This study aimed to evaluate the effectiveness of changing the insemination method in women who experienced poor embryonic development during the preceding cycle. A total of 15,886 conventional IVF in 9,311 women, performed between August 2015 and June 2023, were included in this study. Of these, 270 couples experienced IVF failure due to poor embryonic development in the first oocyte retrieval (OR) cycle, which was cancelled before transfer. The patients were stratified based on whether or not they switched to ICSI for subsequent attempts. Cumulative live birth rates (CLBRs) and a series of secondary outcomes were compared. The embryo utilization, high-quality embryo, blastocyst formation, implantation, cumulative clinical pregnancy, CLBR and miscarriage rates were comparable between the two groups, whereas the fertilization rate per oocyte retrieved was significantly lower in the ICSI group during the second OR cycle (60.76% vs. 70.42%, p < 0.001) and all OR cycles (60.02% vs. 71.69%, p < 0.001). Furthermore, the CLBRs in the ICSI and IVF groups after up to seven OR cycles were 41.35% and 36.84%, respectively. Most patients achieved live births during the second OR cycle (58.33%, ICSI vs. 62.86%, IVF). ICSI did not improve clinical or embryonic outcomes in women who experienced poor embryonic development in their preceding cycle.

Infertility is a primary health issue affecting about 15% of couples of reproductive ages worldwide, leading to physical, mental, and social challenges. Advances in nanobiotechnology and regenerative medicine are opening new therapeutic horizons for infertility by developing polysaccharide-based nanostructured biomaterials. This review explores the role of tissue engineering and regenerative medicine in infertility treatment, explicitly focusing on the promising potential of polysaccharide-based hydrogels. In this context, using these biomaterials offers unique advantages, including biodegradability, biocompatibility, and the ability to mimic the natural endometrial microenvironment, making them highly effective for applications in endometrial regeneration, ovarian tissue engineering, spermatogenesis support, and controlled drug delivery. This review discusses the various properties and uses of polysaccharide-based hydrogels, like alginate, hyaluronic acid, and chitosan, in helping to restore reproductive function. While these materials hold great promise, some notable challenges to their clinical use include issues like rapid degradation, mechanical instability, and potential immune reactions. Future research should focus on developing hybrid hydrogels, investigating advanced fabrication techniques, and testing these materials in clinical settings. By combining findings from recent studies, this review aims to provide a solid foundation for researchers and clinicians looking to discover new and effective strategies for treating infertility, ultimately connecting research efforts with practical applications in healthcare.

To evaluate the predictive ability of ooplasmic fluidity assessment during piezo-intracytoplasmic sperm injection (ICSI) for unintentional membrane rupture (UMR) and oocyte degeneration.We established an optical-flow analysis method that detects ooplasm movement in piezo-ICSI videos. Videos of 104 oocytes recorded during piezo-ICSI at our institute between January 2020 and January 2024 were retrospectively analyzed.To evaluate the apparent velocities in the ooplasmic region, we analyzed the vertical velocity component of the obtained optical flow between the start of puncture and the rupture of the oolemma (1st period) and between sperm injection and pipette withdrawal (2nd period). The velocity distributions between groups were compared using the 95th, 99th, and 99.5th percentiles. The 95th, 99th, and 99.5th percentiles of the survival group (n = 76) were significantly higher than those of the degeneration group (n = 28) during the 1st period (P < 0.001). Similarly, the 95th, 99th, and 99.5th percentiles of the survival group were significantly higher than those of the degenerative group during the 2nd period (P < 0.05). The 95th, 99th, and 99.5th percentiles without UMR (n = 52) were significantly higher than those with UMR (n = 50) during the 1st period (P < 0.001). Similarly, the 95th, 99th, and 99.5th percentiles without UMR were significantly higher than those with UMR during the 2nd period (P < 0.001). Greater ooplasmic fluidity was correlated with lower incidence of UMR and oocyte degeneration. Elucidation of the mechanisms underlying this phenomenon may help reduce UMR and oocyte degeneration.

Many national recommendations have been made regarding practice patterns of assisted reproductive technology utilization (intracytoplasmic sperm injection (ICSI), preimplantation genetic testing (PGT), and blastocyst stage embryo transfer). This study seeks to assess individual clinic changes in practice behaviors from 2014 to 2020 in response to national guidelines.

This retrospective cohort study examined data from individual clinics using the Society of Assisted Reproductive Technology Clinical Outcome Reporting System (SART-CORS) database. Each clinic was weighed individually and equally regardless of clinic volume to avoid skewing national data with larger-volume clinics. The trends of ICSI, PGT, and blastocyst stage embryo transfer were examined across five age groups (< 35, 35-37, 38-40, 40-42, and > 42).

ICSI usage from 2014 to 2020 increased in all age groups (p < 0.001). In 2020, more than half the clinics in this study utilized ICSI in ≥ 90% of IVF cycles (per oocyte retrieval). Utilization of prolonged embryo culture increased in all age groups across the seven years of this study, with a more extensive adoption of this practice in patients ≤ 40 years (p < 0.001). Between 2014 and 2020, PGT usage increased by 3-4 times across all age groups (p < 0.001). By 2020, PGT was performed after egg retrievals in approximately 40 to 50% (mean) of patients < 42 years old.

Nationally, the application of all three reproductive technologies (ICSI, PGT, and blastocyst stage transfer) has drastically increased. The authors suspect that these technology expansions are interrelated and may be associated with the rise of rates of PGT.

Objectives: To compare surgical outcomes and sperm retrieval rates (SRRs) between conventional microsurgical-assisted testicular sperm extraction (m-TeSE-Group A) and testicular sperm extraction performed with surgical loupes (l-TeSE-Group B) in adult males with non-obstructive azoospermia (NOA). Methods: A multicentric prospective randomized trial (ethics committee no. 202/2022) in accordance with the CONSORT guidelines was conducted from March 2022 to April 2024. Adult males with NOA without genetic alterations who signed the informed consent were enrolled. SRRs, intra- and postoperative complications (according to the Clavien-Dindo classification), and hormonal profile changes were considered as outcomes during the follow-up period. Results: A total of 42 NOA patients were enrolled. The median age was 35 years (IQR: 33-49). The preoperative median FSH was 16.5 mIU/mL (IQR: 11.6-22.5) and the total testosterone was 4.6 (3.5-5.6). Overall, the SRR was 22.6%, with sperm retrieved from 19 testes. Histopathological findings reported Sertoli cell-only syndrome (SCOS) in 46.4% (39 cases), hypospermatogenesis in 26.2%, and germ cell arrest in 26.2% of the patients. No intraoperative complications were recorded. The postoperative complications were minimal (Clavien-Dindo grade I), but no significant differences were recorded in-between the two surgical approaches. Considering the operative time of the testicular exploration alone, Group B seemed to be faster than the m-TeSE, with a median time saving of 8 min (p < 0.01). Conclusion: The use of surgical loupes was safe and comparable with m-TeSE in terms of the SRRs and complication rates. L-TeSE offered a reduction in the operative time compared with m-TeSE.

Does a short co-incubation of gametes in conventional IVF affect post-insemination outcomes and embryo morphokinetics?

Sibling oocyte randomized pilot study conducted between December 2020 and March 2023. Eligible couples (n = 55) were women aged 18-43 years with BMI 35 km/m2 or lower and male normal semen parameters. Cumulus oocyte complexes (COC) (six to 12) were randomized in a 1:1:2 proportion in long (16-18 h) or short (2 h) co-incubation IVF exposure and ICSI, respectively. All oocytes inseminated via IVF were monitored by time-lapse. Blastocysts graded BL3CC or higher underwent trophectoderm biopsy on days 5, 6 or 7, and were analysed by next-generation sequencing.

A total of 954 COC were distributed to long co-incubation (n = 235 [24.6%]), short co-incubation (n = 235 [24.6%]) and ICSI (n = 484 [50.7%]). In total, 202 were MII after long co-incubation, 209 after short co-incubation and 394 at ICSI. Regression analyses showed that short co-incubation groups (OR 0.72, 95% CI 0.46 to 1.11, P = 0.139) and ICSI (OR 1.43, 95% CI 0.95 to 2.15, P = 0.090) did not significantly affect normal fertilization rates (2PN) compared with long co-incubation. Usable blastocysts per MII, per 2PN, and euploid blastocysts per MII, per 2PN and per tested blastocysts were similar among the groups. The rate of embryo arrest (hazard ratio [HR] 0.79, 95% CI 0.57 to 1.11, P = 0.183) and timings to reach morphokinetic milestones among embryos reaching blastulation were similar between IVF groups (P > 0.05 for all).

