Objective To elucidate the pathogenic mechanism of nephronophthisis type 12 (NPHP12) caused by compound heterozygous mutations in the TTC21B gene and to implement preimplantation genetic testing (PGT) for clinical prevention. Methods We retrospectively analyzed the clinical data of a pediatric proband with NPHP12. The impact of the identified TTC21B mutations (c.895T>C and c.1552T>C) on pre-mRNA splicing, protein structure, and stability was assessed using bioinformatics tools. Functional validation was performed through in vitro rescue experiments in renal podocytes, quantifying effects on cellular morphology, ciliogenesis, and ciliary length. For preimplantation genetic testing (PGT), SNP haplotype analysis was employed for embryo selection. Results The proband presented with renal failure and compound heterozygous TTC21B mutations (paternal c.895T>C [p.C299R] and maternal c.1552T>C [p.C518R]). Bioinformatics suggested potential splicing impacts, but minigene assays did not validate this. While c.895T>C had a minor impact on protein structure, c.1552T>C significantly altered the tertiary structure and stability. TTC21B knockdown disrupted podocyte morphology, which was fully rescued by wild-type TTC21B but only partially by either mutant. Both mutations impaired ciliogenesis and shortened ciliary length. PGT-M identified a mutation-free embryo (Embryo 1) for transfer, resulting in a healthy live birth. Conclusion The compound heterozygous mutations c.895T>C (p.C299R) and c.1552T>C (p.C518R) in TTC21B contribute to NPHP12 by disrupting ciliogenesis and podocyte morphology. These findings provide targets for PGT-based embryo selection, successfully preventing the familial recurrence of this disease.

This study aimed to investigate whether preimplantation genetic testing for aneuploidy (PGT-A) improves pregnancy outcomes in in vitro fertilization-embryo transfer (IVF-ET) cycles in patients with recurrent spontaneous abortion (RSA). A total of 216 young patients with RSA (aged <38 years) who underwent IVF were included in this study. Of these, 114 patients opted for PGT-A, comprising 64 patients with a history of embryonic chromosome abnormality (ECA) (Group A) and 50 patients without ECA (Group B). The remaining 102 patients who did not undergo PGT-A served as the Control group, including 51 patients with a history of ECA (Control A) and 51 patients without ECA (Control B). Clinical outcomes were compared between the groups. The live birth rate in Group A was significantly higher compared to Control A (P <.05), whereas no significant difference was observed between groups B and Control B (P >.05). PGT-A significantly improved clinical outcomes in RSA patients with a history of ECA, but did not show a similar benefit in those without ECA. However, owing to the retrospective nature of the analysis and potential for unmeasured confounding factors, the value of PGT-A in women with RSA warrants further investigation.

Prediction of euploidy and probability of obtaining target number of euploid embryos through preimplantation genetic testing for aneuploidy.

To quantitatively evaluate the association between sperm DNA fragmentation (SDF) and embryo euploidy rates in assisted reproductive technology (ART) cycles through a systematic review and meta-analysis. Following the PRISMA 2020 guidelines, a comprehensive search of PubMed, Web of Science, Embase, Scopus, and the Cochrane Library was conducted from inception to August 5, 2025. Eligible studies included infertile couples undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), in which SDF was assessed using validated assays and embryo euploidy was determined via preimplantation genetic testing for aneuploidy (PGT-A). The Newcastle-Ottawa Scale was used for quality assessment. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using random- or fixed-effects models based on heterogeneity. Six studies involving 1,516 ART cycles met the inclusion criteria. All studies measured SDF using the sperm chromatin structure assay (SCSA), with cutoff values ranging from 15 to 30%. Embryo chromosomal status was evaluated at the blastocyst stage using PGT-A platforms, such as next-generation sequencing (NGS), array comparative genomic hybridization (aCGH), or single nucleotide polymorphism (SNP) arrays, with whole genome amplification (WGA) applied as a pre-analytical step rather than a detection method. Meta-analysis revealed no significant association between high SDF and embryo euploidy when using the 15% cutoff (pooled OR = 0.897; 95% CI 0.741-1.085; I2 = 0.0%). At the 30% cutoff, high SDF (DFI ≥ 30%) was associated with lower embryo euploidy rates (pooled OR = 0.742; 95% CI 0.558-0.988; I2 = 62.2%). Elevated SDF, particularly above 30%, is associated with a reduced likelihood of obtaining euploid embryos in ART cycles, suggesting a potential threshold-dependent effect of sperm DNA integrity on embryo chromosomal normality. These findings support the integration of SDF assessment into the evaluation of selected couples, especially in cases of recurrent ART failure or advanced maternal age. Further prospective studies with standardized SDF protocols and uniform PGT-A methods are warranted to validate these results.

