Top-quality Embryos Research Articles

Does daily co administration of gonadotropins and letrozole during the ovarian stimulation improve IVF outcome for poor and sub optimal responders?

BackgroundCo-administration of letrozole during the first 5 days of ovarian stimulation was suggested to improve IVF outcomes in poor responders. We aimed to determine whether poor/sub-optimal responders might benefit from Letrozole co-treatment throughout the entire stimulation course.MethodsWe retrospectively reviewed the medical files of women who demonstrated poor (oocyte yield ≤3) and sub-optimal (4 ≤ oocyte yield ≤9) ovarian response during conventional multiple-dose antagonist stimulation protocols and were co-treated in a subsequent cycle with 5 mg Letrozole from the first day of stimulation until trigger day. A self-paired comparison between gonadotropins-only and gonadotropins-letrozole cycles was performed.ResultsTwenty-four patients were included. Mean patients’ age was 39.83 ± 4.60 and mean day-3-FSH was 12.77 ± 4.49 IU/m. Duration of stimulation and total gonadotropins dose were comparable between the two cycle groups. Peak estradiol levels were significantly lower in gonadotropins-letrozole cycles (2786.74 ± 2118.53 vs 1200.13 ± 535.98, p < 0.05). Number of retrieved oocytes (3.29 ± 2.15 vs 6.46 ± 3.20, p < 0.05), MII-oocytes (2.47 ± 1.65 vs 5.59 ± 3.20, p < 0.05), 2PN-embryos (1.78 ± 1.50, 4.04 ± 2.74, p < 0.05) and top-quality embryos (0.91 ± 0.97 vs. 2.35 ± 1.66, p < 0.05) were significantly higher in the gonadotropins-letrozole cycles. Clinical pregnancy rate in gonadotropins-letrozole cycles was 31.5%.ConclusionLetrozole co-treatment during the entire stimulation course improves ovarian response and IVF outcomes in poor/sub-optimal responders.

Effect of serum 25-hydroxyvitamin D levels on sperm quality and assisted reproductive technology outcomes for men of infertile Chinese couples.

Vitamin D insufficiency/deficiency is highly prevalent in China, but its effect on sperm quality and outcomes of assisted reproductive technology (ART) in men from infertile couples is not clear. This study aimed to explore the association of serum 25-hydroxyvitamin D (25OHD) levels and semen parameters, as well as ART outcomes for men from infertile couples. This study recruited 1308 men from infertile couples who attended the Reproductive Medicine Center of Peking University Third Hospital for first time from February to August 2019 (NCT03804294). Reproductive hormones levels, semen parameters, and clinical outcomes of ART were compared between vitamin D-deficient group (serum 25OHD≤10ng/mL), insufficient group (serum 25OHD 10-20ng/mL), and sufficient group (serum 25OHD≥20ng/mL). We found 27.7% of participants were vitamin D sufficient, suggesting high prevalence of vitamin D insufficiency/deficiency in China. Reproductive hormone levels were similar between the three groups. The total number of morphologically normal spermatozoa was significantly different between the three groups in all participants, impaired semen quality subgroup, and normozoospermia subgroup (P<.05) with the adjustment of confounders (age, BMI, alcohol consumption, cigarette smoking, and physical exercise). The percentage of normal sperm morphology significantly increased with elevated serum 25OHD levels in all participants and the participants with impaired semen quality (P<.05), and the total sperm count showed significant difference in participants with impaired semen quality (P=.026). For routine in vitro fertilization and intracytoplasmic sperm injection cycles, there were no differences in fertilization rate, top-quality embryo rate, biochemical pregnancy rate, and clinical pregnancy rate between serum 25OHD<20ng/mL and serum 25OHD≥20ng/mL groups. Males of infertile couples with higher serum 25OHD levels exhibit better results in sperm morphology, and serum 25OHD levels may contribute to total sperm number in participants with impaired semen quality, but show no effects on the ART outcomes.

Do Follicles of Obese Patients Yield Competent Oocytes/Embryos?

Aim: This study evaluated the competency of oocytes/embryos derived from follicles >15 mm in diameter from obese patients, compared with nonobese patients. Patients and Methods: A cohort study was conducted in a single tertiary medical center between July 2018 and May 2019. Before ultrasound-guided follicular aspiration, follicles were measured and those with maximal dimensional size >15 mm were tracked. Microscopic examination of the follicular aspirates was performed by an embryologist. Each follicle aspirated was evaluated for oocyte maturation, oocyte fertilization, and embryo quality. Results: 457 follicles were measured: 380 (83.2%) in nonobese and 77 (16.8%) in obese patients. No in-between group differences were observed in the causes of infertility, patients’ demographics, or ovarian stimulation characteristics. Oocytes were achieved during aspiration from 277 (72.8%) and 54 (70.0%) of the nonobese and obese groups, respectively (p = 0.67). No in-between group differences were observed in fertilization (2PN/oocyte), top quality embryo (TQE) per zygote (2PN), and TQE per follicle. Conclusion: Oocyte recovery rate from follicles >15 mm is unrelated to patients’ BMI. Moreover, the oocytes recovered from obese patients are competent yielding comparable zygote and TQE per follicle/oocyte, compared with nonobese patients. Further investigation is required to strengthen this finding.

