Journal of Assisted Reproduction and Genetics最新文献

Purpose: The current study aimed to investigate whether cathepsin B (cath B) inhibition in cumulus cells of morphologically graded buffalo cumulus-oocyte complexes (COCs) during in vitro maturation (IVM) could modulate apoptotic markers and enhance developmental potential.

Methods: Cumulus cells from grade A and B buffalo COCs were collected at 0, 12, and 24 h of IVM, with or without supplementation of the optimized cysteine protease inhibitor E-64. Cathepsin B and selected apoptotic regulators were quantified by qRT-PCR. Cathepsin B protein levels and apoptotic signals were assessed by immunostaining and TUNEL assay, respectively. Following 24 h of IVM, in vitro fertilization (IVF) and in vitro culture (IVC) were performed, and developmental competence was evaluated based on oocyte maturation, cleavage rate, blastocyst formation rate, and blastocyst quality.

Results: Expression of cath B and pro-apoptotic genes (BID, BCL2, BAX, caspase-3) was found higher along with lower expression of anti-apoptotic genes (BCL2 and XIAP) in the cumulus cells of B grade COCs. The addition of 10 µM E-64 significantly (P < 0.05) reduced cath B and pro-apoptotic gene expression, lowered the BAX/BCL2 ratio, and decreased apoptosis signals in cumulus cells of both COC grades. E-64 supplementation improved blastocyst yield from grade A COCs and enhanced blastocyst quality in both grades, without affecting maturation and cleavage rates.

Conclusion: E-64 supplementation during IVM modulates apoptotic markers in cumulus cells of different grades of buffalo COCs by inhibiting cath B, thereby enhancing the developmental competence and offering valuable insights for improving in vitro embryo production programs in this species.

Purpose: To assess whether the endometrial receptivity analysis (ERA) captures receptivity changes attributed to endometrial aging and whether it may be useful for older patients undergoing fertility treatment.

Methods: Retrospective cohort study of patients who underwent ERA testing at an academic center (01/2019-05/2024). The ERA inferred transcriptomic levels of canonical receptivity markers from biopsies obtained at standard timing. Demographic and treatment-related variables were analyzed by age group. The proportion of non-receptive ERA results (pre- and post-receptive combined) was compared using Fisher's exact test. Univariable and multivariable logistic regression assessed associations between predictors and non-receptive ERA.

Results: Of 210 patients, 205 were included. Age distribution was < 35 (n = 35, 17%), 35-37 (n = 58, 28.3%), 38-40 (n = 53, 25.8%), and ≥ 41 (n = 59, 28.8%). Overall, 166 (81.0%) ERAs were receptive, 33 (16.1%) pre-receptive, and 6 (2.9%) post-receptive. BMI, infertility diagnosis, and prior implantation or miscarriage history did not differ by age. Non-receptive ERA proportions were 20% (< 35), 17.2% (35-37), 17.0% (38-40), and 22.0% (≥ 41) (p = 0.52). In multivariable analysis adjusting for BMI and number of prior failed euploid transfers, age was not associated with non-receptive ERA (aOR 0.98, 95% CI 0.34-2.30, p = 0.97).

Conclusion: Uterine age was not associated with increased odds of non-receptive ERA, suggesting that the test does not capture age-related changes in endometrial receptivity. Although endometrial aging is implicated in reduced embryo transfer success, the ERA should not be ordered solely on the basis of uterine age. The ERA may not reliably detect age-related endometrial differences in the window of implantation at a clinical level.

Purpose: To compare dual triggering versus hCG-only triggering in poor responders undergoing controlled ovarian stimulation with a GnRH antagonist protocol in ART cycles.

Methods: A systematic review and meta-analysis was performed. The primary outcome was the number of mature oocytes (MII). The number of oocytes retrieved, clinical pregnancy, and miscarriage rates were analyzed as secondary outcomes. Subgroup analyses were conducted according to the Bologna and POSEIDON classifications.

Results: Ten studies were included. Dual triggering significantly increased the number of retrieved and mature oocytes in patients classified according to the Bologna criteria, but not in those classified by the POSEIDON criteria. Age-related differences across studies appeared to influence the efficacy of dual triggering. A borderline improvement in clinical pregnancy rates was observed among Bologna-defined patients. Miscarriage rates did not differ significantly between the groups.

Conclusions: Dual triggering appears to improve oocyte yield and maturity in Bologna-defined patients, but this effect is unlikely to apply uniformly across all phenotypes. Persistent heterogeneity in poor responder definitions limits the understanding of the benefits of dual triggering in this population. Well-designed prospective studies with rigorous phenotypic stratification are warranted to identify which patients are most likely to benefit from dual triggering.

