Journal of Assisted Reproduction and Genetics最新文献

Purpose: To evaluate the effects of indirect co-culture of fresh or vitrified ovarian fragments (OFs) with Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs).

Methods: The ovarian fragments were divided into two groups: fresh and vitrified. Some fresh OFs were immediately fixed (fresh control), while others were cultured in vitro for 14 days, either without (monoculture) or with (co-culture) WJ-MSCs. After vitrification and warming, some OFs were immediately fixed (vitrified control), while others were cultured under the same conditions as the fresh OFs. Morphological analysis (classical histology), proliferation (PCNA), senescence (Sudan Black B), expression of genes related to folliculogenesis (FSH-R, LHX8, NANOS3, and FOXL2), apoptosis and anti-apoptosis (BAX and BCL2), and DNA fragmentation (TUNEL) were evaluated in all OFs. Estradiol (E2) levels were measured in the culture medium on days 2, 8, and 14.

Results: The follicular morphology of fresh or vitrified OFs cultured with WJ-MSCs was similar to that of fresh and vitrified OFs fixed on day 0. Regarding development, a higher proportion of normal primary follicles was observed in fresh and vitrified ovarian fragments co-cultured with WJ-MSCs. The presence of WJ-MSCs increased the proportion of stromal cells and proliferative follicles in fresh OFs, and reduced the number of senescent stromal cells in vitrified OFs. Additionally, in fresh OF, the absence of WJ-MSCs reduced BCL2 expression, while their presence in vitrified OF reduced E₂ secretion only at day 14.

Conclusion: JW-MSCs provide a favorable microenvironment for the survival and development of pre-antral follicles cultured in vitro.

Purpose: Advances in assisted reproductive technologies (ART) have led to an increasing number of surplus embryos being cryopreserved. Deciding the fate of these embryos poses complex emotional, ethical, and practical challenges. Understanding the factors that influence these decisions is critical in improving patient counseling and ART policy. This study aimed to analyze patterns in embryo disposition decisions across 6 years in a Portuguese ART center, focusing on differences by family type, patient age, and temporal trends.

Methods: We conducted a retrospective quantitative analysis of embryo disposition forms completed between 2017 and 2022, covering 3494 patients. Data were categorized by year, relationship group (different-sex couples, female couples, and single women), and final decision.

Results: Disposition decisions vary significantly by relationship group. Female couples were more likely to donate embryos to others (31.8%, p = 0.007) and less likely to discard them (15.9%, p = 0.03) than different-sex couples, who had the highest discard rate (26.5%). Single women showed intermediate behaviors. Age was a significant factor: those who donated embryos, particularly to others, were older (mean = 40.6 years) than those who kept or discarded them. No significant differences were found in relation to the number of embryos stored. Temporal analysis showed variation across years, with a notable increase in donations to research in recent years.

Conclusions: Family type and age are important factors influencing embryo disposition decisions, while the number of embryos appears to have a limited impact.

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.

Purpose: Early embryonic arrest (EEA) and implantation failure are prevalent causes of female infertility, with genetic factors playing a significant role. MEI1 is a crucial gene for meiotic chromosome synapsis and is essential for the formation of double-strand breaks (DSBs) during germ cell meiosis. MEI1 variants contribute to EEA and implantation failure. In this study, we reported newly identified mutations in MEI1 that broaden the genetic spectrum associated with embryonic abnormalities.

Methods: Female patients with primary infertility characterized by EEA and implantation failure were recruited, and peripheral blood samples were collected. Whole-exome sequencing was performed, and the identified variants were confirmed by Sanger sequencing. Bioinformatics tools were used to predict the pathogenicity of these variants. Wild-type and mutant plasmids were constructed in vitro and transfected into HEK293T cells. The effects of the variants on the expression and function of MEI1 were investigated via real-time quantitative PCR, western blotting, and immunofluorescence assays.

Results: We identified a novel homozygous variant and a compound heterozygous variant in MEI1 from two unrelated families. Analysis using multiple bioinformatics tools revealed that these variants are rare and may be deleterious. In vitro experiments revealed that these mutations do not alter the subcellular localization of MEI1 but significantly affect its mRNA and protein expression levels, potentially leading to impaired protein function.

Conclusion: These biallelic variants in MEI1 were associated with EEA and implantation failure. Our findings expand the known mutation spectrum of MEI1 and provide further evidence supporting the causal relationship between MEI1 variants and female infertility.