Associations of post-warming embryo or blastocyst development with clinical pregnancy in vitrified embryo or blastocyst transfer cycles.

Objective To investigate whether the degree of post-warming embryo or blastocyst development is associated with clinical pregnancy in vitrified embryo or blastocyst transfer cycles.Methods Ninety-six vitrified cleavage-stage embryos and 58 vitrified blastocyst transfer cycles were selected. All transfer cycles were performed from February 2011 to March 2019, and all vitrified embryos or blastocysts were warmed from 4 PM to 6 PM and then transferred the next morning from 9 AM to 10 AM. The scores of the cleavage-stage embryos and blastocysts were assessed at warming and at transfer using the modified Steer method and the Gardner method, respectively. The mean embryo or blastocyst score, score of the single top-quality embryo or blastocyst, and the difference in the score between warming and transfer were compared between nonpregnant and pregnant women.Results In the cleavage-stage embryo transfer cycles, both the top-quality embryo score at transfer and the difference in the score between warming and transfer were significantly associated with clinical pregnancy. A top-quality embryo score at transfer of ≥60.0 (area under the curve [AUC], 0.673; 95% confidence interval [CI], 0.531–0.815) and a difference in the score between warming and transfer of ≥23.0 (AUC, 0.675; 95% CI, 0.514–0.835) were significant predictors of clinical pregnancy. In blastocyst transfer cycles, the top-quality blastocyst score at transfer was the only significant factor associated with clinical pregnancy. A top-quality blastocyst score at transfer of ≥38.3 was a significant predictor of clinical pregnancy (AUC, 0.666; 95% CI, 0.525–0.807).Conclusion The top-quality embryo score at transfer and the degree of post-warming embryo development were associated with clinical pregnancy in vitrified cleavage-stage embryo transfer cycles. In vitrified blastocyst transfer cycles, the top-quality blastocyst score at transfer was the only significant factor affecting clinical pregnancy.

Stop GnRH-Agonist Combined With Multiple-Dose GnRH-Antagonist Protocol for Patients With "Genuine" Poor Response Undergoing Controlled Ovarian Hyperstimulation for IVF.

Objective: To examine whether the Stop GnRH-agonist combined with multiple-dose GnRH-antagonist protocol may improve conventional IVF/intracytoplasmic sperm injection (ICSI) cycle in poor ovarian response (POR) patients.Design: Cohort historical, proof of concept study.Setting: Tertiary, University affiliated Medical Center.Patient(s): Thirty POR patients, defined according to the Bologna criteria, who underwent a subsequent Stop GnRH-agonist combined with multiple-dose GnRH-antagonist controlled ovarian hyperstimulation (COH) protocol, within 3 months of the previous failed conventional IVF/ICSI cycle, were included. For the purposes of this study, we eliminated a bias in this selection by including only “genuine” poor responder patients, defined as those who yielded up to 3 oocytes following COH with a minimal gonadotropin daily dose of 300 IU.Main Outcome Measure(s): Number of oocytes retrieved, number of top-quality embryos, COH variables.Result(s): The Stop GnRH-agonist combined with multiple-dose GnRH-antagonist COH protocol revealed significantly higher numbers of follicles >13 mm on the day of hCG administration, higher numbers of oocytes retrieved, and top-quality embryos (TQE) with an acceptable clinical pregnancy rate (16.6%). Moreover, as expected, patients undergoing the Stop GnRH-agonist combined with multiple-dose GnRH-antagonist COH protocol required significantly higher doses and a longer duration of gonadotropins stimulation.Conclusion(s): The combined Stop GnRH-ag/GnRH-ant COH protocol is a valuable tool in the armamentarium for treating “genuine” poor ovarian responders. Further, large prospective studies are needed to elucidate its role in POR and to characterize the appropriate patients subgroup (before initiating ovarian stimulation) that may benefit from the combined Stop GnRH-ag/GnRH-ant COH protocol.