Intracytoplasmic sperm injection (ICSI) was first developed to overcome male factor infertility. ICSI has increased in uptake globally, including in cases where its use is non-essential for fertilisation. To identify temporal trends in the use of, and indications for ICSI in an Australian context. A statewide descriptive cohort study examining the trends in ICSI uptake and reported indication/s for ICSI use. The cohort included women undergoing IVF between 2005 and 2017 at IVF clinics across Victoria, Australia that resulted in a birth after 20 weeks' gestation. The dataset comprised 32 102 assisted reproduction cycles: 22 873 (71.3%) ICSI and 9229 (28.7%) conventional IVF. In 2005, ICSI accounted for 60.6% (1182/1952) of cycles, increasing to 79.5% (2344/2947) by 2017 (ptrend < 0.001). Testicular sperm retrieval as an indication for ICSI remained consistent over time (ptrend = 0.15). Male factor infertility as an indication decreased over time (ptrend = 0.007). Vitrified oocyte thaw (ptrend = 0.016) and 'unexplained subfertility' (ptrend = 0.30) cycles did not surpass 1.7% (39/2293) and 0.4% (9/2048), respectively of total cycles in any year. Donor sperm (ptrend = 0.001), pre-implantation genetic testing (ptrend = 0.004), female factors associated with poor IVF outcome (ptrend = 0.005) and advanced maternal age (ptrend = 0.005) all increased as indications for ICSI over time. 'Unspecified' indication accounted for the majority of ICSI cycles after 2008 (ptrend = 0.015). During our study period, the total use of ICSI increased by 18.9%. Notably, most of these cycles were not medically indicated.

Exploring experiences and perceptions of fertility and reproductive technology among people with sickle cell disease

Reframing analytical validation in preimplantation genetic testing: phase 1.

Ongoing surveillance of analytical platforms in preimplantation genetic testing: phase 4.

Clinical validation of embryo prognostic tests; determining predictive value: phase 2.

Procedural safety and ongoing monitoring and evaluation of embryo diagnostics.

De novo inherited Xq25 deletion: hints from preimplantation genetic testing in alobar holoprosencephaly.

From innovation to practice: evaluating the evidence for preimplantation genetic testing for aneuploidy.

