Operator-dependent laboratory tasks-embryo selection, vitrification and warming, and intracytoplasmic sperm injection (ICSI)-have been the principal targets of IVF laboratory automation, and a fourth strand of work, integrated end-to-end laboratory automation, has produced its first peer-reviewed clinical evidence in 2026. Clinician-side automation in the same ART cycle has an older, broader literature that provides a useful comparator for what laboratory-side automation has and has not yet achieved. To synthesize the published evidence on automation of these laboratory-side task domains; to distinguish what has been demonstrated in randomized or large multicenter studies from what remains in the proof-of-concept phase; to compare the maturity of laboratory-side automation with the older, broader literature on clinician-side automation; to articulate the structural condition-parallel development of clinician-side and laboratory-side automation-that the available evidence suggests would have to be met before any of these technologies could deliver the system-level outcome gains they promise; and to identify a complementary observation that the laboratory tasks for which automation has been most actively developed are not those in which operator competence translates most directly into clinical outcome, with preimplantation genetic testing (PGT) biopsy and tubing as the conspicuous omission. A narrative review of PubMed-indexed primary studies, ESHRE/Alpha consensus documents, and Cochrane reviews was conducted, with emphasis on randomized comparisons (Hajek 2021 for vitrification, Illingworth 2024 for AI selection) and large registry or multicenter datasets where available. The 2026 Human Reproduction proof-of-concept report on integrated end-to-end laboratory automation (Chavez-Badiola and colleagues) is treated as the current state of evidence for that domain. (i) Deep-learning embryo grading reproducibly outperforms individual embryologists in retrospective benchmarks across multiple algorithms and centers, but the only published randomized trial failed to demonstrate non-inferiority of deep-learning selection over standard morphology in clinical pregnancy, and the Cochrane review of time-lapse imaging found no evidence of differences in live birth or clinical pregnancy. The realistic value proposition is workflow standardization rather than improved clinical outcome. (ii) Closed semi-automated vitrification (Gavi) achieves clinical and survival outcomes equivalent to manual Cryotop in a multicenter RCT; one-step warming protocols offer substantial workflow gains without compromising survival. (iii) Automated and remotely-operated ICSI have produced healthy live births in proof-of-concept clinical work but remain very early in their clinical adoption curve. (iv) Integrated end-to-end laboratory automation has produced its first proof-of-concept clinical evidence in a single sponsor-conducted pilot in 11 selected patients, in which automated arms were numerically outperformed by manual sibling-oocyte controls; randomized comparison and independent replication are not yet available. (v) Across all four domains, the clinical endpoint of an ART cycle is jointly determined by laboratory-side and clinician-side performance; this co-dependence, examined in detail in the "The clinician's perspective: where automation has earned trust and where it has not yet" section, places a structural constraint on the system-level benefit any single laboratory-side technology can be expected to deliver. A within-laboratory parallel of this asymmetry is also observed: automation effort has converged on the technically tractable laboratory tasks (ICSI, vitrification, and embryo selection) rather than on the tasks with the highest operator-competence elasticity (PGT biopsy and tubing). Across the four laboratory-side task domains, the level of clinical evidence available in 2026 is uneven and the maturity gradient is asymmetric with respect to clinician-side automation, which has a longer accumulated evidence base in the same ART cycle. Rather than disappearing, the embryologist's role is shifting toward verification, exception-handling, and quality oversight. The substantive observation a focused review can make in 2026 is that the system-level benefits projected for laboratory-side automation appear bounded by the slower pace of clinician-side automation development and that the asymmetry between the two halves of the cycle, together with a parallel asymmetry inside the laboratory itself, conditions what any laboratory-side technology can be expected to deliver at the level of patient outcome.