Reduced ovarian response to controlled ovarian stimulation is associated with increased oxidative stress in the follicular environment

Serum estradiol (E2) level is routinely used to monitor the ovarian response during controlled ovarian hyperstimulation (COH) and the concentration of serum E2 may influence the oocyte quality and pregnancy outcome. However, the knowledge on the association between COH induced serum E2 level, oocyte quality and embryo development is limited. Therefore we investigated the association between serum E2 level, oxidative stress in the follicular fluid and granulosa cells (GCs) response to elucidate the association between E2 level and embryological outcome. In this study, patients (n = 30) undergoing ART were categorized as ‘normal responders’ (NR, n = 10), ‘poor responders’ (PR, n = 10) and hyper responders (HR, n = 10). The follicular fluid malondialdehyde (MDA) level was determined. The total RNA extracted from GCs was subjected to analyse the relative abundance of transcripts of stress response genes (P53, caspase 3,8-oxoguanine DNA glycosylase, OGG1 and heat shock protein 70; HSP70) and embryological outcome was noted. Follicular fluid MDA level was significantly higher in PR (p < 0.01) compared NR and HR whereas number of top-quality embryos were significantly lower in PR and HR compared to NR (p < 0.01). The relative expression of P53, HSP70, and OGG1 in GCs was significantly elevated in PR (p < 0.05−0.01). An inverse relationship was established between serum E2 level vs follicular MDA level (r = −0.45; p < 0.01) and follicular MDA level vs. number of top-quality embryos (r = −0.45; p < 0.01). Hence, patients with low serum E2 had elevated oxidative stress in their follicular environment and poor quality embryos implicating the risk of oxidative stress in patients with poor ovarian response.

GnRH agonis short protocol versus GnRH antagonist flexible protocols in poor responder undergoing in vitro fertilization treatment.

Objective: To compare the short agonist and flexible antagonist protocols in the management of poor responders to standerd long down regulation protocol. Design: Randomized prospective comparative study. Setting: private IVF center(Engab). Patient(s): One hundred poor-responder (IVF)(ICSI), who responded poorly to the standerd long down- regulation protocol in their first treatment cycle. Intervention(s): Peak serum E2 was assayed on the day of hCG administration. Main Outcome Measure(s): primary outcome: number of HMG ampoules used, peak E2 at the triggering day, number of oocytes retrieved, number of metaphase II oocytes, fertilization rate, embryo quality and number of embryos transferred.cycle cancellation. secondary outcome: pregnancy rate , miscarriage rate . Result(s): GnRH-antagonist protocol shows increased – non significant- pregnancy rate and decreased - non significant - miscarriage rate than agonist short protocol, however agonist short protocol appears to be more effective than the GnRH-antagonist protocol in terms of number of FSH ampules, peak E2 level, fertilization rate, and top-quality embryos transferred in poor-responder. Conclusion(s): The probability of pregnancy might be better with the use of the antagonist protocol, also flexible antagonist protocol is a simpler and more patient friendly method of ovarian stimulation compared with the short agonist protocol.

Obstetric and perinatal risks in 4601 singletons and 884 twins conceived after fresh blastocyst transfers: a Nordic study from the CoNARTaS group.

Are obstetric and perinatal outcomes in pregnancies after fresh blastocyst transfer (BT) comparable with those born after fresh cleavage stage transfer (CT) and spontaneous conception (SC)? Fresh BT is associated with a higher risk of placental and perinatal complications. BT optimizes the selection of top-quality embryos and increases pregnancy and live birth rates per transfer compared to CT. However, concerns have been raised as extended culture duration may increase obstetric complications and impair perinatal outcomes. Previous studies have shown a higher risk of preterm birth (PTB) among infants born after BT compared with CT. Pregnancies after BT are also prone to a higher risk of same-sex twins after single embryo transfer (SET). A retrospective register-based cohort study used data from Denmark, Norway and Sweden including three cohorts: 56 557 singletons and 16 315 twins born after fresh IVF/ICSI cycles and 2 808 323 SC singletons in Denmark (birth years 1997-2014), Norway (2010-2015) and Sweden (2002-2015). Of the fresh IVF/ICSI singletons, 4601 were born after BT and 51 956 after CT. The twin cohort consisted of 884 fresh IVF/ICSI children born after BT and 15 431 fresh IVF/ICSI children born after CT. Data were obtained from a large Nordic cohort of children born after ART and SC initiated by the Committee of Nordic ART and Safety (CoNARTaS). The CoNARTaS cohort was established by cross-linking National ART-, Medical Birth-, and National Patients Registers using the unique personal identification number, allocated to every citizen in the Nordic countries. Obstetric and perinatal outcomes after BT, CT and SC were compared using logistic regression analysis. For perinatal outcomes, we calculated gestational age based on the date of oocyte pick-up (OPU) and in sensitivity analyses on data from Denmark and Norway, we also calculated gestational age based on the second-trimester ultrasonography (US) scan. Risk of pregnancies with same-sex twins after SET was used as a proxy for risk of monozygotic twins. Adjustments were made for child's sex, birth year, parity (0 or >1), maternal age, body mass index, smoking, educational level, fertilization method (IVF/ICSI), the number of aspirated oocytes, SET and country. Information on educational level and the number of aspirated oocytes was not available for Norway. Children born after frozen embryo transfer were not included. The birth cohorts were restricted according to the year in which BT was introduced in the different countries. A higher risk of placenta previa was found in singleton pregnancies after BT compared with CT (adjusted odds ratio [aOR] 2.11 [95% CI 1.76; 2.52]). Singletons born after BT had a higher risk of PTB (aOR 1.14 [95% CI 1.01; 1.29]) compared with CT singletons, when estimated based on OPU. Furthermore, an altered male/female ratio (aOR 1.13 [95% CI 1.06; 1.21]) with more males following BT compared with CT was seen. Risk of same-sex twins after SET was higher after single BT compared with single CT (aOR 1.94 [95% CI 1.42; 2.60]). Residual confounding cannot be excluded, in particular related to duration and cause of infertility that we could not adjust for due to lack of reliable data. Extended embryo culture to the blastocyst stage has the potential to compromise obstetric and perinatal outcomes in fresh cycles. These results are important since an increasing number of IVF/ICSI treatments are performed as BT. NORDFORSK (project no: 71450). The Research Fund of Rigshospitalet, Copenhagen University Hospital. ReproUnion Collaborative study, co-financed by the European Union, Interreg V ÖKS. Grants from Swedish state under the agreement between the Swedish government and the county councils, the ALF-agreement (LUA/ALF 70940), Hjalmar Svensson Research Foundation. The Research Council of Norway through its Centres of Excellence funding scheme, project number 262700. None of the authors has any conflicts of interests to declare regarding this study. ISRCTN11780826.

