase. During stimulation, smaller follicles are recruited and sensitized, which may increase the selection of follicles available on the second stimulation. In poor ovarian responders (POR) this potentialization may have a great interest, as 2 stimulations on the same cycle could give a higher number of oocytes compared to two conventional stimulations. However, these preliminary data need to be confirmed with a randomized controlled trial. Study design, size, duration This is a multicenter, open-labeled randomized control trial (2018, september – 2021, march). The primary objective was to demonstrate that two ovarian stimulations within the same cycle (first in the follicular phase, followed by a second in the luteal phase) lead to the retrevial of 1.5 more oocytes than the cumulative number of oocytes from two consecutive conventional stimulation, in POR women. According to this hypothesis, 44 patients were needed in each group. Participants/materials, setting, methods 88 POR women, defined with Bologna criteria (AFC≤5 and/or AMH≤1.2ng/ml and ≤3 oocytes if previous IVF) were randomized, 44 in duostim group (D) and 44 in conventional group (C). FertistartKit® 300IU/day with antagonist protocol was used except in luteal phase stimulation of group D. In group D, oocytes were pooled and inseminated after the second retrieval, with freeze all embryos. Fresh transfer was performed in group C. The analysis is presented in intention to treat. Main results and the role of chance There was no difference between the groups regarding demographics, ovarian reserve markers (AFC, AMH) and stimulation parameters. The mean number of cumulated oocytes retrieved with 2 ovarian stimulation was not statistically different in group D and C, respectively 5.0+/-3.4 and 4.6+/-3.4 (p = 0.56). The mean number of cumulated mature oocytes was not statistically different, 3.7+/-3.3 in group D vs 3.1+/-3.0 in group C (p = 0.38). The mean number of embryos was significantly lower in the group D, 0.8+/-1.3 vs group C 1.6+/-1.3 (p &amp;lt; 0.01). There was no statistical difference of the mean number of oocytes retrieved per cycle in cycle 1 vs cycle 2 in both group D and C. The delay, between the first and the second day 1 of stimulation was statistically different in group D 14.4 days (10-19) vs group C 90.6 (28-232). The ongoing pregnancy rate in group D 17.9% (7/39) was not statistically different with group C 29.3% (12/41), (p = 0.23). And the mean time to ongoing pregnancy tends to be longer in group D (144 days) vs group C (115 days) but was not statistically different (p = 0.21). Limitations, reasons for caution The RCT was impacted by Covid pandemia and stop of IVF activities for 10 weeks. Delays were recalculated to exclude this period, however one women in group D cannot have the luteal stimulation. We also faced unexpected good ovarian responses and pregnancies after the first oocyte pick-up in group C. Wider implications of the findings In routine practice, the benefit of duostim in patients with POR is not confirmed. Firstly, because there is no potentialization on the number of oocyte retrieved in luteal phase after follicular phase stimulation. Secondly, because the freeze all strategy avoids a pregnancy with fresh embryo transfer after the first cycle. Trial registration number 2017-A00498-45

Morphokinetic changes in vitrified and non-vitrified in vitro-derived ovine embryos

The present experiment employed time-lapse (TL) imaging to assess the effects of vitrification on the development of ovine blastocysts and to see if the timing of blastocyst formation and expansion was correlated with the numbers of embryo- and trophoblasts determined through differential staining of the expanded blastocysts. Ovaries were obtained after slaughter from cycling (October–March) Polish Longwool ewes aged 1–3 years and cumulus-oocyte complexes were collected by ovarian scarification. In vitro maturation was performed in TCM 199 medium supplemented with Earle's Salt, 10% of FBS, and 5 μg/mL of LH/FSH at 38 °C for 24 h. After maturation, the oocytes were incubated with thawed ram semen (IVF) for 19 h and all presumptive zygotes were transferred to a 16-well dish containing Cult medium for monitoring with the Primo Vision TL system. A portion of ovine embryos were vitrified (Cryotop system) at the early cavitation stage and TL observations of warmed (n = 30) and non-vitrified (n = 32) blastocysts continued until the attainment of the expanded blastocyst stage, at which point they were differentially stained with bisbenzimide and propidium iodide for microscopic enumeration of embryoblasts (inner cell mass blastomeres) and trophoblast-cells. There were no statistically significant differences in the timing of blastulation (tB) and formation of expanded blastocysts (tEB) between vitrified and non-vitrified ovine embryos, but non-vitrified blastocysts exceeded (P < 0.05) their vitrified and warmed counterparts in the mean number of embryo- and trophoblasts. In addition, the number of trophoblasts was negatively and moderately correlated with tB and tEB, for both vitrified and non-vitrified embryos. It can be concluded that even though vitrification of ovine embryos is associated with a significant reduction in the number of blastomeres, the rate of blastocyst development remains closely linked to the numbers of trophoblastic cells.