Does ovarian reserve, as measured by anti-Müllerian hormone (AMH), influence the utilization of preimplantation genetic testing for aneuploidy (PGT-A) in autologous IVF cycles across different age groups? Lower ovarian reserve, reflected by reduced AMH levels, was associated with a lower likelihood of undergoing PGT-A across all age strata. PGT-A is widely used in IVF, particularly in older patients, but its utilization may be influenced not only by age but also by expected ovarian reserve and embryo yield. How ovarian reserve is associated with selection for PGT-A at a population level has not yet been well characterized. This cross-sectional analysis of the US Society for Assisted Reproductive Technology Clinic Outcome Reporting System (SART CORS) database includes 258,532 patient-first autologous ovarian stimulation cycles performed between 2014 and 2021. The study included the first autologous IVF cycles of women aged 21-46 years. Donor oocyte, donor embryo, and gestational carrier cycles were excluded, as were cycles using <150 IU/day of gonadotropins or with missing key data. Cycles were classified according to PGT-A use. Ovarian reserve was primarily assessed using the most recent AMH value measured within 1 year of treatment. Multivariable logistic regression models evaluated the association between AMH and PGT-A utilization, adjusting for age, calendar year, race, total gonadotropin dose, gravidity, and prior ART, with prespecified testing for the age-AMH interaction. Across the study population, higher AMH levels were associated with significantly increased odds of undergoing PGT-A. This association persisted after multivariable adjustment and within each age group. AMH modelled as both a continuous variable and dichotomized at 1 ng/ml showed consistent results, indicating an independent relationship between ovarian reserve and PGT-A utilization that was unlikely due to chance. The observational, cross-sectional design limits causal inference. Residual confounding is possible, and registry data do not capture treatment intent or all of the clinical reasons underlying use or non-use of PGT-A. Embryo-level genetic outcomes and clinical outcomes such as live birth were not evaluated. These findings suggest a systematic selection pattern in IVF practice, whereby patients with diminished ovarian reserve are less likely to undergo PGT-A. This population imbalance should be considered when interpreting observational studies of PGT-A and when counselling patients about the clinical context in which PGT-A is most commonly used. This work was supported by intramural funds from The Center for Human Reproduction and the not-for-profit research Foundation for Reproductive Medicine, both in New York, NY, USA.Drs. Barad and Gleicher are co-inventors on several US patents hold patents related to androgen treatment in females (DHEA) with numbers US8067400B2, US8501718B2, and US9375436B2 (listed in USPTO/public records and assigned to American Infertility of New York). They also receive royalties from Nutraceuticals LLC, which has helped commercialize DHEA as a nutritional supplement. Dr. Gleicher is also a shareholder in Fertility Nutraceuticals and the owner of the Center for Human Reproduction. Dr. Albertini receives a stipend as Editor-in- Chief of the Journal of Assisted Reproduction and Genetics and receives consulting fees and travel support from Ferring Pharmaceuticals. Drs. Darmon, Gayete-Lafuente, Nicholas, Guijarro-Baude, and Patrizio report no conflicts of interest. N/A.
Machine learning (ML) is increasingly being introduced into assisted reproduction clinical practice, particularly for embryo grading and selection. This study aimed to explore how assisted reproductive technology (ART) professionals, regulators and patients understand the use of ML in embryo assessment. Semi-structured interviews were conducted with ten ART professionals/regulators and ten patients undertaking assisted reproduction in Australian clinics. Interviews explored themes identified as ethically salient to clinical implementation. Both professionals and patients expressed mistrust of ML and emphasised the importance of human oversight in embryo selection. Participants identified uncertainties regarding how ML produces knowledge and raised questions about accountability and responsibility for ML-assisted decisions. Concerns were expressed that ML may lead to the discarding of viable embryos in the pursuit of selecting embryos. Participants called for full disclosure from clinics regarding the use of ML. Most professionals expressed a preference for transparent ML models and raised concerns about incorrect scoring due to poorly trained algorithms, handling errors, and potential algorithmic bias. Some professionals also expressed concern about over-reliance on ML and the potential for workforce deskilling. ART professionals and patients are cautious about the introduction of ML into embryo selection and emphasize the continued importance of human oversight, transparency, and accountability. Knowledge production with ML remains contested and there is uncertainty on how best to incorporate ML and reconfigure the work of embryologists and their professional expertise.
The objective was to study the effect of exposure to virtual reality (VR) on clinical pregnancy rate (CPR) and anxiety levels in patients undergoing frozen embryo transfer (FET), one of the most stressful stages of the in vitro fertilization (IVF) treatment journey. A single-site prospective experimental randomized controlled trial was conducted between 05/2019 and 05/2023. Three hundred fifty FET patients using their own oocytes, or donated oocytes/embryos (oocyte provider's aged 21-45) were enrolled and randomized in a 1:1 ratio to the VR and control groups. Clinical pregnancy rates (CPR) were assessed at 2, 6, and 12 weeks. Stress physiological parameters (heart rate, saliva samples, and systolic/diastolic blood pressure) and anxiety (STAI-State questionnaire) were assessed at baseline (T1), pre-FET (T2) and post-FET (T3). Patients were blinded to their group assignment at T1 and T2 measurements. Pregnancy rates at 2 weeks were higher in the VR group than in the control group (52.6% vs. 44.6%, p = 0.134). The CPR increased significantly at 6 weeks (45.7% vs. 34.9%, p = 0.038) and 12 weeks (42.9% vs. 28.6%, p = 0.005) in the VR compared to the control group. In the multivariate logistic regression analyses, the total number of previous ovum pick-up, quality of the transferred embryo, and receiving VR exposure were the three significant predictors of increased CPR at 2, 6, and 12 weeks. Differences in the physiological stress parameters of all participants at three timepoints were found (p < 0.05) but did not differ between the two groups (p > 0.05). STAI-State anxiety scores among the VR group dropped much more substantially from T2 to T3 than in the control group (mean difference of 5.2 vs. 2.7, p = 0.026). The findings of this large-scale RCT suggest that one-time VR exposure before FET, along with other clinical factors such as the quality of transferred embryos, significantly increased CPR and decreased participants' stress and anxiety levels associated with the FET procedure. Clinical Trial Registry: Clinical Trials.gov. NTC04394962 https://clinicaltrials.gov/study/NCT04394962 Registration Date: 2020-05-06. Date of Initial Enrollment: 2019-05-31.