GM-CSF does not rescue poor-quality embryos: secondary analysis of a randomized controlled trial

PurposeTo evaluate implantation potential of cleavage-stage embryos cultured in medium containing 2 ng/ml granulocyte–macrophage colony-stimulating factor (GM-CSF) versus control medium, according to embryo morphological quality and then transferred on day 3.MethodsExplorative secondary data analysis of a multicenter, randomized, placebo-controlled, double-blinded prospective study of 1149 couples with embryo transfer after IVF/ICSI. This analysis includes a subgroup of 422 subjects with either single-embryo transfer (SET, N = 286) or double-embryo transfer of two embryos with equivalent morphological quality (DET, N = 136). Implantation rate and live birth rate were assessed according to category of morphological embryo quality on day 3.ResultsCulture with GM-CSF did not increase the implantation rate for embryos classified as poor quality. A trend towards greater benefit of GM-CSF on implantation and survival until live birth for top-quality embryos (TQEs) compared with poor-quality embryos was observed, although not statistically significant. For TQEs, the percentage of transferred embryos resulting in a live born baby was: 40.9 ± 5.3% (GM-CSF) versus 30.5 ± 4.6% (control) (P = 0.24; odds ratio [OR] 1.43, 95% confidence interval [CI] 0.79–2.59), and for embryos with less than 6 cells at day 3 this same rate was: 7.4 ± 3.3% (GM-CSF) versus 12.0 ± 4.0% (control) (P = 0.26; OR 0.53, 95% CI 0.17–1.61).ConclusionThis exploratory analysis is consistent with GM-CSF protecting morphologically normal embryos from culture-induced stress and does not support an effect of GM-CSF in rescuing poor-quality embryos.ClinicalTrials.gov identifier: NCT00565747.

Closed vs. Open Oocyte Vitrification Methods Are Equally Effective for Blastocyst Embryo Transfers: Prospective Study from a Sibling Oocyte Donation Program

Purpose: To assess whether open and closed vitrification protocols are equally effective for sibling-oocyte cycles when performing blastocyst embryo transfers. Materials and Methods: A prospective study was set up comparing the open and the closed vitrification techniques in oocyte recipients sharing sibling oocytes between 2014 and 2016. Sibling oocytes were randomly and equally assigned into the closed group (oocytes vitrified in a closed system) or the open group (oocytes vitrified in an open system). Intracytoplasmic sperm injection was performed on all cases. Embryo transfers were performed on day 5. Power analysis calculation showed that 94 cycles would be needed for each group in the study in order to achieve statistical significance at a 5% level with power 80%. Results: The final number of donors included was 95. A total of 190 recipients matched with their donors were included in the study. There was no difference in the mean number of oocytes vitrified with the closed or the open system (8.26 ± 2.54 vs. 8.31 ± 2.57). No significant difference was observed between the 2 groups regarding survival rate, fertilization rate, cleavage rate, top-quality embryos on day 3, blastocyst rate, and top-quality blastocyst rate. Moreover, no statistically significant difference in the b-human chorionic gonadotropin-positive rate, clinical pregnancy rate per cycle, implantation rate, ongoing pregnancy rate, and live birth rate between closed and open groups. Conclusion: Οpen and closed vitrification protocols are equally effective for sibling-oocyte cycles.