A short 2-h co-incubation of gametes in IVF yields comparable fertilization, blastocyst and euploidy rates without adverse embryo morphokinetic events, compared with standard overnight co-incubation.

This manuscript explores the intricacies and nuances of the Intracytoplasmic Sperm Injection (ICSI) procedure, drawing on insights from three decades of experience at a specialized center managing numerous cases of male factor infertility. Our center is comprised of an embryology laboratory, an andrology and assisted fertilization laboratory, and a Preimplantation Genetic Testing for Aneuploidy (PGT-A) laboratory, each fostering specialized expertise independently. Collaboration among these laboratories, alongside reproductive physicians and urologists, ensures comprehensive feedback and optimal care for patients undergoing infertility treatment. The manuscript specifically focuses on the andrology laboratory's pivotal role in evaluating and treating infertile patients, highlighting critical preparations for the ICSI procedure, and the key considerations essential to its successful implementation, including the selection of the ideal spermatozoon, oocyte dysmaturity, proper equipment, and most importantly the execution of the procedure itself.

To investigate whether idiopathic nonobstructive azoospermia (iNOA) has its own transcriptomic signature.

Testicular tissue biopsies were retrieved, processed, and prepared for ribonucleic acid (RNA) extraction from 26 consented patients diagnosed with iNOA. Samples were grouped into four pools based on the presence of testicular spermatozoa: two replicate pools for "No presence" (Null-spz-1 and Null-spz-2 pools), one for "High presence" (High-spz pool), and one for "Rare presence" (Rare-spz pool). A second set of replicate pools (CF-1 and CF-2) were used from patients with obstructive azoospermia (OA) and served as controls. RNA sequencing (RNA-seq) and comparative transcriptomics analysis were performed, followed by differential gene expression analysis focused on protein-coding genes only. Differentially expressed genes (DEGs) exclusively upregulated or downregulated were further analyzed using the Gene Ontology (GO), STRING, and Kyoto Encyclopedia of Genes and Genome bioinformatic platforms.

Males in whom iNOA was diagnosed.

Testicular biopsies from men in whom iNOA was diagnosed.

Protein-coding DEGs.

A significantly altered transcriptomic profile of protein-coding genes was identified in the testicular tissues from men with iNOA. A total of 3,858 genes exhibited dysregulated expression, with 1,994 genes being exclusively downregulated and 1,734 upregulated. Biological processes such as male gamete generation (GO:0048232) and meiotic cycle (GO:0051321) were significantly enriched by the downregulated DEGs whereas the upregulated DEGs enriched BPs such as regulation of cell death (GO:0010941), regulation of cell adhesion (GO:0030155), and defense response (GO:0006952). Interactome analysis identified hub genes among the downregulated DEGs, including PCNA, PLK1, MCM4, CDK1, CCNB1, AURKA, CCNA2, and CDC6, and among the upregulated DEGs, including EGFR, RELA, CTNNB1, MYC, JUN, SMAD3, STAT3 NFKB1, TGFB1, and ACTB. In addition, Kyoto Encyclopedia of Genes and Genome analysis demonstrated that pathways such as cell cycle (hsa04110) and oocyte meiosis (hsa04114) are primarily affected by the downregulated genes, whereas the upregulated genes mainly affected pathways such as the focal adhesion (hsa04510) and the PI3-Akt signaling pathway (hsa04151).

A distinct messenger RNA expression profile and altered transcriptomic activity were identified in the testicular tissues of men with iNOA.

University of Thessaly 1, 15.04.2016 and the Greek National Authority 701/15.9.2017.

This study aimed to compare sperm DNA fragmentation (SDF) levels between ejaculate and testicular sperm and evaluate clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles using testicular sperm (T-ICSI) versus ejaculate sperm (E-ICSI) in males with high ejaculate SDF, prior ICSI failures, or severe male infertility. A systematic review of major databases and a subsequent meta-analysis were performed to compare clinical outcomes in men with high SDF, oligozoospermia, or prior ICSI failures undergoing T-ICSI or E-ICSI. Thirteen studies met the inclusion criteria. Outcomes analyzed included SDF levels, fertilization rate (FR), clinical pregnancy rate (CPR), live birth rate (LBR) per embryo transfer (ET), and miscarriage rate (MR) per pregnancy. The mean difference (MD) and odds ratio (OR) were calculated for each outcome. Paired assessments of SDF showed significantly lower levels in testicular sperm compared to ejaculated sperm (MD = -25.42 [-31.47, -17.30], p < 0.00001). While no significant difference in FR was observed in T-ICSI cycles overall (OR = 0.94 [0.74, 1.20]), a subgroup analysis revealed significantly higher FR with E-ICSI in men with oligozoospermia and no prior ICSI failures (OR = 0.61 [0.52, 0.71], p < 0.00001). CPR was significantly higher in T-ICSI cycles (OR = 2.13 [1.35, 3.36], p < 0.001; n = 540 ET), along with a significantly lower MR (OR = 0.31 [0.14, 0.70], p = 0.004; n = 35) and increased LBR (OR = 2.40 [1.32, 4.36], p = 0.004; n = 446 ET). In conclusion, using testicular sperm in cases of elevated ejaculate SDF, oligozoospermia, or prior failed ICSI cycles enhances the selection of sperm with lower DNA damage, leading to improved pregnancy rates, reduced miscarriage rates, and higher live birth rates. However, the studies included were rated as having a moderate to serious risk of bias. Further well-designed randomized controlled trials are necessary to confirm these findings with stronger evidence.

Intracytoplasmic sperm injection (ICSI) is an assisted reproductive technique (ART) mainly used to overcome severe male factor infertility problems in humans and animals. However, in cattle, one of the most demanded species for its meat and milk, the efficiency of this technique is low. The present study compared the effect of the piezoelectric and conventional injection systems on the preimplantational development and quality of bovine embryos generated by ICSI. Evaluations of the conditions for performing the Piezo-ICSI procedure showed that the application of a strong pulse (I4S7) was more effective in damaging the sperm plasma and acrosomal membranes prior to injection, compared to a soft pulse (I2S2, P < 0.05). In addition, Piezo-ICSI embryos without the application of exogenous activators achieved similar levels of development as Piezo-ICSI embryos activated with ionomycin and anisomycin (P > 0.05). When comparing conventional and piezoelectric injection systems, no significant differences in embryo development were observed (P > 0.05). However, embryos generated by Piezo-ICSI showed a higher embryo quality in terms of total cell number (P < 0.05). In addition, Piezo-ICSI embryos showed an expression profile of genes essential for embryonic development similar to IVF embryos (P > 0.05), in contrast to conventional ICSI-derived embryos, which presented overexpression of CASP3 and IFNT2 (P < 0.05). In conclusion, we confirmed that Piezo-ICSI is a more convenient approach than traditional ICSI, since does not require exogenous activation and generate embryos of better quality, regarding the total number of blastomeres and the pattern of gene expression observed.

Microdissection TESE has been considered the "gold standard" for retrieving testicular sperm in cases of non-obstructive azoospermia (NOA) despite limited scientific support. Here we compare all aspects of microdissection TESE with testis fine needle aspiration mapping (FNA Mapping) and directed TESE procedures for men with NOA.

We examine the history of testicular sperm extraction techniques and the rise of advanced technologies with a focus on microdissection TESE and FNA mapping. We summarize the published literature regarding the success rates, complications, and limitations of these two methods.

As there are no randomized controlled trials, the best data come from the Cochrane Reviews, which include meta-analyses concluding that the simplest and safest methods of sperm retrieval should be chosen. Although microdissection TESE is popular, recent reports have questioned its value due to the significant hypogonadal consequences. Among alternative procedures, FNA Mapping is a viable and less invasive alternative to microdissection TESE in finding testicular sperm in NOA patients.

Alternatives to microdissection TESE procedures such as FNA Mapping offer several advantages that include similar sperm retrieval success rates, but also less invasiveness and improved understanding of the pathophysiology of NOA.