The accurate identification of gametes, embryos, and patients is a fundamental requirement for the intended treatment in the context of assisted reproductive technology (ART). Embryo mix-ups are among the most serious errors in ART and are considered highly sensitive, which can impede error management and the identification of error sources. Here, we present five cases of embryo mix-ups that occurred at five different ART facilities. These reports highlight misidentification of patients and specimens as underlying factors in all five mix-up cases and emphasize the importance of facility-wide root cause analysis to improve process safety for patients and staff. Although human error was an important factor, the underlying conditions that make these critical errors possible must be identified to allow for effective risk mitigation. In all cases, a sequence of errors led to embryo mix-ups that could have been prevented by adherence to clear institutional guidelines and workflows regarding patient identification and verification of patient and sample identities prior to embryo transfer. It is important that operating guidelines are integrated into daily work routines and ART facilities have control systems in place to monitor critical steps and allow for the detection of error sources or faulty processes to prevent subsequent damage.
Severe asthenoteratozoospermia (ATZ) is a major cause of male infertility and is frequently associated with defects in sperm flagellar architecture. DNAH12 encodes a dynein heavy chain of the inner dynein arm (IDA); however, the spectrum of sperm structural abnormalities associated with DNAH12 mutations in humans remains incompletely characterized. Whole-exome sequencing (WES) was performed in two infertile men with severe ATZ. Sperm from patients and fertile controls were examined by immunofluorescence (IF) staining for DNAH12 and related axonemal proteins, hematoxylin and eosin (H&E) staining for sperm head and tail morphology, and transmission electron microscopy (TEM) for ultrastructural evaluation. The developmental expression pattern of DNAH12 was analyzed using integrated single-cell transcriptomic datasets. Intracytoplasmic sperm injection (ICSI) outcomes were assessed to evaluate reproductive potential. In this study, two novel homozygous loss-of-function (LoF) variants in DNAH12 (c.5442dupT and c.6286C > T) were identified. DNAH12 deficiency in patient sperm was accompanied by loss of DNAH1, DNALI1, RSPH9, and SPAG6 and disruption of the classical "9 + 2" axonemal structure. Although H&E staining and TEM revealed marked abnormalities in both flagellar and head morphology, the acrosomal region and key functional markers of the sperm head remained preserved. Single-cell analyses showed stage-specific DNAH12 expression from secondary spermatocytes to round spermatids, consistent with roles in early flagellar assembly and sperm head morphogenesis. Both patients achieved normal fertilization and embryo development following ICSI. These findings expand the DNAH12-related spectrum of male infertility and support ICSI as an effective reproductive option for affected individuals.
Embryo transfer is a critical final step of IVF treatment. The definition of difficult transfer is subjective and not accurate. One of the characteristic that makes a transfer seem difficult is ''time consuming'', although there is no specific time cutoff identified. Limited studies examined the relationship between transfer time and pregnancy outcomes, with conflicting results. The duration of the whole procedure beginning with the passage of a trial catheter through the internal os, until the transfer is performed, was barely assessed. This study aimed to assess the effect of the duration of the entire transfer process on clinical pregnancy rates. This was a single center retrospective cohort study, including fresh cycles with time recording between 2015 and 2023. The total duration was divided into four quartiles. All transfers were performed by staff physicians, under direct transabdominal ultrasound guidance. The procedure begins with the passage of a trial catheter through the internal os. The inner catheter is removed, and the loaded catheter is then passed through the outer sheath to the desired location within the endometrium. Data collected included basic demographic data, infertility diagnosis and ovarian reserve parameters. The primary outcome was the clinical pregnancy rate per transfer, defined as the presence of a gestational sac and a fetal heartbeat using ultrasonography one month after embryo transfer. Secondary outcomes included live birth rates and miscarriage rates. Statistical analysis included the Fisher-exact test and ANOVA. Multivariate logistic regression was performed to control for relevant confounders. The study included 1079 fresh cycles. The total duration of the transfer ranges from 1-9 min. The study group was divided into four quartiles (minutes): Q1 (1-3.43), Q2 (3.44-4.88), Q3 (4.89-6.46), and Q4 (6.47-9). Female age was younger in Q3 and Q4 compared to Q1 (38.1 ± 5.2 (Q1) vs. 35.5 ± 5.7 (Q3) and 34.8 ± 6 (Q4), P = 0.001). There were no differences in BMI in the four groups (P = 0.31), no differences in cycle rank (P = 0.38), and no differences in gravidity and parity (P = 0.53 and P = 0.39, respectively). In univariate analysis, clinical pregnancy rates per transfer were higher in Q3 compared to Q1 (20.1% in Q1 compared to 31.1% in Q3; P = 0.003). The rate of live birth was higher in Q2 compared to Q1 (23.6% in Q2 vs. 7.7% in Q1, P = 0.003), and miscarriages in Q1 were higher compared to Q2 (40.6% vs. 16.8%, P = 0.002). In multivariate logistic regression, after controlling for relevant confounders, including embryo stage, grade and number of embryos transferred, and female age, longer transfer time was not negatively associated with pregnancy rates. With Q1 as reference, adjusted odds ratios (95% confidence interval) were 0.919 (0.538-1.571), 1.076 (0.631-1.834) and 0.627 (0.353-1.111) for transfer time groups Q2, Q3, and Q4, respectively. A longer transfer time is not negatively associated with clinical pregnancy rates after controlling for relevant confounders. To support our findings, larger prospective studies should be conducted.