The impact of thyroid autoantibodies on the cycle outcome and embryo quality in women undergoing intracytoplasmic sperm injection

BackgroundThe influence of thyroid autoimmunity in in–vitro fertilization/intracytoplasmic sperm injection (ICSI) cycles is still a controversial issue. Although some women are euthyroid, some studies found thyroid dysfunction and thyroid autoimmunity increased the incidence of infertility. We aimed to evaluate the impact of anti-thyroid antibodies on the cycle parameters, embryo quality, and pregnancy outcomes in women who underwent ICSI cycles.ResultsFertilization rate was significantly higher in anti-thyroid antibody-negative patients than in anti-thyroid antibody-positive patients (97.1 ± 10.5 vs. 91.5 ± 19.8, respectively, p = 0.003). Clinical pregnancy, miscarriage, and ongoing pregnancy rates were not different among study groups. The presence of top-quality embryo was lower in anti-thyroid antibody-positive patients but it did not reach any statistical significance (56.5% vs. 67.8%, p = 0.09). In the subgroup analysis, there were no significant differences except for the miscarriage rate according to the TSH threshold.ConclusionThe clinical pregnancy and ongoing pregnancy were not affected by the positivity of anti-thyroid antibodies. We showed that thyroid autoimmunity may have an impact on fertilization rate and embryo quality in patients undergoing ICSI cycles. Miscarriage rate also increases with thyroid autoimmunity and TSH levels above 2.5 μIU/L.

Does the presence of AGG interruptions within the CGG repeat tract have a protective effect on the fertility phenotype of female FMR1 premutation carriers?

While FMR1 premutation carriers (CGG 55-200) were shown to have reduced success with IVF treatment (lower oocyte yield), studies reporting on the association between the number of CGG repeats and patients' response to controlled ovarian hyperstimulation (COH) are inconsistent. In the present study, we aim to explore whether the number of CGG repeats in women with premutation in FMR1 gene, undergoing COH for IVF, correlates with COH variables and whether the number of AGG interruptions may function as a "protective factor" by improving the ovarian response to COH. Retrospective study, in an academic IVF-PGD unit. Fifty-seven consecutive FMR1 premutation carriers who underwent 285 IVF treatment cycles were included. The numbers of CGG repeats and AGG interruptions were retrieved and correlated to the demographics and COH variables. There were no significant association between the numbers of CGG or the AGG interruptions and the number of oocyte retrieved or the peak estradiol levels. The lack of association was also observed when including all the IVF treatment cycles or only the first or last IVF treatment cycle. Moreover, no associations were found between the number of CGG repeats or AGG interruptions and other COH variables, i.e., duration of stimulation, the total dose of gonadotropin used, or the number of top-quality embryos. No associations were observed between the number of CGG repeats or AGG interruptions and any of the COH variables. Further studies are required to identify early biomarkers of POI to empower FMR1 premutation carriers with risk assessment tools to consider procedures such as fertility preservation.

The association between follicle size and oocyte development as a function of final follicular maturation triggering.

To study the association between follicle size and oocyte/embryo quality, as a function of different triggering modes for final follicular maturation. Cohort study conducted in a single tertiary medical centre between July 2018 and May 2019. All women undergoing ovarian stimulation with triggering using human chorionic gonadotrophin (HCG), gonadotrophin-releasing hormone (GnRH) agonist or dual trigger (GnRHa + HCG) were included. Before ultrasound-guided follicular aspiration, follicles were measured and divided into three groups according to maximum dimensions: large ≥16 mm, medium 13-15 mm and small <13 mm. Microscopic examination of the follicular aspirates was performed by an embryologist. Each follicle aspirated was evaluated for oocyte maturation, oocyte fertilization and embryo quality. A total of 640 follicles were measured, including 402 (62.8%) in the large, 148 (23.1%) in the medium and 90 (14.1%) in the small groups. Oocytes were obtained during aspiration from 76.3%, 70.3% and 55.6% of the large, medium and small follicle groups, respectively (P = 0.001). The mature oocyte (metaphase II) rate was significantly higher in the large (P = 0.001) and medium (P = 0.01) compared with the small follicle group. Nevertheless, no between-group differences were observed in fertilization or top quality embryo rates among mature oocytes regardless of the size of the follicle from which they originated. Triggering mode did not influence oocyte recovery rate in the different follicle size groups. A higher oocyte recovery rate was observed from follicles >13 mm, however, mature oocytes achieved similar fertilization and top quality embryo rates regardless of follicle size. Triggering mode did not influence oocyte recovery rate.