Purpose: To explore genetic basis leading to meiotic disruption in human gametogenesis via exome sequencing.

Methods: This study included three consanguineous families with well-defined infertility phenotypes. Exome sequencing was performed for the index case in family 1 and for the trio (index with parents) in the other two families. Sanger sequencing was used for confirmation and family segregation analysis.

Results: Exome sequencing revealed homozygous loss-of-function variations in SPIDR, TOP6BL, and RAD51AP2 in families 1, 2, and 3, respectively. Segregation in individual families revealed that the parents were carriers, as were the fertile siblings in families 1 and 2. All three genes function in double-strand break formation or repair, identified variants may therefore impair, potentially preventing its completion and contributing to infertility in the index cases. Gene-disease relationships (GDR) were re-evaluated due to the addition of new patients and/or variants in the literature.

Conclusion: Our findings provide additional evidence for the role of SPIDR, TOP6BL, and RAD51AP2 as genetic contributors to human infertility due to meiotic errors. For patients with a similar phenotype, genetic screening could be recommended, and the identification of pathogenic variations might help avoid unsuccessful fertility treatments. Additionally, in patients with molecular defects in DNA repair genes, chromosomal instability may increase the risk of cancer; therefore, long-term follow-up by a multidisciplinary team is recommended.

Purpose: Sickle-cell disease (SCD) is a severe autosomal recessive disorder. At-risk couples may prevent transmission either through prenatal diagnosis with possible termination of pregnancy or preimplantation genetic testing for monogenic disease (PGT-M). Data on PGT-M outcomes in this population remain scarce.

Methods: We conducted a monocentric retrospective study (2006-2021). To assess ovarian response to stimulation, each PGT-M cycle for SCD was matched with two control cycles.

Results: Sixty couples underwent at least one ovarian stimulation for PGT procedure for SCD. Eight couples (13.3%) had one affected partner (S/S or S/C) and one carrier (A/S), while 52 couples (86.7%) were both carriers (A/S). Thirty-five couples (58.3%) already had an affected child, and 17 couples (28.3%) requested PGT-M with HLA typing. Median female age at first attempt was 33 years. Overall, 19 couples (31.7%) achieved at least one live birth following fresh or frozen embryo transfer. Among the 17 couples requesting HLA typing, three HLA-matched births (15.7%) and one unmatched healthy birth were achieved. None of the five women affected by SCD achieved a live birth. Ovarian response did not differ significantly between women with sickle cell trait and the controls.

Conclusion: PGT-M is as a viable option for obtaining healthy offspring. These results bolster the argument that PGT-M serves as an alternative to prenatal diagnosis for eligible couples. Our study aims to assist geneticists, gynecologists, and hematologists in providing the necessary guidance before embarking couples on this long and often challenging journey.

Purpose: Can sperm selection through cumulus cells improve embryo quality compared to conventional methods, and is its effectiveness influenced by parental age?

Methods: This prospective clinical trial included 99 ICSI cycles from 95 couples. Sibling oocytes were randomly allocated at the oocyte level to either the study group (cumulus cell-mediated sperm selection after conventional density gradients centrifugation (DGC), 554 oocytes) or the control group (only sperm selection by DGC, 543 oocytes), using a dish designed to facilitate sperm interaction with cumulus cells. The inclusion criteria for this study were patients using their own oocytes, with a medical indication for ICSI, who had at least 6 mature oocytes (MII) in that cycle. For semen samples, inclusion required the ability to adjust the concentration to 10 million/mL. Exclusion criteria included the use of vitrified oocytes, donated oocytes, and semen samples obtained by testicular biopsy or aspiration. Embryo quality was assessed at the blastocyst stage on day 5 according to ASEBIR. A subanalysis evaluated the influence of parental age on outcomes.

Results: The study group showed a significantly higher proportion of good-quality day-5 blastocysts compared to controls (55.2% vs. 45.3%, p = 0.028). No statistically significant differences were observed in overall blastocyst formation or pregnancy rates, although favourable trends were noted. In an age-stratified analysis, a significant improvement in day-5 blastocyst quality among evaluable blastocysts was observed in women aged 40-45 (51.4% vs. 30.4%, p = 0.017), with a non-significant trend toward improved outcomes in men aged 40-53 (44.7% vs. 32.6%, p = 0.083). No differences were seen in younger age groups.

Conclusion: Cumulus cell-mediated sperm selection after DGC using a specialized Oosafe® ICSI Dish with Sperm Selection Channels was associated with an increased proportion of good-quality day 5 blastocysts compared with conventional sperm preparation. While clinical outcomes did not differ significantly, these findings suggest a potential benefit in specific ART subpopulations, particularly those of advanced maternal age. Further adequately powered studies are required to confirm these observations and to determine their impact on clinical outcomes.