Impact of comprehensive state insurance mandates on in vitro fertilization utilization, embryo transfer practices, and outcomes in the United States

Previous studies have demonstrated that state mandated coverage of invitro fertilization may be associated with increased utilization, fewer embryos per transfer, and lower multiple birth rates, but also lower overall live birth rates. Given new legislation and the delay between enactment and effect, a revisit of this analysis is warranted. This study aimed to characterize the current impact of comprehensive state invitro fertilization insurance mandates on invitro fertilization utilization, live birth rates, multiple birth rates, and embryo transfer practices. We conducted a retrospective cohort study of invitro fertilization cycles reported by the 2018 Centers for Disease Control and Prevention Assisted Reproductive Technology Fertility Clinic Success Rates Report in the United States. Invitro fertilization cycles were stratified according to state mandate as follows: comprehensive (providing coverage for invitro fertilization with minimal restrictions) and noncomprehensive. The United States census estimates for 2018 were used to calculate the number of reproductive-aged women in each state. Outcomes of interest (stratified by state mandate status) included utilization rate of invitro fertilization per 1000 women aged 25 to 44 years, live birth rate, multiple birth rate, number of embryo transfer procedures (overall and subdivided by fresh vs frozen cycles), and percentage of transfers performed with frozen embryos. Additional subanalyzes were performed with stratification of outcomes by patient age group. In 2018, 134,997 invitro fertilization cycles from 456 clinics were reported. Six states had comprehensive mandates; 32,029 and 102,968 cycles were performed in states with and without comprehensive invitro fertilization mandates, respectively. Invitro fertilization utilization in states with comprehensive mandates was 132% higher than in noncomprehensive states after age adjustment; increased utilization was observed regardless of age stratification. Live birth rate per cycle was significantly higher in states with comprehensive mandates (35.4% vs 33.4%; P<.001), especially among older age groups. Multiple birth rate as a percentage of all births was significantly lower in states with comprehensive mandates (10.2% vs 13.8%; P<.001), especially among younger patients. Mean number of embryos per transfer was significantly lower in states with comprehensive mandates (1.30 vs 1.36; P<.001). Significantly fewer frozen transfers were performed as a percentage of all embryo transfers in states with comprehensive mandates (66.1% vs 76.3%; P<.001). Among fresh embryo transfers, significantly fewer embryos were transferred in comprehensive states among all patients (1.55 vs 1.67; P<.001). Comprehensive state mandated insurance coverage for invitro fertilization services is associated with greater utilization of these services, fewer embryos per transfer, fewer frozen embryo transfers, lower multiple birth rates, and higher live birth rates. These findings have important public health implications for reproductive-aged individuals in the United States and present notable opportunities for research on access to fertility care.

Direct somatic embryogenesis and flow cytometric assessment of ploidy stability in regenerants of Caladium × hortulanum ‘Fancy’