Chromosomal polymorphisms are associated with blastomere multinucleation in IVF/ICSI cycles

To investigate the association of chromosomal polymorphisms with multinucleated embryos in infertile couples undergoing in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI). This retrospective case-control study was conducted among 1145 infertile couples undergoing their first IVF/ICSI cycles. According to their karyotype, the couples were divided into chromosomal polymorphism group and control group, and the former group was divided into 3 subgroups: inversion group, D and G genome polymorphic group and 1, 9, and 16 qh+group. The blastomere multinucleation rate, clinical pregnancy rate and live birth rate were compared between the groups. Of the total of 1145 couples, 139 (6.10%) had chromosomal polymorphisms at least in one partner. No significant differences were found in female age, BMI, basal FSH level, total gonadotropin dose, E2 level on day of HCG, number of oocytes retrieved, fertilization rate, top quality embryo rate, clinical pregnancy rate or live birth rate among the groups (P > 0.05). The multinuclear rate of the embryos in couples with pericentric inversion of chromosomes 1, 9, and Y chromosomes and those with D and G genome polymorphisms were 8.23% and 4.65%, respectively, significantly higher than that in the control group (2.69%; P < 0.05); the multinuclear rate of the embryos was 2.77% in 1, 9, and 16 qh+ group, similar with that in the control group (P > 0.05). Infertile couples with pericentric inversion of chromosomes 1, 9, and Y chromosomes and in those with D and G genome polymorphism are at higher risks of blastomere multinucleation in IVF- ICSI cycles; 1, 9, and 16 qh + polymorphisms do not increase the rate of blastomere multinucleation of the embryos.

Recurrent implantation failure: which patients benefit from endometrial scratching prior to IVF?

Endometrial scratching (ES) using a biopsy catheter prior to the IVF cycle in the repeated implantation failure (RIF) population has been suggested, but no convincing evidence of its benefit has been presented until now. A retrospective mono-center study among 300 consecutive IVF-RIF cycles following evaluation of the ovarian reserve, hysterosalpingography or hysteroscopy, pelvic ultrasound, thrombophilia evaluation, karyotyping and assessment of male sperm parametrs. The findings within normal limits. All the patients offered ES, 78 consented and underwent ES prior to their next IVF cycle. A comparison of treatment outcomes between the post-ES cycles (n = 78) and the non-ES cycles (222) demonstrated the following: 34 (43.5%) versus 14 (6.3%) conceptions, respectively (p = 0.001) and 30 (38.4%) versus 2 (0.9%) clinical pregnancies, respectively (p < 0.001%), emphasizing an extremely high biochemical pregnancy rate among the non-ES cycles. Implantation rate was 19.7% versus 0.4%, respectively (p < 0.001) and live birth rate was 33.33% (26 newborns) versus 0.45% (1 newborn), respectively (p < 0.001). Since there were more embryos available for transfer and more top-quality embryos in the post-ES-IVF conception cycles, the role of ES became questionable. A multivariate analysis that included ES and the percentage of top-quality embryos demonstrated that ES was an independent factor highly correlated with conception in this particular RIF population. ES proved to be an efficient tool in a particular subgroup of RIF patients with fertility investigation results within normal limits, an optimal ovarian response to gonadotropins, and a high percentage of top-quality embryos. Nevertheless, the results should not be overestimated, since the study has limitations related to its retrospective model.

ICSI in late matured oocytes, is it worth it? Study with laboratory, clinical and genetic evaluation results.

ObjectiveTo compare laboratory results of embryo development from late matured oocytes in relation to mature oocytes in D+0.MethodsWe carried out a cross-sectional study during the period from January to December 2018, in which we collected data through medical records analysis. 913 oocytes were collected and divided into 3 groups: group 1 - 643 MII oocytes; group 2 - 119 MI oocytes and; group 3 - 151 PI oocytes. These studied oocytes were from different maternal ages and infertility factors. The analyzed variables were fertilization rate, embryo cleavage, top quality embryos on the third day of development, blastocyst stage, top quality blastocysts, euploid blastocysts, top quality blastocysts and gestation. We documented the data, and performed the statistical analysis using the chi-square test (p<0.05).ResultsAll MII oocytes were injected (643); 103/119 MI oocytes and 88/151 PI oocytes that matured late in D + 1, were also injected. The fertilization rate of the three groups did not present statistical difference. The oocytes of group 1 had a statistically proven better prognosis than oocytes from groups 2 and 3 when compared, respectively, embryo cleavage (p=0.000), top quality embryos on the third day of development (p=0.000) and blastocyst formation rate (p=0.004). In the LMO group, there were no euploid embryos and, therefore, there no embryo transfer.ConclusionAlthough late matured oocytes have made blastocyst formation possible, even if in low rates, there were no viable embryos for transfer.

Trimethylamine-N-oxide is present in human follicular fluid and is a negative predictor of embryo quality.