To evaluate whether the ZyMōt™ Multi 850 μl sperm separation device (SSD) effectively recovers motile spermatozoa from cryopreserved ejaculates and compare its effect on key embryology outcomes including fertilization, cleavage stage, and total and top-quality blastocyst formation rates to the traditional Density Gradient Centrifugation (DGC) method.

In this prospective, single-center, controlled study, we used fresh sibling donor oocytes and non-donor cryopreserved ejaculates. In total, 150 couples participated in this study. At least eight MII donor oocytes were allocated to each couple split into two arms. One arm underwent ICSI with the control DGC-processed sample, and the other arm processed with SSD.

No significant difference on fertilization and cleavage stage embryo rates was observed between the two techniques. We observed a significant increase in the percentage of total (SSD: 74.03 ± 23.47% vs. DGC: 67.86 ± 23.92%; p = 0.016) and top-quality (SSD: 66.38 ± 24.94% vs. DGC: 60.98 ± 24.40%; p = 0.035) blastocysts formed post-SSD processing. Sub-analysis showed that this increase remained significant for the WHO-normal group (n = 118), but not for the WHO-abnormal group (n = 32).

The SSD was successfully applied in all 150 cases, providing adequate numbers of spermatozoa to undergo ICSI. Additionally, SSD significantly improved blastocyst development rates; however, this was of limited clinical impact considering the minor improvement on the average number of top-quality blastocysts. It can be hypothesized that this positive contribution may be stronger and clinically significant when a larger number of oocytes is used or in homologous oocyte ICSI cycles, where the repair mechanisms of the oocytes may insufficient for promoting healthy embryo development.

Today about 12.5% of all children are conceived using assisted reproductive technology. Intracytoplasmic sperm injection (ICSI) was introduced without knowledge of the possible long-term adverse consequences on the offspring health. We aimed to examine risk of neurodevelopmental disorders, asthma, epilepsy, inflammatory bowel disease, rheumatoid arthritis, thyroid disease, and diabetes in the offspring conceived with ICSI, compared with offspring conceived with in vitro fertilization (IVF) and natural conception.

All offspring conceived using ICSI, IVF, and natural conception in Denmark from July 9th, 1994 until December 31st, 2019 were included in this nationwide cohort study using Danish national health registries.

Median follow-up time was 10 years (6-16 years, interquartile range (IQR)) for offspring conceived with ICSI (n = 22,906), 13 years (7-20 years, IQR) for offspring conceived with IVF (n = 32,458), and 13 years (8-20, IQR) for offspring born after natural conception (n = 1,397,791). Several of the outcomes occurred rarely. The most frequent outcome in offspring conceived with ICSI was neurodevelopmental disorder which did not show an increased risk when comparing offspring conceived with ICSI with offspring conceived with IVF (hazard ratio 1.02 (95 % CI: 0.92;1.12)) and natural conception (hazard ratio 0.96 (95 % CI: 0.88;1.40)).

Reassuringly, there were no statistically significantly increased risks of the examined outcomes in offspring conceived with ICSI. Before any firm conclusions are made, this study should be replicated in large populations with longer follow-up time.

Among couples with infertility and normal total sperm count and motility, can sperm morphology be used as a biomarker to identify couples who benefit more from ICSI over conventional IVF (c-IVF) on fertility outcomes?

Based on this secondary analysis of a large randomized clinical trial (RCT), sperm morphology has limited value as a biomarker to identify couples who benefit more from ICSI over c-IVF on live birth, ongoing pregnancy, clinical pregnancy or total fertilization failure.

Our recent RCT showed that ICSI did not result in higher live birth rates in couples with normal total sperm count and motility. It is unclear whether sperm morphology can be used as a biomarker to identify couples who benefit more from ICSI over c-IVF in this population.

This was a secondary analysis of an open-label, multi-centre, RCT comparing ICSI versus c-IVF in 1064 couples with infertility and normal total sperm count and motility. In this secondary study, we evaluated the effectiveness of ICSI over c-IVF in relation to sperm morphology.

Couples were eligible if they had ≤2 previous IVF/ICSI attempts, and the male partner had normal total sperm count and motility according to the fifth edition of the WHO laboratory manual for the examination and processing of human semen. Sperm morphology was measured from samples obtained during the first consultation and data for sperm morphology were available in partners of all participants in this trial. The outcomes of interest were live birth, ongoing pregnancy, clinical pregnancy, and total fertilization failure. We first conducted a logistic regression analysis with an interaction term (sperm morphology as a continuous variable by treatment (ICSI versus c-IVF)) on the four outcomes. We also used restricted cubic spline analysis to evaluate non-linear interaction and plotted the treatment effects of ICSI over c-IVF at different sperm morphology levels and the predicted probability of these outcomes in both ICSI and c-IVF groups.

The median proportion of sperm with normal morphology in both groups was 3% (Interquartile range 1-6%). Live birth rates were (184/532) 34.6% for ICSI versus (166/532) 31.2% for c-IVF. No significant interaction was found between sperm morphology and treatment effect of ICSI versus c-IVF on the rates of live birth, ongoing pregnancy, clinical pregnancy, and total fertilization failure (P = 0.181, 0.153, 0.168, and 0.788 respectively). In the analyses using restricted cubic splines, no evidence of interaction between sperm morphology and the treatment effect was found. Interaction figures showed that the treatment effect of ICSI over c-IVF at different sperm morphology levels was fluctuating around no effect line, and the predicted outcomes for the two groups were mostly overlapping at different sperm morphology levels.

This secondary analysis may be underpowered to detect a difference in treatment effects at different sperm morphology levels due to relatively small number of events at some sperm morphology levels. Moreover, sperm morphology assessment was performed during the first consultation, rather than on the day of randomization.

In couples with infertility and normal total sperm count and motility, sperm morphology has a limited role as a biomarker to identify couples who benefit more from ICSI over c-IVF on fertility outcomes.

This study was funded by My Duc Hospital, Ho Chi Minh City, Vietnam. RW was supported by an NHMRC EL Investigator Grant (GNT2009767). LNV has received speaker and conference fees from Merck, grant, speaker, conference fees from Merck Sharpe and Dohme, and speaker, conference, and scientific board fees from Ferring. TMH has received speaker fees from Merck, Merck Sharp Dohme, and Ferring. BWM reports consultancy, travel support and research funding from Merck and consultancy for Organon and Norgine. BWM holds stock from ObsEva.

NCT03428919.

The objective is to evaluate the effectiveness and safety of calcium ionophore as an artificial oocyte activation (AOA) method on pregnancy outcomes in different groups of intracytoplasmic sperm injection (ICSI) patients, providing potential evidence to establish consensus on the indications of AOA.

A systematic comprehensive search was performed in Medline, Embase, the Cochrane Library, and Google Scholar databases. Studies published from January 2014 to June 2024 were searched for analysis. All studies that compared ICSI with AOA-ICSI in routine indications composing impaired fertilization or embryo developmental arrest in previous cycles, or male-factor infertility were included.

Twelve studies were included in the meta-analysis. AOA-ICSI was associated with the increase in the overall fertilization rate (OR 1.99, 95% CI 1.16-3.41) and live birth rate (OR 4.58, 95% CI 1.52-13.80). All secondary outcomes including cleavage, blastocyst, high-quality embryo, implantation, biochemical pregnancy, clinical pregnancy presented superiority or equivalence in AOA-ICSI. And the use of calcium ionophore did not increase the miscarriage rate (OR 0.43, 95% CI 0.08-2.43). In subgroup analysis, AOA-ICSI exhibited a more significant effect on patients with indications of no or low fertilization. However, in patients with non-fertilization factors, no statistically significant improvements were observed in all outcomes.

Calcium ionophore is an effective artificial oocyte activation approach to improving pregnancy outcomes after ICSI, particularly in cases with indications of fertilization factors, providing further support for the application of AOA in specific populations. Further validation is needed to comprehensively establish the safety of AOA.

PROSPERO registration number CRD42024551481.