Caladium × hortulanum 'Fancy' is an important ornamental plant grown in pots and landscapes and known for its colorful leaves often used for interior decorations. In this work, we present a method of in vitro regeneration from three explants source through direct somatic embryogenesis (DSE) wherein the regenerated plants were screened for ploidy changes through flow cytometry analysis. Tuber, leaf and petiole explants were cultured on MS basal medium supplemented with 1-napthalene acetic acid (NAA), 6-benzyl amino purine (BAP) and N-phenyl-N'-1, 2,3-thiadiazol-5-ylurea (TDZ) concentrations. Tuber explants induced highest direct somatic embryos on NAA (1 mg L- 1) + BAP (0.5 mg L- 1) with 55.6 mean number of embryos per explant while as leaf and petiole explants amended with 1 mg L- 1 TDZ developed 18.7 and 12.27 mean number of embryos per explant respectively. The highest embryo conversion frequency was achieved on BAP (2 mg L- 1) + NAA (0.2 mg L- 1) with 44.2, 18.7 and 7.5 mean number of plantlets produced per tuber, leaf and petiole explant respectively after 4 weeks of culture. Plantlets were later rooted and maximum number of roots (6.33) per shoot was achieved on 2 mg L- 1 indolebutyric acid amended medium. Description of the process of DSE is presented through the histological and SEM evidences. The 2C DNA content of field grown plants and the DSE regenerants evaluated under flow cytometric analysis were 8.06 pg and 8.28 pg respectively showing no ploidy changes. Hence, a successful protocol of inducing direct somatic embryos from three explant types with efficient embryo conversion frequency was obtained with regenerants showing similar DNA ploidy as that of their parent plants.

IMPACT OF STATE IVF INSURANCE MANDATES ON EMBRYO TRANSFER PRACTICES IN THE UNITED STATES

To characterize the impact of state IVF insurance mandates on embryo transfer practices in terms of mean number of embryos per transfer and utilization of fresh versus frozen embryo transfers. Data from the 2018 CDC ART Fertility Clinic Success Rates Report and the National ART Surveillance System (NASS) were used to calculate the number of embryos transferred per procedure and number of fresh and frozen embryo transfers. Data were stratified into two groups, “Comprehensive” and “Non-comprehensive,” based on the degree of IVF coverage mandated by state laws. Two-sample t-test and chi-square test were used to compare mean number of embryos transferred per procedure and frozen transfers as a share of all transfers, respectively, according to mandate status. In 2018, 456 clinics reported data to the CDC. Comprehensive IVF mandates were identified in 6 states. Mean number of embryos per transfer was significantly lower in states with comprehensive IVF mandates compared to states without these mandates among all transfer types. On age-group sub-analysis, this difference remained significant among transfers performed in younger age groups. Significantly more embryos were transferred in fresh than frozen embryo transfers. Significantly fewer frozen transfers as a share of all embryo transfers were performed in states with comprehensive IVF mandates.Tabled 1State Mandate to Cover IVFEmbryos per Transfer, AllOverallAge <35Ages 35-37Ages 38-40Ages 41-42Age >42Comprehensive1.301.181.241.401.681.88Non-comprehensive1.361.291.311.421.591.84P-value<0.001<0.001<0.0010.02<0.0010.3Embryos per Transfer, FreshComprehensive1.551.251.411.702.252.50Non-comprehensive1.671.421.601.892.182.30P-value<0.001<0.001<0.001<0.0010.090.04Embryos per Transfer, FrozenComprehensive1.181.151.161.191.311.44Non-comprehensive1.271.251.231.271.331.57P-value<0.001<0.001<0.001<0.0010.3<0.001% Frozen Transfers PerformedComprehensive66%67%69%67%59%54%Non-comprehensive76%76%79%77%71%66%P-value<0.001<0.001<0.001<0.001<0.001<0.001 Open table in a new tab Comprehensive IVF insurance mandates were associated with a significantly lower mean number of embryos transferred per procedure. More embryos were transferred in fresh compared to frozen embryo transfers. Frozen embryo transfers accounted for smaller share of embryo transfers in states with comprehensive IVF mandates.

FAILED THAW CYCLES (FTHC) AS A KEY PERFORMANCE INDICATOR (KPI): EVALUATING THE PROGRESS OF FROZEN EMBRYO TRANSFERS (FET) IN USA