Spermatogonial stem cells (SSC) are of high significance in animal reproduction, breeding, and regenerative medicine. Cryopreservation of putative SSC is a prerequisite for long-term storage and future applications. However, dissociated putative SSC are very susceptible to cryostress and undergo apoptosis, thereby reducing their functional competence. To minimize the dissociation-induced apoptosis during cryopreservation and post-thaw culture, the role of anti-apoptotic molecule, Y-27632, was evaluated in sheep putative SSC. The putative SSC were isolated, enriched, and cultured from prepubertal sheep testis. Y-27632 (0, 5,10, and 15 µM) was added in putative SSC cryomedia (Cy group) and in both freezing and post-thaw culture media (Cy + y group). The effects were compared with non-treated control (C group). One month after cryopreservation, the putative SSC were assessed for viability, ROS production, metabolic activity, stemness associated undifferentiated status, colony characteristics, and apoptosis associated pathway/genes. The viability of the dissociated post-thaw putative SSC was significantly higher > 90% when cryopreserved in presence of Y-27632, compared to control (C; 82.5 ± 1.7%). The metabolic activity and stemness associated undifferentiated status were also significantly (p < 0.05) improved in 10 and 15 Cy + y as compared to C. ROCK1 and p53 expression was significantly (p < 0.05) downregulated in 10 Cy + y compared to the C. Also, a significantly (p < 0.01) higher apoptotic cells were observed in C in comparison to 10 Cy and 10 Cy + y (3.9 ± 0.8% vs 1.9 ± 0.2% and 0.35 ± 0.05%). These findings suggest that 10 µM Y-27632, when added to cryomedia and post-thaw culture, reduces apoptosis and thereby improves viability of cells during cryopreservation. It also helps in maintaining undifferentiated status and metabolic activity of sheep putative SSC.
Primary ciliary dyskinesia (PCD) is a rare genetic heterogeneous disorder mainly characterized by impaired mucociliar clearance and chronic respiratory symptoms. Although DNAH5 is commonly implicated in PCD, several DNAH5 variants remain unclassified. The proband was studied by high-speed videomicroscopy, transmission electron microscopy, whole exome sequencing and immunofluorescence. Protein structural analysis was performed through structural models obtained by X-ray crystallography and cryo-electron microscopy. The family was studied by Sanger sequencing of the variants, high-speed videomicroscopy and immunofluorescence. The patient carry, in heterozygosity, the DNAH5 c.5290 T > C p.(Ser1764Pro) missense variant of uncertain significance and the pathogenic truncated variant DNAH5 c.4237C > T p.(Gln1413*). He was clinically diagnosed in adulthood with PCD, confirmed following nasal nitric oxide measurement, high-speed videomicroscopy and transmission electron microscopy, all of which revealed hallmark PCD defects, with decreased nasal nitric oxide levels (30 nl/min) and ciliary beating frequency (0.66 Hz), a dyskinetic ciliary beating pattern (62.5% total immotility) and a class-1 ultrastructure. Immunofluorescence analysis demonstrated reduced DNAH5 expression, and protein structural models predicted that the variant of uncertain significance causes an unstable protein. Family analyses confirmed a trans-inheritance and uncovered a brother with a similar PCD phenotype, the same two variants and similar reduced DNAH5 expression. Results support a pathogenic role for the c.5290 T > C p.(Ser1764Pro) variant and elucidate the effects of the other variant. These results underscore the importance of integrating clinical, ultrastructural, molecular and protein expression analyses to clarify and contribute to PCD diagnosis, besides now serving as potential markers for diagnostics and targeted therapies.