Aim: The current sperm selection procedure for intracytoplasmic sperm injection has limited ability to detect structural and functional abnormalities of the spermatozoa. The aim of this study was to investigate whether the degree of sperm tail swelling observed during hypo-osmotic swelling test (HOST) may predict sperm quality.Materials & methods: Sixty semen samples were collected from men investigated for couple infertility. For each sample, sperm parameters, HOST and sperm chromatin status were evaluated. The relationship between the different HOST-induced tail swelling patterns ('a' to 'g') and sperm quality was evaluated.Results: The HOST significantly correlated with higher sperm motility and vitality, and with better morphology and nuclear quality. The HOST grades 'b' and 'c' were associated with better motility (p < 0.05 and p < 0.01, respectively) and morphology (grade 'b', p < 0.001). While grade 'd' was associated with better motility and count (p < 0.05).Conclusion: Our results show that the pattern of sperm tail swelling is linked to sperm functional integrity. HOST grades 'b' and 'd' were associated with better sperm quality, suggesting their preferential use during routine sperm selection for intracytoplasmic sperm injection.

To introduce an innovative noncontact method for denudation process of cumulus-oocyte complexes (COCs) for intracytoplasmic sperm injection (ICSI).

We designed and fabricated novel acoustohydrodynamic tweezers (AHTs) to perform contactless denudation and tested them in a mouse model. Cumulus removal efficiency, preimplantation development, and live birth were assessed and compared with those in conventional manual pipetting (MP) denudation.

Fourteen female B6D2F1/J mice (approximately 4 weeks of age), nine male B6D2F1/J mice (6-12 weeks of age), and 28 CD-1 female mice (approximately 6 weeks of age) were used for experiment.

We designed a contactless platform for oocyte denudation on the basis of the principle of focalized acoustic waves. We first investigated the acoustic intensity and thermal variability by measuring the surface displacement and temperature with a thermal camera to ensure a safe operation. Cumulus-oocyte complexes were denuded by conventional MP with 40 IU/mL of hyaluronidase serving as control or by AHTs with a reduced amount of hyaluronidase (15 IU/mL). Piezo-ICSI was performed on both experimental and control groups. A triplicate of denudation and insemination experiments was performed. All embryos were monitored in a time-lapse incubator. Embryo developmental rates were compared by the chi-square test. Embryo morphokinetic timing as a continuous variable was compared by 1-way analysis of variance. Embryo transfers were performed on some blastocysts.

The procedural time for each denudation method was measured and compared. Fertilization, embryo development and morphokinetics, pregnancy, and live birth rate were compared between the control and experimental cohorts.

Facile noncontact denudation was achieved without any damage to oocyte. Acoustic induced fluidic shear was the main contributor to COC denudation. The average denudation time per oocyte decreased by 46% (15 seconds per oocyte for control vs. 8 seconds per oocyte for AHT) while using a lower concentration of hyaluronidase. Piezo-ICSI on oocytes processed by MP and AHTs resulted in comparable rates of survival (86.1% vs. 85.3%), fertilization (96.7% vs. 94.1%), and blastocyst (88.0% vs. 81.3%). Embryo morphokinetics for both experimental and control cohorts were comparable, showing no impact of sound waves on the embryo development. Eventual delivery rates were also comparable between the MP and AHT cohorts (51.3% vs. 55.4%).

Acoustohydrodynamic tweezers are used for contactless removal of the cumulus cells from the COCs before ICSI in an expedited, safe, and reliable manner. Embryo development outcomes confirm their safety and validate their potential for a comprehensive ICSI-on-chip device.

Intracytoplasmic sperm injection (ICSI), a type of assisted reproductive technology (ART), is offered as a treatment option for male factor infertility. Over the years, the indications for ICSI have been expanded, despite uncertainty about its benefits and harms compared to the conventional method of achieving fertilisation. Artificial oocyte activation (AOA), which can be performed by chemical, electrical or mechanical intervention, has been employed during ART ICSI treatment where there has been a history of low fertilization rate or total fertilization failure, and it has been reported to improve reproductive outcomes. It is important to evaluate the clinical effectiveness and safety of AOA in women undergoing ART ICSI treatment.

To evaluate the benefits and harms of artificial oocyte activation in women affected by infertility undergoing intracytoplasmic sperm injection treatment.

We searched the following electronic databases: the Cochrane Gynaecology and Fertility Group Specialised Register, CENTRAL, MEDLINE, Embase, ClinicalTrials.gov and WHO international Clinical Trials Registry Platform (8 August 2024). We also searched reference lists of relevant articles and contacted experts in the field.

Randomized controlled trials comparing artificial oocyte activation (AOA) (chemical, electrical or mechanical interventions) versus no intervention, placebo or another method of AOA in women undergoing ART.

We used methodological procedures as per Cochrane recommendations. We assessed the risk of bias in the included studies using ROB 2. The primary outcomes were live birth and miscarriage rates. We analyzed data using the risk ratio (RR) and a fixed-effect model. We assessed the certainty of the evidence by using GRADE criteria. We restricted the primary analyses to studies at low risk of bias.

We included a total of 20 studies, four of which were participant-based randomized trials with 743 participants. The remaining 16 were sibling-oocyte-model randomized studies. We based the main clinical findings of the current review on the participant-based RCTs, and we restricted our primary analysis to studies with a low risk of bias. Based on the one trial with 343 participants that we included in our primary analysis, the evidence is very uncertain about the effect of AOA on the live birth rate when compared to conventional ICSI without AOA in women undergoing ART ICSI (RR 1.97, 95% CI 1.29 to 3.01; one trial; 343 participants). For a typical clinic with a live birth rate of 18% following ART, the addition of AOA may result in live birth rates between 24% and 55%, but this evidence is very uncertain. The evidence is very uncertain about the effect of AOA on the miscarriage rate compared to conventional ICSI without AOA in women undergoing ART ICSI (RR 0.99, 95% CI 0.48 to 2.04; one trial; 343 participants). If the miscarriage rate was 9% following ART, addition of oocyte activation may result in miscarriage rates between 4% and 18%, but this evidence is very uncertain. The evidence is very uncertain about the effect of AOA on the clinical pregnancy rate compared to conventional ICSI without AOA in women undergoing ART ICSI (RR 1.67, 95% CI 1.20 to 2.32; one trial; 343 participants). The evidence is very uncertain about the effect of AOA on the multiple pregnancy rate per participant compared to conventional ICSI without AOA in women undergoing ART ICSI (RR 1.91, 95% CI 0.48 to 7.67; one trial; 343 participants). The evidence is very uncertain about the effect of AOA on the total fertilization failure rate compared to conventional ICSI without AOA in women undergoing ART ICSI (RR 0.05, 95% CI 0.01 to 0.40; one trial; 343 participants). When we stratified our analysis according to various infertility factors, we found low-certainty evidence that in couples undergoing ICSI treatment who have had a history of low or no fertilization, AOA may help improve the live birth rate while making little or no difference to the miscarriage rate. Further research is needed to confirm or refute this finding. None of the trials reported congenital anomalies (birth defects) as an outcome. Lack of short- or long-term safety data is an important limitation of the review and of the trials in this field. We did not find any trials that compared two different methods of oocyte activation.

We are uncertain about the effect of AOA on the live birth and miscarriage rates in women undergoing ART ICSI. In the subpopulation of those who have had a previous history of low or no fertilization, AOA may result in an increase in the live birth rate when compared to conventional ICSI without AOA, while making little or no difference to the miscarriage rate. There was considerable variation in the protocols used for chemical AOA, which affects the generalizability of the findings. Due to the very low to low certainty of evidence, the results should be interpreted with caution.

Classical preimplantation embryo culture is performed in static fluid environments. Whether a dynamic fluid environment, like the fallopian tube, is beneficial for embryo development remains to be determined across mammalian species. Objectives of these proof-of-concept studies were to determine if controllable dynamic microfluidic culture would enhance preimplantation murine, bovine, and human embryo development compared to static culture. This prospective randomized controlled trial tested static versus controlled dynamic culture of preimplantation mouse (n = 397), bovine (n = 242), and human (n = 512) zygotes to blastocyst stages with outcome measures of embryo cleavage, cellular fragmentation, apoptosis, and blastocyst conversion rates. Dynamic culture of mouse and bovine zygotes with microfluidics significantly improved embryo development. Mouse placental imprinted gene expression was significantly different between embryos derived in vivo, by static culture, and by dynamic culture. Using human sibling zygotes, this dynamic microfluidic culture system increased the number of blastomeres per cleavage-stage embryo, reduced cellular fragmentation or apoptosis, improved blastocyst conversion rates, and enhanced blastocyst developmental stages. In conclusion, species-specific longitudinal studies demonstrated that dynamic microfluidic culture significantly improved embryo development, independent of culture media composition, temperature, and gaseous environment. These cellular indicators represent improved embryo development that can translate into higher pregnancy rates in transgenics, domestic livestock and endangered species and treating human infertility.