Embryo cryosurvival rate is a recommended KPI to be benchmarked. Lower success rates in FET cycles could be related to ET decisions and freezing procedures, including minimal standards to cryopreserve. We evaluated FThC cycles and related FET outcomes across SART age groups and infertility diagnosis over 6 years. Descriptive review of the SART database included 262,818 FET thawing cycles, 2014-2019 (1st transfer ≥12 months after retrieval, 2nd or later transfers only, limited by database output). Outcomes were: percentage of thaw cycles with no embryos suitable for transfer (FThC), mean number of embryos transferred, and live birth rate (LBR). Regression models adjusting for age and diagnosis estimated the changes in these outcomes over time. Mean increase in thaw cycles was 15.2% per year (32,381 to 57,019, 2014-2019). FThC decreased for all age groups, -0.28% per year (p<0.0001), independent of diagnosis; FThC 2014 to 2019: <35: 1.1% to 0.4%; 35-37: 1.5% to 0.6%; 38-40: 1.5% to 0.8%; 41-42: 2.4% to 1.0%; >42: 3.3% to 2.4% - highest improvement of 61% for ages 41-42. Mean number of embryos transferred decreased from 1.62 to 1.22 (p<0.0001). LBR increased across all age groups by +2.17% per year (p<0.0001). Age ≥41 had over 10% improvement and younger groups had over 5% improvement. LBR also increased for all diagnoses combined by +1.76% per year (p<0.0001), and for individual diagnoses by 1.14–3.53% (all p<0.01), but didn’t reach significance for uterine factors (Table). FThC significantly decreased across all ages and diagnosis, even as FET cycles increased. This likely reflects the evolution of freezing criteria, procedures and clinical decisions positively impacting LBR. Such improvements in FThC and LBR along with fewer embryos transferred may also reflect the impact of PGT and freeze-all. This additional analysis is underway.

P–448 Clinical outcome of social oocyte cryopreservation at advanced age

Abstract Study question What are the success rates of social oocyte cryopreservation (SOC) at advanced age? Summary answer In this study, one in four women who underwent SOC above age 35 had a delivery. What is known already While SOC is gaining popularity, reports on delivery rates are limited due to low utilization rates. Study design, size, duration Retrospective data collection of all woman who underwent SOC between 2011–2018, and presented for treatment using cryopreserved oocytes until January 2021. Participants/materials, setting, methods: Review of patient records (including both IVF and antenatal/postnatal) and laboratory data in a university affiliated hospital-based IVF unit. Main results and the role of chance: A total of 448 women underwent SOC during 2011–2018. 50 (11.2%) women returned to use these oocytes until the end of January 2021. Women who returned to use their oocytes underwent cryopreservation at mean age of 38.2±2.2. 46 (92%) of participants were above 35 at time of cryopreservation. Number of oocytes cryopreserved was 11.3±9.7. Mean time from cryopreservation to thawing was 5.5±1.8years (range 1–9 years). and age at thawing was 43.4±2.1 (range 40–49). Nearly half of patients initially attempted to conceive before using their cryopreserved oocytes, mostly by ART using fresh oocytes. Mean number of oocytes thawed and oocytes survived per women was 9.7±6.2 and 6.1±4.9 respectively (post thawing survival rate 65.4±35%). Mean number of embryos transferred, at one or more attempts was 2.6±2.1 per women. Eleven women gave birth or had an ongoing pregnancy &amp;gt; 20 weeks at time of analysis. All deliveries resulted from cryopreservation at age 36 and older (delivery rate 23.9% per women). Limitations, reasons for caution: We report our initial experience of women who underwent SOC at a single center. Most women who returned to use their oocytes had undergone SOC at advanced age, therefore not necessarily reflecting outcome for younger patients attempting to preserve fertility using this technology. Wider implications of the findings: Considering modest success rates of SOC in our cohort, women considering SOC are advised to do so at an earlier age. Trial registration number Not applicable

P–667 Dydrogesterone supplementation in cycles triggered with lone GnRH agonist for final oocyte maturation resulted in an acceptable pregnancy rate