With the widespread adoption of blastocyst culture and advances in genetic testing technologies enabling reliable determination of ploidy status, the traditional concept of normal fertilization is being re-evaluated. Although the presence of two pronuclei (2PN) is conventionally considered indicative of normal fertilization, not all 2PN-derived embryos are euploid. A proportion of trophectoderm (TE) biopsies analyzed by next-generation sequencing (NGS) for preimplantation genetic testing for aneuploidy (PGT-A) are identified as triploid. To determine whether specific morphological and morphokinetic features can differentiate 2PN-derived triploid blastocysts from 2PN-derived euploid blastocysts. Retrospective study including 3369 cycles from 2666 patients aged 18-45 years who underwent PGT-A with NGS and single-nucleotide polymorphism (SNP) genotyping of TE biopsies at our IVF center from 2019 to 2023. Among 12,395 analyzed blastocysts, 59 were classified as triploid, originated from 57 PGT-A cycles and belonged to 54 patients. Fertilization assessment was reevaluated. Triploid embryos were analyzed according to second polar body extrusion, day of biopsy (day 5 vs. day 6), the quality of the TE and the inner cell mass (ICM), and morphokinetic parameters (cleavage times from 2- to 5-cell stage). Categorical variables were expressed as proportions. Student's t-test was performed for continuous variables, with a P-value < 0.05 considered statistically significant. Genetic analysis identified 50 triploid blastocysts derived from 2PN zygotes. Two subpopulation of triploid blastocysts were distinguished: 2 PN 2 polar bodies (PB)-derived and 2PN/1 PB-derived blastocysts. Compared to their euploid sibling embryos, triploid embryos demonstrated (a) significant pronuclear dimorphism, characterized by one PN being significantly larger than the other; (b) delayed morphokinetic parameters, exhibiting slower cell division rates starting from the first cleavage division (t2: 28.59 ± 3.74 h vs. 25.70 ± 2.90 h; t3: 39.76 ± 3.48 vs. 36.75 ± 3.03 h; t4: 40.75 ± 4.07 vs. 37.25 ± 3.66 h; t5: 54.85 ± 8.07 vs. 46.34 ± 5.54 h, respectively; P < 0.05); and (c) reduced embryo quality, with a lower proportion of blastocysts exhibiting high quality TE. The findings primarily support increased awareness of triploidy risk among embryos derived from apparently normal (2PN) fertilization. Morphological and morphokinetic parameters, although associated with triploidy, do not provide sufficient discriminatory accuracy to replace direct genetic assessment of ploidy, which remains the only reliable method to exclude triploidy. Most 2PN-derived triploid embryos appear to result from retention of the second polar body, highlighting the importance of accurate fertilization checks with special consideration given to the extrusion of the second PB. This consideration is especially relevant in non-PGT-A cycles or when the testing platform does not permit ploidy determination, as triploidy is the most common form of aneuploidy observed in first-trimester miscarriages. The incidence of triploidy in 2PN zygotes may vary across IVF laboratory settings. Given the limited number of triploid embryos identified, these findings should be interpreted with caution.
Severe combined immunodeficiency (SCID) is a life-threatening primary immunodeficiency disorder. This study aimed to identify novel recombination activating gene 1 (RAG1) variants in a Chinese pedigree and characterize their impact on protein structure and function, providing a genetic basis for preimplantation genetic testing for monogenic (PGT-M) cycle. Potential RAG1 mutations of the probands were screened by whole-exome sequencing (WES) and confirmed by Sanger sequencing. Configuration predictions of the variants were achieved using SWISS-MODEL. PROVEAN, PolyPhen-2, and MutationTaster were used to predict their pathogenicity. Isogenic pre-B cell lines carrying the mutations were established via CRISPR-Cas9 RNP editing. Functional impacts were assessed through western blotting, proliferation ability, and apoptosis analysis. We identified novel compound heterozygous RAG1 variants c.946T > G (p.C316G) and c.1197_1199del (p.L400del) in two affected siblings with typical SCID. Familial genotyping confirmed autosomal recessive inheritance, with each parent as an asymptomatic carrier of one variant. Both mutations were highly conserved and predicted to be pathogenic. Structural modeling revealed disruption of RAG1 secondary and tertiary structure, affecting zinc-binding (p.C316G) and hydrogen-bonding (p.L400del) interactions. Functional studies demonstrated markedly reduced RAG1 protein expression, synergistic impairment of RAG2 expression, and significantly elevated apoptosis in double-mutant pre‑B cells. Further investigation indicated dysregulation of the PI3K/AKT1/FOXO1 pathway, evidenced by increased phosphorylation of AKT1 and FOXO1. Our study provides genetic and functional evidence that biallelic RAG1 p.C316G and p.L400del mutations act synergistically to cause SCID through protein destabilization, disruption of RAG1/RAG2 complex integrity, and induction of pre‑B cell apoptosis likely mediated by PI3K/AKT1/FOXO1 signaling dysregulation. These findings expand the mutational spectrum of RAG1 and support the clinical application of PGT-M for affected families.