Infertility is a highly prevalent disease affecting 1 in 6 couples worldwide. While there are numerous treatment options available to treat infertility, in vitro fertilization (IVF) is the most successful therapy available. IVF traditionally involves superovulation of the ovaries followed by ultrasound (US)-guided oocyte retrieval, fertilization of oocytes in vitro, and culture of resultant embryos. Embryo(s) are then either transferred into the uterus or cryopreserved. There have been tremendous advances in each step of this process over the past 40 years, however, much work still remains to be done. This review discusses the history and current techniques involved in oocyte fertilization and embryo culture. Highlighted areas include conventional insemination and intracytoplasmic sperm injection (ICSI), embryo culture, and laboratory assessment. Areas of past and present research including time-lapse imaging and in vitro maturation are also discussed.

This study compared the outcomes of conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in patients with polycystic ovarian syndrome (PCOS), tubal factor (TF) infertility, and unexplained infertility whose partners had normal semen parameters.

This retrospective study included 360 couples diagnosed with infertility involving PCOS (n=157), unexplained infertility (n=140), and TF infertility (n=63). Sibling oocytes were randomly assigned to undergo ICSI or conventional IVF insemination. The fertilization rate and embryo morphology were evaluated as outcomes.

Retrieved cumulus-oocyte complexes from patients with PCOS (2,974), unexplained infertility (1,843), and TF infertility (844) were split and inseminated by conventional IVF and ICSI respectively. In comparison to the ICSI method, the conventional IVF approach was linked to a significantly higher fertilization rate in groups with PCOS (68.81% vs. 77.49%), unexplained infertility (67.62% vs. 78.84%), and TF issues (69.23% vs. 78.63%) (p<0.05). The proportion of embryos with grade A produced by the conventional IVF method was significantly higher than that produced using the ICSI method in the PCOS and unexplained infertility groups (p<0.05). Additionally, the percentage of grade B embryos produced with the ICSI method was significantly higher than that produced with the conventional IVF method in PCOS patients (p=0.002).

Our results indicated that the conventional IVF method was associated with higher zygote production and a higher proportion of grade A embryos when all infertile groups were evaluated together. Thus, ICSI is not suggested for patients with these causes of infertility if their partner has normal semen parameters.

Which independent factors influence ICSI outcomes in patients with complete azoospermia factor c (AZFc) microdeletions?

In patients with complete AZFc microdeletions, the sperm source, male LH, the type of infertility in women, and maternal age are the independent factors associated with ICSI outcomes.

AZF microdeletions are the second most prevalent factor contributing to infertility in men, with AZFc microdeletions being the most frequently affected locus, accounting for 60-70% of all cases. The primary clinical phenotypes are oligoasthenozoospermia and azoospermia in patients with complete AZFc microdeletions. These patients can achieve paternity through ICSI using either testicular (T-S) or ejaculated (E-S) spermatozoa. With aging in men with AZFc microdeletions, oligoasthenozoospermia or severe oligozoospermia may gradually progress to azoospermia.

In this retrospective cohort study, the independent factors associated with the outcomes of 634 ICSI cycles in 634 couples with the transfer of 1005 embryos between February 2015 and December 2023 were evaluated. The analysis included 398 ICSI cycles in 398 couples using E-S and 236 ICSI cycles in 236 couples using T-S; all men had complete AZFc microdeletions.

The inclusion criteria were as follows: (i) men had complete AZFc microdeletions and (ii) the couple underwent ICSI treatment using T-S or E-S. The exclusion criteria were as follows: (i) cycles involving frozen-thawed oocytes; (ii) cycles in which all fresh embryos were frozen and not transferred; (iii) cycles lost to follow-up; and (iv) multiple ICSI cycles, apart from the first cycle for each couple. The primary outcome was the cumulative live birth rate per ICSI cycle, whereas the secondary outcomes were the clinical pregnancy rate per ICSI cycle, fertilization rate, and the no-embryo-suitable-for-transfer cycle rate (NESTR). Moreover, the maternal and neonatal outcomes were analyzed. Continuous variables showing non-normal distributions were expressed as median and interquartile range and were analyzed using the Kruskal-Wallis test. Categorical variables were expressed as percentages and were analyzed using the χ2 or Fisher's exact test. Linear and logistic regression models were constructed to assess the independent factors associated with ICSI outcomes.

The T-S group exhibited inferior ICSI outcomes than the E-S group, marked by significantly reduced rates of cumulative live birth, clinical pregnancy, fertilization, high-quality embryos, blastocyst formation, and implantation, with higher NESTRs. However, the miscarriage rate and neonatal outcomes did not significantly differ between the groups. Multivariate linear regression analysis demonstrated that reduced fertilization rates were significantly associated with T-S use (adjusted β, -0.281; 95% CI, -0.332 to -0.229). Multivariate logistic regression demonstrated that increased NESTRs were significantly associated with T-S use (adjusted odds ratio (OR), 4.204; 95% CI, 2.340-7.691), along with uterine anomaly in women (adjusted OR, 2.853; 95% CI, 1.053-7.718), infertility in women with multiple etiologies (adjusted OR, 11.118; 95% CI, 2.034-66.508), and advanced maternal age (adjusted OR, 1.138; 95% CI, 1.029-1.263). The use of T-S (adjusted OR, 0.318; 95% CI, 0.188-0.528), uterine anomaly in women (adjusted OR, 0.263; 95% CI, 0.058-0.852), and increased maternal age (adjusted OR, 0.877; 95% CI, 0.801-0.958) were also associated with decreased clinical pregnancy rates per ICSI cycle. Likewise, lower cumulative live birth rates were associated with T-S use (adjusted OR, 0.273; 95% CI, 0.156-0.468), male LH levels (adjusted OR, 0.912; 95% CI, 0.837-0.990), uterine anomaly (adjusted OR, 0.101; 95% CI, 0.005-0.529), and increased maternal age (adjusted OR, 0.873; 95% CI, 0.795-0.958). No significant differences were observed in the maternal and neonatal outcomes between both groups.

The study was based on a single-center, retrospective cohort design. The molecular diagnosis of AZFc microdeletions was reliant on loci sY254 and sY255 according to the European Academy of Andrology and European Molecular Genetics Quality Network guidelines. While our findings were based on the clinical phenotypes and laboratory parameters, the abnormalities in the genetic profiles of spermatogenesis and early embryonic development in patients between the T-S and E-S groups have not yet been elucidated.

Our results offer important insights into the independent factors that influence ICSI outcomes in patients with complete AZFc microdeletions. ICSI using E-S is a more favorable therapeutic option for younger patients with AZFc microdeletions and with sperm present in their ejaculate. This study highlights a new direction to investigate the molecular and phenotypic differences between the T-S and E-S groups, which may contribute to the diagnosis and treatment of complete AZFc microdeletions.

This study was supported by Capital's Funds for Health Improvement and Research (2022-2-4094), Beijing Natural Science Foundation (7232203, 7242164), National Key Research and Development Program (2021YFC2700200, 2023YFC2705600), National Natural Science Foundation of China (82301889), Peking University Third Hospital Innovation Transformation Fund (BYSYZHKC2023103), Peking University Third Hospital Clinical Cohort Construction Project (BYSYDL2023016), and Young Elite Scientists Sponsorship Program by CAST (2023QNRC001). None of the authors have any competing interests to declare.

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Intracytoplasmic sperm injection (ICSI) is currently being abused as it is also frequently employed in the absence of a frank male cause of infertility. There is growing and robust evidence showing that the use of ICSI should be limited to couples with male infertility. For all other causes of infertility, the procedure does not increase the chance of live birth, may increase malformations in newborns and requires more resources than conventional IVF. In addition, the use of ICSI may impact the sex ratio at birth, favouring female infants. Even if this effect may not be targeted as a gender biased procedure, it should be seen as a wake-up and warning call. It reminds us that ICSI is a 'non-natural' invasive technique that circumvents natural sperm selection mechanisms. Overall, a policy of indiscriminate use of ICSI is not justified and a plea is made for a return to a higher use of conventional IVF. Fear about failed fertilization or the use of preimplantation genetic testing for aneuploidies may represent barriers to the need to lower ICSI use and should be overcome.