Can a prediction model classify IVF patients into distinct prognostic groups based on their expected yield of euploid blastocysts? Five distinct prognostic groups were identified, with chance of obtaining at least one euploid blastocyst ranging from <1% to 2% in very poor to ∼95% in very good prognosis groups. Euploid blastocyst yield is a critical determinant of IVF success. While female age strongly influences embryo euploidy, other factors like ovarian reserve markers, partner age, and BMI may also contribute. Current approaches rely on basic and somewhat arbitrary classification of ovarian reserve markers and patient age, which are unable to represent the granular multidimensional relationship between them. A systematic approach to classify patients based on their expected euploid yield would enable better treatment planning and patient counseling. A retrospective analysis of 10774 IVF cycles from 8256 couples undergoing pre-implantation genetic testing for aneuploidy (PGT-A) of all available blastocysts from years 2020 to 2023 and a temporal validation cohort of 2089 cycles of 2089 patients from year 2024. The prediction model was developed using generalized additive models for location, scale, and shape using a development cohort from 2020 to 2023, and cross-validated through exhaustive 5-fold cross-validation. Temporal validation was performed using an entirely separate cohort from 2024. Model performance was assessed through calibration plots and discrimination metrics. Couples undergoing IVF with PGT-A of all their available blastocysts were included. Model included female age, partner age, anti-Müllerian hormone (AMH), antral follicle count, and BMI as predictors. Non-linear associations were captured using neural networks and restricted cubic splines. Missing data were handled using multivariate imputation by chained equations. The median female age was 36.3 years (IQR: 33.3-39.5) and AMH was 2.0 ng/ml (IQR: 1.0-3.8). Models for predicting ≥1, ≥2, and ≥3 euploid blastocysts yield achieved very good discrimination performance in 5-fold cross-validation samples with mean AUCs of 0.834, 0.849, and 0.861, respectively. Models showed negligible shrinkage (<1%) between training and cross-validation sets with near-perfect calibration slopes (mean: 1.00, IQR: 0.99-1.01) and intercepts (mean: 0.015, IQR: 0.00-0.03). Using predicted absolute counts of euploid blastocysts, five distinct prognostic groups were created based on predicted euploid blastocyst yield. Patients in the very poor prognosis group had 98.3% probability of obtaining no euploid blastocysts after stimulation while the probabilities were 80.2%, 47.5%, 15.8%, and 4.7% in poor, borderline, good, and very good prognosis groups. The chances of obtaining ≥3 euploid blastocysts were 79.8%, 43.7%, 8.9%, 0.2%, and 0% in very good, good, borderline, poor, and very poor prognosis groups. In the temporal validation set (n = 2089), which constituted the first cycles of patients that were treated, the rates of no euploid blastocysts obtained at the end of stimulation were 100.0%, 82.6%, 47.4%, 13.1%, and 4.4% in the very poor, poor, borderline, good, and very good prognosis groups. The rates of three or more euploid blastocysts obtained at the end of stimulation in the temporal validation set were 83.5%, 51.0%, 10.7%, 0.6%, and 0.0% for very good, good, borderline, poor, and very poor prognosis groups. The FORTUNE (Forecasting Outcomes of Reproductive Treatments Using artificial Networks) model is available for use and further validation at https://epsilonkappa-analytics.shinyapps.io/FORTUNE and App Store for iOS mobile devices https://apps.apple.com/en/app/fortune-ivf/id6747190429. Single-center study design may limit generalizability. The model does not account for laboratory-specific factors or stimulation protocols. This novel classification system provides objective, personalized counseling for IVF patients regarding expected euploid yield, enabling better-informed decision-making about treatment options and number of planned stimulation cycles. There was no funding needed for this study. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. N/A.

The goal of embryo diagnostics and prognostics testing.

Use of preimplantation genetic testing for polygenic disorders (PGT-P): an Ethics Committee opinion.

Infertility affects a significant portion of couples globally. In vitro fertilization (IVF) is a widely utilized assisted reproductive technology (ART). Despite significant advancements, IVF success rates remain suboptimal, which necessitates innovative approaches to improve embryo selection and predict the outcomes. Traditional methods, including morphological grading and preimplantation genetic testing (PGT), possess limitations, necessitating the development of innovative, non-invasive biomarkers. This narrative review sought to investigate novel biomarkers, such as oxidativestress (OS) indicators like malondialdehyde and advanced oxidation protein products, antioxidants including glutathioneperoxidase and catalase, cell-free DNA (cfDNA), immunologic markers, and microRNAs (miRNAs) extracted from follicularfluid (FF), blood, and endometrial tissue, which may serve as predictors of ART outcomes. A narrative review ofpublished studies investigating non-invasive biomarkers in human ART was conducted. Literature was sourced from databasesincluding PubMed, Scopus, and Web of Science, focusing on studies reporting oxidative stress markers, antioxidant enzymes,miRNAs, cfDNA, and immunologic markers in FF, blood, or endometrial samples. Elevated OS markerswere linked to low-quality embryos, while high levels of antioxidants were linked to higher quality of oocytes and embryos.Moreover, certain miRNAs in FF exhibited potential as indicators of embryo viability and implantation success, potentiallyenabling more precise and personalized IVF approaches. Incorporating these emerging biomarkers into standardclinical practice can improve prediction methods. Future research should focus on confirming these biomarkers in variouspopulations and creating standardized protocols to promote their integration into reproductive medicine which can lead to betterfertility treatments and help couples deal with the emotional and psychological problems caused by infertility.

Transcriptomics has emerged as a crucial methodology for deciphering gene expression and regulatory networks, offering novel insights and robust scientific support for advancing assisted reproductive technologies (ART). This review synthesizes current knowledge on the transcriptome profiles of germ cells and early embryos, elucidating key molecular signatures associated with developmental potential, maturation processes, and embryo quality. It further demonstrates the application of transcriptomics in preimplantation genetic testing (PGT), enhancing embryo selection precision through the screening of chromosomal abnormalities and genetic disorders. Additionally, the review highlights the significant value of transcriptomic approaches in evaluating embryonic developmental potential and optimizing in vitro culture conditions. Collectively, these applications provide a strong foundation for improving ART success rates and advancing personalized treatment strategies.