STUDY QUESTIONAre levels of trimethylamine-N-oxide (TMAO) in human follicular fluid (FF) related to IVF outcomes?SUMMARY ANSWERHigher levels of TMAO are a negative predictor of oocyte fertilization and embryo quality.WHAT IS KNOWN ALREADYTMAO is a metabolic product of dietary choline and l-carnitine produced via subsequent enzymatic modifications by the intestinal microbiota and hepatocytes. TMAO promotes inflammatory and oxidative stress pathways and has been characterized as a causative biomarker for the development of cardiometabolic disease.STUDY DESIGN, SIZE, DURATIONFor the present cross-sectional study, samples (FF and plasma) from 431 modified natural cycle (MNC)-IVF cycles of 132 patients were collected prospectively between October 2014 and March 2018 in a single academic medical center.PARTICIPANTS/MATERIALS, SETTING, METHODSTMAO and its precursors (choline, l-carnitine and gamma-butyrobetaine) were measured by ultra-high-performance liquid chromatography/mass spectrometry in (i) matched FF and plasma from 63 MNC-IVF cycles, in order to compare metabolite levels in the two matrices and (ii) FF from 232 MNC-IVF cycles in which only one oocyte was retrieved at follicular puncture. The association between metabolite levels and oocyte fertilization, embryo fragmentation percentage, embryo quality and the occurrence of pregnancy was analyzed using multilevel generalized estimating equations with adjustment for patient and cycle characteristics.MAIN RESULTS AND THE ROLE OF CHANCEThe level of choline was higher in FF as compared to matched plasma (P < 0.001). Conversely, the levels of TMAO and gamma-butyrobetaine were lower in FF as compared to plasma (P = 0.001 and P = 0.075, respectively). For all metabolites, there was a positive correlation between FF and plasma levels. Finally, levels of TMAO and its gut-derived precursor gamma-butyrobetaine were lower in FF from oocytes that underwent normal fertilization (TMAO: odds ratio [OR] 0.66 [0.49–0.90], P = 0.008 per 1.0-μmol/L increase; gamma-butyrobetaine: OR 0.77 [0.60–1.00], P = 0.047 per 0.1-μmol/L increase) and developed into top-quality embryos (TMAO: OR 0.56 [0.42–0.76], P < 0.001 per 1.0-μmol/L increase; gamma-butyrobetaine: OR 0.79 [0.62–1.00], P = 0.050 per 0.1-μmol/L increase) than in FF from oocytes of suboptimal development.LIMITATIONS, REASONS FOR CAUTIONThe individual contributions of diet, gut bacteria and liver to the metabolite pools have not been quantified in this analysis.WIDER IMPLICATIONS OF THE FINDINGSMore research on the contribution of diet and the effect of gut bacteria on FF TMAO is warranted. Since TMAO integrates diet, microbiota and genetic setup of the person, our results indicate potential important clinical implications for its use as biomarker for lifestyle interventions to improve fertility.STUDY FUNDING/COMPETING INTEREST(S)No external funding was received for this project. The Department of Obstetrics and Gynecology of the University Medical Center Groningen received an unrestricted educational grant of Ferring Pharmaceutical BV, the Netherlands. The authors have no other conflicts of interest.TRIAL REGISTRATION NUMBERNetherlands Trial Register number NTR4409.

Do we need to measure progesterone in oocyte donation cycles? A retrospective analysis evaluating cumulative live birth rates and embryo quality.

Does late follicular-phase elevated serum progesterone (LFEP) during ovarian stimulation for oocyte donation have an impact on embryo quality (EQ) and cumulative live birth rate (CLBR)? LFEP does not have an influence on EQ nor CLBR in oocyte donation cycles. Ovarian stimulation promotes the production of progesterone (P) which, when elevated during the follicular phase, has been demonstrated to have a deleterious effect in autologous fresh IVF outcomes. While there is robust evidence that this elevation results in impaired endometrial receptivity, the impact on EQ remains a matter of debate. The oocyte donation model is an excellent tool to assess the effects of LFEP on EQ from those on endometrium receptivity separately. Previous studies in oocyte donation cycles investigating the influence of elevated P on pregnancy outcomes in oocyte recipients showed conflicting results. This is a retrospective analysis including all GnRH antagonist down-regulated cycles for fresh oocyte donation taking place in a tertiary referral university hospital between 2010 and 2017. A total of 397 fresh donor-recipient cycles were included. Each donor was included only once in the analysis and could be associated to a single recipient. The sample was stratified according to serum P levels of ≤1.5 and >1.5ng/mL on the day of ovulation triggering. The primary endpoint of the study was the top-quality embryo rate on Day 3, and the secondary outcome measure was CLBR defined as a live-born delivery beyond 24weeks. Three hundred ninety-seven fresh oocyte donation cycles were included in the analysis, of which 314 (79%) had a serum P ≤ 1.5ng/mL and 83 (20.9%) had a serum P > 1.5ng/mL. The average age of the oocyte donors was 31.4 ± 4.7 and 29.9 ± 4.5years, respectively, for normal and elevated P (P = 0.017). The mean number of oocytes retrieved was significantly higher in the elevated P group with 16.6 ± 10.6 vs 11.5 ± 6.9 in the P ≤ 1.5 group (P < 0.001).In parallel, the total number of embryos on Day 3, as well as the number of good-quality embryos at this stage, was significantly higher in the elevated P group (6.6 ± 5.6 vs 4.15 ± 3.5 and 8.7 ± 6.3 vs 6.1 ± 4.4; respectively, P < 0.001). However, maturation and fertilization rates did not vary significantly between the two study groups and neither did the top- and good-quality embryo rate and the embryo utilization rate, all evaluated on Day 3 (P = 0.384, P = 0.405 and P = 0.645, respectively). A multivariable regression analysis accounting for P groups, age of the donor, number of retrieved oocytes and top-quality embryo rate as potential confounders showed that LFEP negatively influenced neither the top-quality embryo rate nor the CLBR. This is an observational study based on a retrospective data analysis. Better extrapolation of the results could be validated by performing a prospective trial. Furthermore, this study was focused on oocyte donation cycles and hence the results cannot be generalized to the entire infertile population. This is the first study providing evidence that LFEP does not influence CLBR and is adding strong evidence to the existing literature that LFEP does not harm EQ in oocyte donation programs. Not applicable.