Adolescents and young adults (AYA) who receive gonadotoxic treatment are at risk of future infertility. Fertility preservation is recommended to improve long-term quality of life, but outcomes are less well characterised in AYA than in adults. This study sought to analyse oocyte cryopreservation procedures, outcomes and ethical considerations, to improve counselling for AYA patients and their families. Single-centre, retrospective observational cohort study of AYA patients (14-25 years) referred for fertility counselling and oocyte vitrification prior to gonadotoxic treatment between July 2018 and July 2025. Of 47 referrals received, 37 patients initiated ovarian stimulation, and 36 underwent oocyte cryopreservation. Oncological indications included haematological malignancy (43%), sarcoma (30%) and neurological (9%); benign conditions included aplastic anaemia (6%), neurofibromatosis (2%) and desmoid tumour (2%). The median age of patients who underwent ovarian stimulation was 16 years (range 13-24), with median AMH of 15 pmol/L (range 5.1-81.7 pmol/L). Ultrasound follicle tracking was performed transvaginally in 8/37 (21.6%) and transabdominally in 29/37 (78.4%); transvaginal oocyte collection was performed in 34/36 (94.4%) and laparoscopic in 2/36 (5.63%). Median number of oocytes cryopreserved was 11 (range 0-31), with oocyte maturity rate of 80.5% (range 50-100%). Fertility preservation in post-pubertal AYA patients is feasible, and transabdominal ultrasound monitoring can be used successfully where transvaginal monitoring is not appropriate. Multidisciplinary input is important to ensure fertility preservation can be performed safely, particularly for those with haematological malignancies. To enable clinicians to appropriately counsel young patients, follow-up studies of livebirth outcomes will be important in the long term.
Accurate assessment of oocyte developmental competence remains challenging in IVF, as morphology alone offers limited insight into intrinsic oocyte quality. Emerging evidence indicates that oocyte competence reflects coordinated regulation of the follicular microenvironment rather than a single biomarker. This study aimed to evaluate a multidimensional follicular marker panel integrating oxidative stress parameters, hormonal environment, granulosa cell apoptosis, chromatin integrity and PTEN/PI3K/AKT/mTOR signaling in relation to IVF outcomes. Thirty women undergoing ICSI were included, yielding a total of 211 oocytes. Individual follicles were analyzed according to oocyte maturity, embryo quality and pregnancy outcome. Follicular fluid oxidative stress parameters (TAC, TOC, OSI and MDA) and hormonal mediators (FSH, AMH, melatonin, TGF-β and β-hCG) were assessed. Granulosa cells were evaluated for DNA fragmentation, chromatin integrity and expression of PTEN/PI3K/AKT/mTOR pathway proteins. Favorable IVF outcomes were associated with higher total antioxidant capacity and lower oxidative stress index and malondialdehyde levels, alongside reduced granulosa cell DNA fragmentation and improved chromatin integrity (p < 0.05). These outcomes were characterized by increased PI3K and AKT expression with concomitant mTOR suppression, while PTEN, PDK1, TSC1 and TSC2 levels remained unchanged. Follicular FSH and AMH were primarily linked to oocyte maturity, whereas melatonin, TGF-β and β-hCG were elevated in follicles yielding good-quality embryos and positive pregnancy outcomes (p < 0.05). Oocyte developmental competence in IVF reflects a coordinated follicular microenvironment integrating oxidative balance, granulosa cell integrity, hormonal regulation, and PI3K/AKT/mTOR signaling, supporting a multidimensional, panel-based framework for integrated follicular assessment.
Infertility represents a growing global health challenge, intensifying the demand for advanced assisted reproductive technology (ART). Artificial intelligence (AI) is emerging as a transformative force in reproductive medicine, offering novel solutions to augment clinical success and optimize patient-centered care. This review comprehensively synthesizes AI advancements across the continuum of ART, including sperm and oocyte evaluation, embryo selection, pregnancy prediction, fertility assessment, and supportive nursing. Through the integration of multimodal data, extraction of discriminative features, and construction of predictive models, AI introduces unprecedented objectivity and precision into gamete and embryo analysis, thereby facilitating personalized treatment strategies. Furthermore, intelligent consultation and management systems powered by large language models are redefining reproductive healthcare delivery by enhancing clinician-patient communication and improving engagement. While challenges pertaining to data privacy and model generalizability remain, the deep integration of AI with reproductive medicine is an irreversible trend poised to overcome existing ART bottlenecks and forge a more efficient, humane diagnostic and therapeutic ecosystem.