Currently, assisted reproduction clinics employ various sperm selection techniques to identify the best sperm for fertilization. However, these techniques may not assess crucial sperm traits that can substantially impact embryonic quality. To address this, we propose analyzing diverse histone modifications as potential markers of sperm functionality and success in assisted reproduction techniques.

Cross-sectional pilot study including infertile male patients attending an infertility clinic in CABA, Argentina between April and August 2019 was performed. We used immunofluorescence techniques to evaluate post-translational modifications of histones in sperm and established correlations with in vitro fertilization outcome and embryo quality.

Our findings indicate a negative correlation between H3K4me3 and H3K4me2 marks and fertilization rate and showed a positive correlation of this parameter with H3K9me mark. In addition, there was a positive correlation between H3K27me3 and good embryo quality.

This pilot study proposes a non-invasive strategy to predict embryo quality by analyzing spermatozoa prior to fertilization. The assessment of histone post-translational modifications in sperm samples could provide useful information for the recognition of epigenetic marks that could predict the health of the embryo of an assisted fertilization treatment.

Nowadays, the most adopted technique to address infertility problems is in vitro fertilisation (IVF). However, its success rate is limited, and the associated procedures, known as assisted reproduction technology (ART), suffer from a lack of objectivity at the laboratory level and in clinical practice. This paper deals with applications of Artificial Intelligence (AI) techniques to IVF procedures. Artificial intelligence is considered a promising tool for ascertaining the quality of embryos, a critical step in IVF. Since the oocyte quality influences the final embryo quality, we present a systematic review of the literature on AI-based techniques used to assess oocyte quality; we analyse its results and discuss several promising research directions. In particular, we highlight how AI-based techniques can support the IVF process and examine their current applications as presented in the literature. Then, we discuss the challenges research must face in fully deploying AI-based solutions in current medical practice. Among them, the availability of high-quality data sets as well as standardised imaging protocols and data formats, the use of physics-informed simulation and machine learning techniques, the study of informative, descriptive yet observable features, and, above all, studies of the quality of oocytes and embryos, specifically about their live birth potential. An improved understanding of determinants for oocyte quality can improve success rates while reducing costs, risks for long-term embryo cultures, and bioethical concerns.

We aimed to compare the success rate of spermatozoa retrieval through microscopic testicular sperm extraction (mTESE) in non-obstructive azoospermic (NOA) men with a solitary testis with that of mTESE in NOA men with bilateral testes and the parameters affecting these rates.

A retrospective cross-sectional study of factors contributing to infertility in NOA patients with a solitary testis and men with bilateral testes was carried out. In this multicenter study, 74 patients with NOA with a solitary testis were matched with 74 patients with bilateral testes in terms of age, duration of infertility, and volume of the solitary testis from 2770 patients with NOA with bilateral testes. Hormonal parameters, presence of varicocele, history of varicocelectomy, history of undescended testis and karyotype analysis results were compared.

Spermatozoa were obtained from 40 (54.1%) patients with a solitary testis and 42 (56.76%) patients with bilateral testes. No differences were found regarding age, duration of infertility, or mean testicular volume between patients with a solitary testis and patients with bilateral testes. When serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were compared regardless of sperm retrieval status, it was observed that both levels were greater in the group of patients with a solitary testis (p < 0.01). Patients with solitary and bilateral testes from whom spermatozoa were obtained had larger testes than those from whom spermatozoa could not be obtained (p < 0.05). Similarly, the serum levels of FSH and LH were significantly greater in patients with a solitary testis than in those with bilateral testes (p < 0.05).

To the best of our knowledge, this is the first study in the literature to evaluate the parameters that influence mTESE outcome in NOA patients with a solitary testis and NOA patients with bilateral testes. Greater testicular volume was found to positively affect spermatozoa retrieval for patients with a solitary testis. The higher levels of FSH and LH in patients with a solitary testis than in patients with bilateral testes of similar testicular volume may be due to a compensatory mechanism developed by the hypothalamic-pituitary-gonadal axis. The fact that these hormones are higher in patients with a solitary testis does not mean that the number of spermatozoa obtained through mTESE will be decreased.

To explore the application possibility of macrocephalic sperm from a patient with 100% macrocephalic sperm and AURKC gene variations.

We diagnosed a case of macrozoospermia with 100% macrocephalic sperm and 39.5% multi-tailed spermatozoa by morphological analysis. Whole-exome sequencing (WES) was used for the patient and his wife. Sanger sequencing technique was used to verify the AURKC mutations in the patient's parents and his offspring. Sperm's ploidy was tested by flow cytometry. The couple asked for intra-couple ART therapy.

The patient presented novel compound heterozygous AURKC mutations (c.434C > T, c.497A > T) by WES. Sanger sequencing validation showed that variant of c.434C > T was observed in his father and c.497A > T was observed in his mother. Flow cytometry revealed that there existed a certain proportion of haploid sperm. Macrocephalic spermatozoa whose heads were smaller than the diameter of injection needle were selected for microinjection. A singleton pregnancy was achieved after embryo transfer. Prenatal diagnosis revealed that the fetus had normal chromosomal karyotype. Sanger sequencing technique showed that the fetus carried a c.434C > T mutation in one AURKC allele. A 3730 g healthy male fetus was delivered at term.

Our study reported a successful live birth from a patient with definite AURKC gene variants and may provide insights for such patients to choose donor sperm or their own sperm.

Embryo development is stimulated by calcium (Ca2+) signals that are generated in the egg cytoplasm by the fertilizing sperm. Eggs are formed via oogenesis. They go through a cell division known as meiosis, during which their diploid chromosome number is halved and new genetic combinations are created by crossing over. During formation the eggs also acquire cellular components that are necessary to produce the Ca2+ signal and also, to support development of the newly formed embryo. Ionized calcium is a universal second messenger used by cells in a plethora of biological processes and the eggs develop a "toolkit", a set of molecules needed for signaling. Meiosis stops twice and these arrests are controlled by a complex interaction of regulatory proteins. The first meiotic arrest lasts until after puberty, when a luteinizing hormone surge stimulates meiotic resumption. The cell cycle proceeds to stop again in the middle of the second meiotic division, right before ovulation. The union of the female and male gametes takes place in the oviduct. Following gamete fusion, the sperm triggers the release of Ca2+ from the egg's intracellular stores which in mammals is followed by repetitive Ca2+ spikes known as Ca2+ oscillations in the cytosol that last for several hours. Downstream sensor proteins help decoding the signal and stimulate other molecules whose actions are required for proper development including those that help to prevent the fusion of additional sperm cells to the egg and those that assist in the release from the second meiotic arrest, completion of meiosis and entering the first mitotic cell division. Here I review the major steps of egg formation, discuss the signaling toolkit that is essential to generate the Ca2+ signal and describe the steps of the signal transduction mechanism that activates the egg's developmental program and turns it into an embryo.

Assisted reproductive technologies are an emerging field in equine reproduction, with species-dependent peculiarities, such as the low success rate of conventional IVF. Here, the 'cumulome' was related to the developmental capacity of its corresponding oocyte. Cumulus-oocyte complexes collected from slaughterhouse ovaries were individually matured, fertilized by ICSI, and cultured. After maturation, the cumulus was collected for proteomics analysis using label-free mass spectrometry (MS)-based protein profiling by nano-HPLC MS/MS and metabolomics analysis by UPLC-nanoESI MS. Overall, a total of 1671 proteins and 612 metabolites were included in the quantifiable 'cumulome'. According to the development of the corresponding oocytes, three groups were compared with each other: not matured (NM; n = 18), cleaved (CV; n = 15), and blastocyst (BL; n = 19). CV and BL were also analyzed together as the matured group (M; n = 34). The dataset revealed a closer connection within the two M groups and a more distinct separation from the NM group. Overrepresentation analysis detected enrichments related to energy metabolism as well as vesicular transport in the M group. Functional enrichment analysis found only the KEGG pathway 'oxidative phosphorylation' as significantly enriched in the NM group. A compound attributed to ATP was observed with significantly higher concentrations in the BL group compared with the NM group. Finally, in the NM group, proteins related to degradation of glycosaminoglycans were lower and components of cumulus extracellular matrix were higher compared to the other groups. In summary, the study revealed novel pathways associated with the maturational and developmental competence of oocytes.