Abstract Study question Can luteal oral Dydrogesterone (Duphaston) supplementation in an antagonist cycle after a lone GnRH agonist trigger rescue the luteal phase, allowing the possibility to peruse with fresh embryo transfer? Summary answer Functionality of the luteal phase in an antagonist cycle after a lone GnRH agonist trigger can be restored by adding Duphaston to conventional luteal support. What is known already Ovarian hyperstimulation syndrome (OHSS) is dramatically reduced when using antagonist cycle with lone GnRH agonist trigger before ovum pick up. This trigger induces short luteinizing hormone (LH) and follicle-stimulating hormone (FSH) peaks, associated with reduced progesterone and estrogen levels during the luteal phase. They cause an inadequate luteal phase and a significantly reduced implantation rate leading to a freeze all practice in those cycles. Study design, size, duration A retrospective cohort study. The study group (n = 123) included women that underwent in vitro fertilization cycles from January 2017 to May 2020. Patients received a GnRH-antagonist with a lone GnRH-agonist trigger due to imminent OSHH. The control group (n = 374) included patients under 35 years old that, during the same time period, underwent a standard antagonist protocol with a dual trigger of a GnRH-agonist and hCG. Participants/materials, setting, methods Study patients were given Dydrogesterone (Duphaston) in addition to micronized progesterone vaginal pills (Utrogestan) for luteal support (Duphaston group). Controls were treated conventionally with Utrogestan for luteal phase support (hCG group). The outcomes measured were pregnancy rate and OHSS events. Main results and the role of chance Our study was the first to evaluate the addition of Duphaston to standard luteal phase support in an antagonist cycle triggered by a lone GnRH agonist before a fresh embryo transfer. The mean number of oocytes retrieved and estradiol plasma levels were significantly higher in the Duphaston group than in the hCG group (16.9 ±7.7 vs. 10.8 ± 5.3 and 11658 ± 5280 pmol/L vs. 6048 ± 3059 pmol/L, respectively). The fertilization rate was comparable between the two groups. The mean number of embryos transferred and the clinical pregnancy rate were also comparable between groups (1.5 ± 0.6 vs 1.5 ± 0.5 and 46.3% vs 40.9%, respectively). No OHSS event was reported in either group. Limitations, reasons for caution This retrospective study may carry an inherent selection and information bias, derived from medical record coding. An additional limitation was the choice of physician for the lone GnRH trigger, which may have introduced a selection bias and another potential caveat was the relatively small sample size of our study groups. Wider implications of the findings: The addition of Duphaston to conventional luteal support could effectively salvage the luteal phase without increasing the risk for OHSS. This enables, to peruse in those cycle, with fresh embryo transfer, avoiding the need to freeze all the embryos and postponed embryo transfer. Leading to lower psychological burden and costs. Trial registration number 0632–20-HMO

Effects of temperature pretreatments and carbohydrate type on androgenesis induction in anther culture of Capparis spinosa L.

Caper (Capparis spinosa L.) is a rich source of rutin, plays an essential role in human health. In the present study, the effects of cold (25°C as control, 4°C, and 7°C for 2, 4, and 7 days), heat (25°C as control, 30°C for 14 days, 32°C for 2 and 4 days, and 35°C for 8 hours), and carbohydrate treatments on the androgenesis efficiency were studied in the anther culture of caper. Also, the effects of maltose and sucrose at the concentrations of 30 and 60 g L-1 in combination with two temperature treatments (1- 30°C for 14 days and 2- 7°C for 7 days + azacytidine and 2,4-D pretreatments) on the androgenesis induction was evaluated. The temperature and carbohydrate treatments showed statistically significant differences (p ≤0.01) in terms of callus and embryo formation. The 7°C for 2, 4, and 7 days produced the highest percentage (at the third week: 80, 78.34, and 76.67%, respectively) and callogenesis speed (7.85, 7.75, and 7.60 calli week-1, respectively) and the 7°C for 7 days produced the highest embryo production (0.57 embryo anther-1). The 30°C for 14 days treatment showed the highest percentage (at the third week: 100%) and callogenesis speed (9.44 calli week-1). While the 32°C for 2 and 3 days and also 30°C for 14 days produced the highest number of embryos per anther (0.22, 0.20, and 0.18 embryo, respectively). The use of 30 g L-1 maltose in combination with the 30°C for 14 days produced the highest percentage (at the third week: 91.66%) and callogenesis speed (8.94 calli week-1), while the 30 g L-1 maltose in combination with the 7°C for 7 days + azacytidine and 2,4-D pretreatments produced the highest mean embryo number per anther (0.55 embryo). The results of this research are of great importance for the use in the caper breeding programs.

Preimplantation genetic testing for aneuploidies (abnormal number of chromosomes) in in vitro fertilisation.

Preimplantation genetic testing for aneuploidies (abnormal number of chromosomes) in in vitro fertilisation.