Which infertility treatment pathway is most cost-effective for women with polycystic ovary syndrome (PCOS)-related infertility in centres with expertise in oocyte in vitro maturation (IVM)? Cost-effectiveness analysis of treatment pathways for PCOS-related infertility using a Markov decision-analytic model. Real-life data from 517 anovulatory PCOS patients treated between January 2018 and January 2023 at a Belgian tertiary infertility clinic informed model parameters and defined the treatment-as-usual (TAU) strategy. Five pathways including incremental cycles of letrozole, low-dose gonadotropins, assisted reproductive technology (ART) after conventional ovarian stimulation (COS) or IVM were modelled and compared to TAU. Patients transitioned between treatment cycles, resulting in ongoing pregnancy or drop-out over a 24-month horizon. Costs were assessed from healthcare and societal perspectives, including direct and indirect costs. Incremental cost-effectiveness ratios (ICERs) were calculated, with sensitivity analyses performed. Ongoing pregnancy rates (OPR) after the first, fourth, and sixth letrozole cycles were 16.1%, 41.6%, and 45.7%, with minimal gain beyond 4 cycles. Deterministic analysis identified two cost-effective pathways: (a) 4 cycles of letrozole followed by 2 cycles of low-dose gonadotropins and COS, and (b) 4 cycles of letrozole followed by 2 cycles of low-dose gonadotropins, 1 cycle of IVM, and COS, with ICERs of -€8174 and -€10,805 from the healthcare perspective, and -€11,494 and -€14,083 from the societal perspective, respectively. Incorporating IVM as second-line would require a 25.7% relative OPR increase from IVM, to become the most cost-effective pathway. Probabilistic sensitivity analyses confirmed robustness. This model highlights the role of IVM as a valuable component of PCOS infertility treatment in centres of expertise, with potential for greater impact as culture systems advance.
To compare live birth rates after in vitro fertilization (IVF) with oocyte donation between women with Turner syndrome and women with premature ovarian insufficiency (POI) of other etiologies and to evaluate the relationships between uterine volume, duration of hormone replacement therapy (HRT), and obstetric outcomes. This retrospective cohort study included 97 women with POI undergoing IVF with oocyte donation at a university-affiliated tertiary care center between 2007 and 2023, including 30 women with Turner syndrome and 67 with POI of other etiologies. Artificial endometrial preparation cycles were used for all recipients. Uterine volume was measured by transvaginal ultrasound prior to the first embryo transfer. Primary outcome was live birth per embryo transfer. Secondary outcomes included live birth according to embryo transfer rank, associations between uterine volume and obstetric or neonatal outcomes, and the relationship between uterine volume and duration of prior HRT. A total of 245 embryo transfers were performed. Live birth rates per embryo transfer were comparable between women with Turner syndrome and those with other POI etiologies (29% vs. 27%). However, women with Turner syndrome had a significantly lower probability of achieving live birth at the first embryo transfer (adjusted odds ratio [OR] 0.09; 95% confidence interval [CI] 0.01-0.64), whereas no significant difference was observed after three or more transfers. Median uterine volume prior to IVF did not differ significantly between groups, despite a substantially longer duration of prior HRT in women with Turner syndrome (15.5 vs. 3.0 years). No significant association was observed between uterine volume and duration of HRT in either group. Among singleton pregnancies, decreasing uterine volume was associated with increased risks of preeclampsia, intrauterine growth restriction, non-cephalic presentation, and adverse neonatal outcomes. Women with Turner syndrome achieve live birth rates after IVF with oocyte donation comparable to those of women with other POI etiologies, although more embryo transfer attempts may be required. No clear association between uterine volume and duration of HRT was observed. However, decreasing uterine volume was associated with adverse obstetric outcomes, highlighting the importance of early hormonal management and careful obstetric surveillance in this population.
Media representations have long influenced public understandings of infertility and assisted reproductive technologies (ART), with traditional media frequently relying on moralized or sensational narratives that reinforce stigma and oversimplification. The rise of social media and artificial intelligence (AI) has reshaped reproductive health communication, allowing patients to share lived experiences, build community, and access information outside clinical settings, while also increasing exposure to misinformation. This review synthesizes peer-reviewed research, clinical commentary, and digital media analyses to examine infertility narratives, online patient engagement, and AI-driven information dissemination. Findings indicate that social media has expanded patient agency and reduced isolation, but AI-generated content and influencer marketing contribute to the rapid spread of inaccurate or misleading fertility information. Despite these risks, digital platforms offer significant opportunities for evidence-based education and empathetic engagement. Clinicians who thoughtfully engage with social media and AI can counter misinformation, direct patients to trustworthy resources, and strengthen patient-physician relationships. When used responsibly, these tools can enhance communication and promote more informed, compassionate infertility care.