Intracytoplasmic sperm injection (ICSI) remains inefficient in cattle. One reason could lie in the injection of oocytes with sperm that have not undergone molecular changes associated with in vivo capacitation and fertilizing ability. This study aimed to enhance the efficiency of bovine intracytoplasmic sperm injection (piezo-ICSI) by employing fluorescent-activated cell sorting (FACS) to select the sperm population before injection based on capacitation markers. First, we evaluated the effects of incubating thawed sperm for 2 hours with different capacitating inductors: heparin, methyl-beta-cyclodextrin (MβCD), and dibutyryl cyclic AMP (dbcAMP), alone or in combinations in a basal capacitating (C) medium (Sp-TALP). Sperm capacitation and quality markers were evaluated by flow cytometry, revealing heparin as the most effective inducer of sperm capacitation changes. It, therefore, this treatment was chosen as the sperm pretreatment for FACS-piezo-ICSI. Two cell populations showing high capacitating levels (Heparin-HCL) and low capacitating levels (Heparin-LCL) of the markers associated with sperm capacitation i(Ca2+) levels and acrosome integrity were selected by FACS and used for sperm injection. Pronuclear formation was significantly higher when ICSI was performed with Heparin-HCL sperm than with Heparin-LCL and the control group (Heparin unsorted) groups (50 %, 10 %, and 20 %, respectively). Furthermore, injecting Heparin-HCL sperm resulted in a higher blastocyst rate (22.5 %) than Heparin-LCL (10 %) and the control group (15.2 %). In conclusion, heparin treatment effectively induced changes associated with sperm capacitation. The combination of Heparin-HCL treatment and FACS enabled precise selection of capacitated sperm before ICSI, enhancing the efficiency of this technology in the bovine species.

Current infertility treatment strategies focus on mature gametes, leaving a significant proportion of cases with gamete progenitors that stopped complete differentiation. On the other hand, recent advancements in next-generation sequencing have identified many candidate genes that may promote maturation of germ cells. Although gene therapy has shown success in mice, concerns about the integration of DNA vectors into oocytes hinder clinical applications. Here, we present the restoration of fertility in female mice through Sendai virus (SeV)-mediated RNA delivery. Ovaries lacking Kitl expression exhibit only primordial follicles due to impaired signaling to oocytes expressing the KIT tyrosine kinase. Despite SeVs being immunogenic and larger than the blood-follicle barrier, the administration of Kitl-expressing SeVs reinitiated oogenesis in genetically infertile mice that have only primordial follicles, resulting in the birth of normal offspring through natural mating. This virus also effectively addressed iatrogenic infertility induced by busulfan, a widely used cancer chemotherapy agent. Offspring born through SeV administration and natural mating displayed normal genomic imprinting patterns and fertility. Since SeVs pose no genotoxicity risk, the successful restoration of fertility by SeVs represents a promising approach for treating congenital infertility with somatic cell defects and protecting fertility of cancer patients who may become infertile due to loss of oocytes during cancer therapy.

To compare cumulative live birth rate (CLBR) and cost-effectiveness of intracytoplasmic sperm injection (ICSI) vs. conventional in vitro fertilization (cIVF).

Retrospective cohort study of cycles reported to the Society for Assisted Reproductive Technology Clinic Outcomes Reporting System.

Society for Assisted Reproductive Technology (SART) member IVF clinics in the United States.

Patients with unexplained infertility who underwent first autologous retrieval cycles between January 2017 and December 2019 with linked fresh and frozen embryo transfers through December 2021.

ICSI vs. cIVF.

The primary outcome was CLBR, defined as ≤1 live birth from a retrieval cycle and all linked embryo transfers. Secondary outcomes included two pronuclear (2PN) per oocyte retrieved, miscarriage rate, and total number of transferred or frozen embryos per 2PN. Subsamples with and without preimplantation genetic testing for aneuploidy (PGT-A) were analyzed. Outcomes were adjusted for age, body mass index, number of oocytes retrieved, length of follow-up, and clinic ICSI use rate.

A total of 18,805 patients with unexplained infertility were included. No difference in CLBR was found among cycles without genetic testing (54.4% ICSI vs. 57.5% cIVF) and with PGT-A (47.6% ICSI vs. 51.8% cIVF). Intracytoplasmic sperm injection cycles without genetic testing had a higher miscarriage rate (16.4% vs. 14.4%) but no difference was seen in cycles with PGT-A (13.9% ICSI vs. 13.2% cIVF). Intracytoplasmic sperm injection cycles had a significantly lower ratio of 2PN per oocyte retrieved without genetic testing (59.7% vs. 60.9%) and with PGT-A (63.3% vs. 65.8%). The ratio of embryos transferred or frozen per 2PN was not significantly different in cycles without genetic testing (49.4% vs. 49.6%) or with PGT-A (54.2% vs. 55.2%). Total fertilization failure occurred in 216 patients (4%) who underwent cIVF and in 153 patients (1.1%) who used ICSI.Compared with cIVF alone, an estimated additional $11,011,500 was charged to patients for ICSI without genetic testing and $9,010,500 was charged to patients for ICSI with PGT-A over 2 years by Society for Assisted Reproductive Technology clinics. On the basis of total fertilization failure rates, 35 patients would require treatment with routine ICSI to avoid a single cycle of total fertilization failure with cIVF.

Routine use of ICSI in unexplained infertility is not warranted due to the additional cost and lack of CLBR benefit.

To assess the impact of maternal age on the association between maternal basal FSH and aneuploidy.

A retrospective study including data from 1749 blastocysts diagnosed as euploid or aneuploid by PGT-A (preimplantation genetic testing for aneuploidy). Aneuploidy incidence was compared between embryos from mothers with high vs. low basal FSH levels (above and below the group median, respectively) in total, pre-AMA (advanced maternal age; < 35 years, 198 embryos) and AMA (≥ 35 years, 1551 embryos) patient groups, separately. To control for the interference of potentially confounding variables, the association between aneuploidy and high basal FSH levels was assessed by multivariate logistic analysis in overall, pre-AMA and AMA patient groups.

Overall, aneuploidy rate was 9% higher (p = 0.02) in embryos from patients with high basal FSH (63.7%) compared to those with low basal FSH (58.4%). In the pre-AMA subgroup, aneuploidy incidence was 35% higher (p = 0.04) in embryos from patients with high basal FSH (53.5%) compared to those with low basal FSH (39.4%). Differently, aneuploidy occurrence did not vary between embryos from AMA patients with low (61.0%) and high (64.8%) basal FSH (p = 0.12). The multivariate analysis revealed that, in pre-AMA embryos, the association between aneuploidy occurrence and high basal FSH is independent of potential confounding variables (p = 0.04).

Maternal basal FSH values are associated with embryo aneuploidy in pre-AMA but not in AMA patients. The present findings suggest that basal FSH is a useful parameter to assess aneuploidy risk in pre-AMA patients and reinforce the hypothesis that excessive FSH signalling can predispose to oocyte meiotic errors.

This conference celebrates the 40th anniversary of AETE. Over the past 40 years, AETE has served as a forum for scientists, practitioners, and students working in assisted animal reproduction in livestock species. AETE conferences have reflected developments in the field, from basic to applied science, as well as regulatory changes in assisted animal reproduction practices. Europe has led the way in these developments for many years, progressing from artificial insemination, embryo transfer, and cryopreservation to semen sexing, in vitro production of embryos, cloning by nuclear transfer, genomic selection, and the rescue of highly endangered species. These significant contributions were made possible by the support of funding agencies, both at the national and European levels, promoting cooperation between scientists and practitioners. Assisted reproduction, and animal breeding more generally, face opposition from various groups, including animal rights activists, vegetarians, proponents of organic farming, environmentalists, certain political parties, and increasing regulatory burdens. These challenges seriously affect funding for scientific research, the work of practitioners, and the breeding industry as a whole. It is crucial to invest time and resources in communication to remind the public, politicians, and regulators of the achievements in this field and the contributions made to the food supply chain and the care of the rural and natural environment.