Contribution of rescue in-vitro maturation versus double ovarian stimulation in ovarian stimulation cycles of poor-prognosis women.

Maximizing the number of oocytes and embryos obtained in the shortest possible time is of considerable potential clinical significance for women with poor prognosis. The aim of this study was to develop a clinically applicable strategy to obtain more oocytes and viable embryos in one menstrual cycle for poor-prognosis women. Prospective cohort study of 146 women with poor prognosis who received rescue in-vitro maturation (IVM) (n = 50) or double ovarian stimulation (DuoStim) (n = 96) between December 2015 and February 2018. Outcomes, number of oocytes retrieved and matured, and embryo developmental potential were compared between the two groups. The rates of mature oocytes, available embryos and top-quality embryos from luteal phase stimulation (LPS) of DuoStim were all significantly higher than those derived from the immature oocytes of rescue IVM (P < 0.05). The relative contributions of LPS in the DuoStim group for proportion of mature oocytes, available embryos and top-quality embryos were all significantly higher than IVM in the rescue IVM group (P < 0.001). The overall cancellation rate of no oocyte or available embryo significantly decreased from 30.21% to 9.38% (P < 0.001) when DuoStim was carried out, which decreased from 24.00% to 12.00% with no significant difference in the rescue IVM group when immature sibling oocytes were matured in vitro. Rescue IVM and DuoStim can contribute more competent oocytes and viable embryos in the shortest possible time for poor-prognosis women, of which DuoStim may be more efficient.

Increasing paternal age and ejaculatory abstinence length negatively influence the intracytoplasmic sperm injection outcomes from egg-sharing donation cycles.

The paternal role in embryogenesis is much more than providing a haploid genome. The spermatozoa have crucial roles related to embryogenesis' epigenetic regulation, fusion of gametes, and cleavage, which may influence intracytoplasmic sperm injection outcomes. To study whether paternal age, length of ejaculatory abstinence and semen parameters influence the outcomes of intracytoplasmic sperm injection in the oocyte recipients from egg-sharing donation cycles. This historical cohort study involved 427 oocyte recipient intracytoplasmic sperm injection cycles, from 321 patients undergoing egg-sharing donation cycles from January 2015 to May 2017, in a private in vitro fertilization center. Electronic medical records of donors and recipients intracytoplasmic sperm injection cycles were reviewed. The impact of male age, length of ejaculatory abstinence, and semen parameters on recipients' intracytoplasmic sperm injection outcomes were investigated using general mixed models. Implantation, pregnancy, and live birth rates were the primary outcome measures. Paternal age negatively affected fertilization, day 3 top-quality embryos, day 3 normal embryo cleavage speed, blastocyst formation, high-quality blastocysts, and implantation, and was correlated with diminished pregnancy and live birth rates. Length of ejaculatory abstinence negatively influenced the rates of day 3 top-quality embryos, normal embryo development on day 3, blastocyst development, and implantation. Sperm count positively affected the rates of fertilization, blastocyst development, and implantation. The percentage of progressive sperm motility positively affected day 3 normal embryo development rate and implantation rate. Total motile sperm count positively affected blastocyst development rate and implantation rate. Additional tracking of the influence of male characteristics on intracytoplasmic sperm injection outcomes should be stimulated. Paternal age is not manageable, but reducing length of ejaculatory abstinence could be an alternative approach to improve the outcomes of intracytoplasmic sperm injection.