To overcome the limited sensitivity of the standard mouse embryo assay (MEA) for embryotoxicity screening of assisted reproduction devices and to assess the bovine embryo assay (BEA) as a suitable alternative. In a large comparative laboratory study, sperm selection, fertilization, and embryo culture media from two suppliers (Vitrolife and Genea Biomedx, respectively) were used to generate bovine blastocysts from cumulus-oocyte complexes obtained from slaughterhouse ovaries and inseminated with frozen semen from the same bull (BEA). Bovine culture media were used for the control group. BEA assessed the following in the three groups: cleavage; blastocyst development and kinetics; post-warming re-expansion/hatching; total cell number; inner cell mass (ICM) and trophectoderm (TE) allocation; and ICM/TE ratio. In parallel, the culture media from both suppliers were tested using mouse cumulus-oocyte complexes that had been inseminated with epididymal sperm (IVF-MEA) and in vivo-derived one-cell embryos (standard MEA). BEA detected differences consistent with reduced developmental competence and embryo quality across 4118 bovine oocytes (≥ 4 independent cycles; ≥ 50 oocytes/group). Sperm selection media performed similarly. Regarding fertilization media, the bovine control yielded higher day 8 blastocyst rates than Vitrolife (P < 0.005), while Genea Biomedx exhibited superior blastocyst quality indicators than Vitrolife (P < 0.05). For embryo culture media, the bovine control outperformed Genea Biomedx, with Vitrolife performing intermediately (P < 0.05). Multiple kinetic, hatching, first lineage allocation and survival endpoints were reduced for Genea Biomedx (P < 0.05). Standard MEA found no significant differences, whereas IVF-MEA (1028 oocytes) detected a difference in day 4 blastocyst yield only. BEA revealed functional and quality differences between media not detected by mouse assays, supporting BEA to strengthen safety assessment while reducing animal sacrifice.
Spermatogenesis is precisely regulated by an intricate genetic network, but the biological roles of numerous testis-enriched genes remain unelucidated. This study aimed to systematically investigate the expression pattern, subcellular localization and functional significance of Testis-Expressed Gene 29 (Tex29) in mice and male fertility. Integrated molecular, cellular, and animal model approaches were employed. Tex29 mRNA and protein expression were analyzed by molecular and immunofluorescence staining techniques. CRISPR/Cas9-mediated genome editing was used to generate Tex29-knockout (KO) mice. Fertility assessment, histological examination of testes, sperm quality analysis, and transmission/scanning electron microscopy were performed on Tex29-KO mice. Additionally, whole-exome sequencing was conducted in 165 infertile men to identify Tex29 variants. Tex29 mRNA was specifically expressed in testes, first detectable on postnatal day 18 and gradually upregulated during testicular maturation. TEX29 protein was specifically localized to the acrosome of spermatids and mature sperm throughout spermiogenesis. Tex29-KO males exhibited normal fertility with litter sizes comparable to wild-type (WT) controls, and their seminiferous tubules retained intact structure with all spermatogenic stages. No significant differences in sperm concentration, viability, or motility were observed between Tex29-KO and WT mice. Although Tex29-KO sperm maintained normal overall morphology and canonical "9 + 2" axonemal structure in the flagellum, a subset showed acrosomal membrane abnormalities in the apical region. In vitro fertilization (IVF) rates and blastocyst development were uncompromised in Tex29-KO mice. Two synonymous TEX29 variants (c.66C > T, p.Asp22Asp; c.207C > A, p.Ile69Ile) were identified in 7 of 165 infertile men, and four couples carrying these variants achieved live births. TEX29 is a novel testis-specific acrosomal marker protein essential for maintaining normal acrosomal membrane integrity during murine spermiogenesis. Notably, Tex29 is dispensable for spermatogenesis and male fertility in mice. The functional role of TEX29 in human spermatogenesis and fertility remains to be fully determined due to limited clinical evidence. These findings provide valuable insights for basic research on acrosome biogenesis and male infertility associated with acrosomal abnormalities.
To assess the clinical applicability of shortened (SW) and ultra-shortened (USW) warming protocols for oocytes cryopreserved using standard vitrification (STD-V). This prospective sibling study included 436 mature human oocytes from 47 patients, all vitrified using a single standard vitrification (STD-V) protocol. Experiment 1 compared the standard warming (STD-W) with the SW protocols (2 min), and Experiment 2 compared the SW protocol with USW protocol (1 min). Oocyte survival rates were assessed 1.5 h post-warming, and oocytes were immediately fixed. The analysis of meiotic spindle organization and chromosome alignment was assessed by detailed examination of individual optical sections and corresponding full Z-stack projections encompassing the entire spindle. Mitochondrial mass was quantified from Z-stack-based projections integrating fluorescence across all optical sections. We also performed a pooled analysis to directly compare STD-W, SW, and USW groups. Survival rates were comparable across protocols. Both SW and USW protocols were associated with significantly lower odds of normal chromosomal distribution, spindle organization, and normal MII apparatus compared with the STD-W protocol, with no significant differences between SW and USW. Spindle length and pole width were increased in SW and USW versus STD-W. Mitochondrial mass was significantly reduced in SW compared with STD-W. Although survival rates were comparable, our data indicate that SW and USW protocols are associated with a higher incidence of spindle and chromosomal abnormalities than the traditional approach. This may affect oocyte developmental competence, highlighting the need for further investigation regarding the clinical use of SW/USW protocols for STD-V-cryopreserved oocytes.