Non-obstructive azoospermia (NOA) is found in up to 15% of infertile men. While several causes for NOA have been identified, the exact etiology remains unknown in many patients. Advances in assisted reproductive technology, including intracytoplasmic sperm injection (ICSI) and testicular sperm retrieval, have provided hope for these patients. This review summarizes the chances of success with ICSI for NOA patients and examines preoperative factors and laboratory techniques associated with positive outcomes. Furthermore, we reviewed possible consequences for offspring by the use of ICSI with testicular sperm retrieved from NOA patients and the interventions that could potentially mitigate risks. Testicular sperm retrieved from NOA patients may exhibit increased chromosomal abnormalities, and although lower fertilization and pregnancy rates are reported in NOA patients compared to other forms of infertility, the available evidence does not suggest a significant increase in miscarriage rate, congenital malformation, or developmental delay in their offspring compared to the offspring of patients with less severe forms of infertility or the offspring of fertile men. However, due to limited data, NOA patients should receive specialized reproductive care and personalized management. Counseling of NOA patients is essential before initiating any fertility enhancement treatment not only to mitigate health risks associated with NOA but also to enhance the chances of successful outcomes and minimize possible risks to the offspring.

Male infertility is a pervasive global reproductive challenge, primarily attributed to a decline in semen quality. Addressing this concern, there has been a growing focus on spermatozoa sorting in assisted reproductive technology. This study introduces a groundbreaking development in the form of a thermotaxis and rheotaxis microfluidic (TRMC) device designed for efficient motile spermatozoa sorting within a short 15-min timeframe. The TRMC device mimics the natural sperm sorting mechanism of the oviduct, selecting spermatozoa with superior motility and DNA integrity. The experimental outcomes demonstrate a remarkable enhancement in the percentage of progressive spermatozoa following sorting, soaring from 3.90% to an impressive 96.11% when subjected to a temperature decrease from 38 °C to 35 °C. Notably, sperm motility exhibited a substantial 69% improvement. The TRMC device exhibited a commendable recovery rate of 60.93%, surpassing current clinical requirements. Furthermore, the sorted spermatozoa displayed a notable reduction in the DNA fragmentation index to 6.94%, signifying a substantial 90% enhancement in DNA integrity. This remarkable advancement positions the TRMC device as highly suitable for applications in in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), offering a promising solution to male infertility challenges.

To study whether severe male factor infertility (SMF), reflected by oligozoospermia, impacts embryo morphokinetic behavior in low-prognosis women as stratified by the Patient-Oriented Strategies Encompassing IndividualizeD Oocyte Number (POSEIDON) criteria.

Cohort study.

Private university-affiliated in vitro fertilization center.

A total of 10,366 injected oocytes from 2,272 women who underwent intracytoplasmic sperm injection cycles between March 2019 and April 2022.

Patients were divided into 8 groups according to the POSEIDON criteria (1-4) and the presence or absence of SMF. A control group of normoresponder patients was included. Kinetic markers from the point of insemination were recorded in the EmbryoScope incubator.

Morphokinetic milestones and intracytoplasmic sperm injection clinical outcomes.

Embryos from patients in the POSEIDON 1 group showed significantly slower timing to pronuclear appearance, timing to pronuclear fading (tPNf), timing to 2 (t2), 3 (t3), 4 (t4), 6 (t6), and 7 (t7) cells than those from the control group. Known Implantation Diagnosis Score ranking was significantly different between the SMF and non-SMF (nSMF) subgroups in both POSEIDON 1 as well as control groups. Embryos from patients in the POSEIDON 2 group showed significantly slower timing to pronuclear appearance, t4, t6, t7, timing to 8 cells (t8), and timing to morulae than those from the control group. Embryos in the POSEIDON 2 SMF subgroup took longer than those in the POSEIDON 2 nSMF subgroup and those in both control subgroups to achieve tPNf, t2, t3, timing to 5 cells (t5), timing to start blastulation, and timing to blastulation. Known Implantation Diagnosis Score ranking was significantly different between the SMF and nSMF subgroups in both POSEIDON 2 as well as control groups. Embryos from patients in the POSEIDON 3 group showed significantly slower t8 and duration of the second cell cycle (t3-t2) than those from the control group. Known Implantation Diagnosis Score ranking was significantly different across the subgroups. Embryos derived from patients in the POSEIDON 4 group showed significantly slower tPNf, t2, t3, t4, t5, t6, t7, t8, timing to complete t4-t3 synchronous divisions, and timing to complete t8-t5 synchronous divisions than those from the control group. Known Implantation Diagnosis Score ranking was significantly different between the SMF and nSMF subgroups in both POSEIDON 4 as well as control groups. Irrespective of sperm quality, clinical outcomes significantly improved in the control subgroups compared with those in the POSEIDON 2 and 4 subgroups.

Embryos in the SMF groups presented lower Known Implantation Diagnosis Score ranking than those in the nSMF groups in both POSEIDON 1-4 and control groups, suggesting that cumulative differences result in worse morphokinetic development when the algorithm is used.

In assisted reproductive technology (ART), the choice between intracytoplasmic sperm injection (ICSI) and conventional in vitro insemination (IVF) remains a pivotal decision for couples with female or unexplained infertility. The hypothesis that ICSI may not confer significant improvements in live birth rates in the absence of a male infertility factor was explored in this study.

This was a retrospective collection of data recorded by the Human Fertilisation and Embryology Authority (HFEA) in the UK from 2005 to 2018 and analysed through regression analysis models on both the entire dataset and a matched-pair subset. First fresh ART cycles were analysed according to the insemination technique in order to compare live birth as the main outcome. Cycles were included if complete information regarding infertility cause, female age, number of oocytes retrieved, allocation to ICSI or IVF, and treatment outcome in terms of live birth was available. Matching was performed at a 1:1 ratio between IVF and ICSI cycles according to the cause of infertility, female age, number of oocytes, and year of treatment.

This study, based on 275,825 first cycles, revealed that, compared with IVF, ICSI was associated with higher fertilization rates and lower cycle cancellations rates. However, ICSI was associated with a lower chance of implantation and live birth than IVF in cycles with female-only infertility: in the entire dataset, the adjusted odds of having a live birth decreased by a factor of 0.95 (95% CI 0.91-0.99, p = 0.011), while in the matched-pair analyses it decreased by a factor of 0.91 (95% CI 0.86-0.96, p = 0.003) using ICSI compared to IVF. For unexplained infertility cycles, the adjusted odds ratios for live birth in ICSI compared to IVF cycles were 0.98 (95% CI 0.95-1.01) in the entire dataset and 0.97 (95% CI 0.93-1.01) in the matched-pair analysis.

Compared with IVF, ICSI was associated with a reduction in live births when ART was indicated due to female-only factors. Additionally, no significant improvements were associated with the use of ICSI in cycles with unexplained infertility. Our findings impose a critical reevaluation regarding the use of ICSI over IVF for cases with female-only factors and unexplained infertility.

To examine the possible synergic effect of spindle view-assisted intracytoplasmic sperm injection (SV-ICSI) with assisted oocyte activation (AOA) for low fertilization rate.

A single-center retrospective study from 2019/09-2023/06, a total of 47 patients, autologous IVF cycle, and low fertilization rate history, including control group (SV-ICSI, 33 patients) and intervention group (AOA-SV-ICSI, 14 patients), comparing fertilization rate, blastocyst formation rate, and clinical pregnancy rate.

The blastocyst formation rate was significantly higher (p = 0.020) in the AOA-SV-ICSI group than in the SV-ICSI group. The fertilization rate (P = 0.468) and clinical pregnancy rate (p = 0.057) were non-significant between groups.

The AOA-SV-ICSI group's blastocyst formation rate significantly improved in patients with previous low fertilization rates, which might help them obtain more useable embryos for further embryo implantation.

Fertilization failure often occurs in conventional IVF cycles, and day 1 rescue ICSI is frequently recommended. In this study, the effect of rescue ICSI on obstetrical and neonatal outcomes after a single blastocyst transfer in vitrified-warmed cycles is evaluated.

This cohort study was a retrospective analysis of 703 vitrified-warmed single blastocyst transfers and 219 singletons in the r-ICSI group compared with 11,611 vitrified-warmed single blastocyst transfers in the IVF/ICSI and 4472 singletons in the IVF/ICSI group, respectively, and patients just undergoing their first IVF treatments were included in this study. Pregnancy rate (PR), live birth rate (LBR), and singleton birthweight were the primary outcome measures. Multiple linear regression analysis and logistic regression analysis were performed to evaluate the possible relationship between obstetrical and neonatal outcomes and fertilization method (including IVF, ICSI, and r-ICSI) after adjusting for other potential confounding factors.