The mouse embryo assay (MEA) is the standard test used in assisted reproduction to evaluate the toxicity and effectiveness of culture media and consumables. However, the assay has been criticised for its limited sensitivity, inconsistencies between laboratories, and ethical concerns. Despite the 3Rs principles, over 111 million mice and rats were used in the USA in 2017, with an unknown proportion of these being used in the MEA. While the FDA has provided MEA guidelines, its aim is to phase out animal toxicity testing within 3-5 years. This article explores the possibility of replacing the MEA with the bovine embryo assay (BEA), providing justifications based on ethics, science, practicality, and economics. Through a review of MEA applications, market data, regulatory frameworks and industry disclosures, the article estimates the current impact of the MEA. Incorporating the BEA into regulations could eliminate the need to breed mice for the MEA and greatly reduce the use of animals. Standardising and validating the BEA would provide a reliable and ethically preferable alternative that aligns with the growing demand from regulators and society for non-animal testing methods.

Purpose: To assess the associations between vanishing twin (VT) and fetal reduction (FR) with obstetric and perinatal outcomes following in vitro fertilization (IVF)-frozen embryo transfer (FET).

Methods: This was a retrospective cohort study involving women who had undergone FR or experienced VT during the period from 2012 to 2022. Cohorts were stratified by timing into early (< 15 weeks) and late (≥ 15 weeks) VT/FR. Controls comprised primary singletons and non-reduced twins. The primary outcome measurements were maternal and birth complications.

Results: Among 33,238 ongoing pregnancies, there were 24,316 primary singletons, 7452 non-reduced twins, 1354 VTs, and 116 FRs. Multivariable analyses showed birth outcomes in the study groups were similar to or better than non-reduced twin deliveries. Compared to primary singletons, both early and late FRs were associated with increased risk of preterm birth (PTB); late FR also increased the risk of low birthweight (LBW). Early and late VTs similarly had higher risks of PTB and LBW versus primary singletons. Obstetric complications were generally comparable or lower in the study groups versus twin deliveries; however, late FR was linked to a higher risk of hypertensive disorders of pregnancy compared with primary singletons, and late VT was associated with increased abnormal placentation versus primary singletons and twins.

Conclusions: In this large IVF-FET cohort, most birth and maternal outcomes were comparable or better than in non-reduced twins, but certain complications remained more common in both VT and FR groups. Both exposures were linked to adverse perinatal outcomes versus primary singletons. Moreover, VT and FR appear to be more problematic when these occur later in pregnancy.

Purpose: To evaluate the effectiveness and safety of personalized embryo transfer (pET) guided by TERTs compared with standard embryo transfer (sET) in assisted reproductive technology.

Methods: Systematic review and meta-analysis of randomized controlled trials (RCTs) and cohort studies (CS) at low or moderate risk of bias was conducted. PubMed/MEDLINE, EMBASE, CENTRAL, LILACS, and CINAHL were searched to November 2025 without restrictions. Conference abstracts and reference lists were also screened. Reviewers independently screened, extracted data, and assessed risk of bias. RCTs and CS were pooled separately using random-effects models. Odds ratios (ORs) were synthesized using the generic inverse-variance method. Prespecified subgroups included prior failures and euploid transfers.

Results: We included 44 studies (4 RCTs; 40 CS). Thirty-five studies evaluated ERA, six rsERT, and four other platforms. In women with limited or no prior failures, two low-risk RCTs showed pET with ERA probably results in little or no difference in LBR versus sET (RR 0.98, 95% CI 0.88-1.10; 1069 women; moderate certainty). In women with recurrent implantation failure (RIF) transferring untested embryos, nine low/moderate-risk CS showed a probable increase in LBR with TERT-guided pET (OR 1.58, 95% CI 1.34-1.86; 4754 women; moderate certainty), with similar direction of effect across ERA, rsERT, and ERT. Among RIF women undergoing euploid transfers, five studies provided very uncertain evidence of benefit (OR 1.36, 95% CI 0.83-2.22; 852 women; very low certainty). Findings were heterogeneous and imprecise, yielding very low certainty of evidence.

Conclusion: Current evidence does not support routine use of TERTs in non-RIF. In RIF, TERT-guided pET is probably associated with higher LBR when untested embryos are transferred, but benefits remain uncertain in euploid transfers, reflecting either a small biological effect, methodological bias, or inconsistent protocol implementation. Future research should prioritize adequately powered RCTs in RIF, especially with euploid embryos, and direct comparisons of TERT platforms and assessment of test reproducibility.