To propose an improved blastocyst grading system by integrating dynamic time-lapse parameters with conventional morphological criteria, aiming to enhance embryo selection during assisted reproduction. This retrospective cohort study evaluated 1,182 embryos derived from 433 cycles of intracytoplasmic sperm injection. Time-lapse parameters included morula compaction status, blastocyst collapse frequency, and fragmentation of the inner cell mass and trophectoderm. These findings were analyzed in conjunction with standard morphological grading and embryo scoring systems based on implantation data. Embryo outcomes were stratified by chromosomal status determined through preimplantation genetic testing for aneuploidy. Fully compacted morulae were associated with higher morphological grades; however, the proportion of chromosomally normal embryos did not differ significantly when compared to those derived from partially compacted morulae, suggesting that static morphology alone may not accurately reflect developmental potential. Although most blastocyst collapse events were minor, they correlated with increased fragmentation and reduced embryo quality. Fragmentation of the inner cell mass and/or the trophectoderm was associated with significantly lower rates of chromosomal normalcy. Logistic regression analysis confirmed fragmentation as the strongest predictor of aneuploidy, outperforming both conventional grading systems. This integrated evaluation model, combining static morphology with dynamic developmental events, offers a more comprehensive and accurate approach to assessing embryo viability. By identifying embryos with higher risk of chromosomal abnormalities, this strategy may enhance embryo selection, improve reproductive outcomes, and contribute to more individualized care in assisted reproduction.

Existing methods to image chromosome segregation errors are not suitable for studying human embryos at advanced preimplantation stages. As chromosomal errors are a leading cause of miscarriage and infertility, it remains unclear whether missegregation arises postfertilization. Here we optimize nuclear DNA labeling via messenger RNA electroporation and apply light-sheet live imaging to reveal chromosome segregation errors immediately before implantation. We show that embryos at advanced preimplantation stages display missegregation, including multipolar spindle formation, lagging chromosomes, misalignment and mitotic slippage. Most lagging chromosomes are passively inherited rather than reincorporated. To trace individual nuclei, we developed an open-source, semi-automated segmentation method using a customized deep learning model optimized for variability in embryo size, shape and signal. With this approach, we find most labeled cells remain externally positioned, consistent with placental rather than inner cell mass fate. Our findings raise questions about clinical uses of preimplantation genetic testing for aneuploidy, while providing broadly applicable imaging and segmentation methods for studying diverse cellular structures in human embryos.

Embryo selection and preimplantation genetic testing (PGT) are used in clinical practice with the aim of improving pregnancy outcomes in patients. However, it remains uncertain whether different PGT techniques improve the live birth rate in patients compared to conventional in vitro fertilization (IVF). In this single-center retrospective cohort study, we included couples undergoing their first IVF-assisted conception cycle with only their first single frozen embryo transfer. All couples were fertilized by intracytoplasmic sperm injection (ICSI) and had three or more high-quality blastocysts on the fifth day of embryo culture. The PGT group screened the optimal blastocysts for transfer by their counterpart technique, while the conventional IVF group selected blastocysts based on morphological criteria. Clinicians select different controlled ovarian hyperstimulation protocols and endometrial preparation protocols based on the patient's individual circumstances. The primary outcome was to observe the live birth rate of first frozen single-embryo transfer in each group. A total of 409 couples were enrolled in the study, of which 219 were in the PGT group, 87 were PGT-A, 22 were PGT-M, 110 were PGT-SR, and 190 were in the IVF group. The average age of patients in the PGT group was 32.90years, while 31.53years in the IVF group. The live birth rate was higher in the PGT group (46.12%) compared to that of the conventional IVF group (34.74%) (p = 0.0195, OR 0.6218, 95% CI 0.4161-0.9189). The live birth rate in the PGT-A subgroup (48.28%) was also higher than that in the IVF group (p = 0.032, OR 0.5703, 95% CI 0.3380-0.9610). Different controlled ovarian hyperstimulation protocols and endometrial preparation protocols had no effect on the live birth rate of PGT. For first frozen single-embryo transfer, PGT, particularly PGT-A, can improve live birth rates compared to conventional IVF. Different controlled ovarian hyperstimulation protocols and endometrial preparation protocols had no effect on the live birth rate of PGT.