Male Factor Infertility Patients Research Articles

P-056 NOA-SERA: combined autologous progenitor cells and platelet-derived growth factors for male factor infertility due to non-obstructive azoospermia and severe oligoasthenoteratozoospermia (OATS) – a case series

Abstract Study question Can NOA-SERA be utilizing autologous progenitor cells and platelet-derived growth factors,effectively address male factor infertility in non-obstructive azoospermia (NOA) and severeoligoasthenoteratozoospermia (OATS) Summary answer Intra-testicular NOA-SERA transplantation of autologous progenitor cells andgrowth factors shows promise in improving spermatogenesis and fertilization potential,presenting potential breakthrough for male factor infertility. What is known already Male infertility affects 10–15% of couples, with non-obstructive azoospermia (NOA)diagnosed in 60% of azoospermic men. Current interventions, including platelet-rich plasma and stem cells, show variable success. This study introduces a novel protocol, NOA-SERA, combining autologous mobilized progenitor cells and platelet-derived growth factors to restore spermatogenesis. Despite advancements in assisted reproduction, male factor infertility due to primary NOA remains challenging. This case series explores the safety and efficacy of NOA-SERA, avoiding in vitro breeding risks and emphasizing intratesticular transplantation’s safety and efficacy Study design, size, duration his case series (March 2019–November 2023) includes 7 male factor infertility patients,consisting of 4 non-obstructive azoospermic NOA, 1 cryptozoospermia and 2 severe oligoaesthenoteratospermic (OATS, strongly desirous of pregnancy with self-gamete. All patients received NOA-SERA following written consent. One-year follow-up included hormonal profiling, semen analysis, and testicular biopsy assessment Participants/materials, setting, methods Seven men, with normal karyotype, underwent NOA-SERA due to male factor infertility. This intervention involved intra-testicular transplantation of autologous CD34+/CD133+ enriched peripheral blood mononuclear stem cells (MPB-MNCs) combined with platelet derived growth factors. The procedure, avoiding in vitro breeding and manipulation, prioritized safety and efficacy. Follow-up for one year included hormonal profile, semen analysis, testicular biopsy assessments, clinical pregnancy (b-hCG), sustained implantation (>8 weeks), and live birth rates were followed. Main results and the role of chance Analysis of NOA-SERA treatment in this case series revealed positive outcomes in 70% of patients, showing increased testicular size, elevated testosterone, and reduced FSH levels. In our study, out of 4 NOA, 2 NOA remained NOA, 1 progressed to OATS, 1 progressed to normal semen analysis after 3 months. Couple conceived with intrauterine insemination (IUI) first pregnancy and they also had a subsequent natural conception. 1 Crypto progressed to OATS and 1 severe OATS remained the same and one progressed to OATS. Although no appearance of sperm in the ejaculate was observed, 2 patients displayed sperms in Testicular Sperm Aspiration (TESA) sufficient for ICSI. Couples undergoing IVF therapy post-biopsy achieved successful fertilization in two cases, leading to positive pregnancies and sustained implantation and livebirth of healthy full-term babies. Post-procedure FSH was slightly lower than pre-procedural levels, indicating hormonal improvement without observed therapy-related deterioration. The novelty lies in the intratesticular transplantation of mobilized purified autologous CD34+/CD133+ enriched MPBMNCs with platelet-derived growth factors, avoiding in vitro risks.” To summarise, 3 out of 7 achieved successful pregnancy, 2 NOA with ICSI, 1 NOA progressed to normal semen analysis and pregnancy with IUI Limitations, reasons for caution Despite interesting results, limitations include the self-controlled case series design and asmall patient cohort, lacking a comparison to a control group. Future studies should include more patients, stratified for age, aetiology, and other risk factors, to further elucidate this intervention’s mechanism of action Wider implications of the findings NOA-SERA demonstrates promise in enhancing spermatogenesis and fertilization potential, emphasizing necessity for larger randomized trials to ascertain clinical efficacy, particularly in subgroups of OATS&NOA patients. NOA-SERA emerges as safe, cost-effective adjuvant for optimizing spermatogenesis and improving IVF success and pregnancy outcomes in male factor infertility. Trial registration number Not applicable

#244 : Reproductive Outcomes of Conventional In Vitro Fertilization and Intracytoplasmic Sperm Injection in Patients with Non-Severe Male Infertility Across Poor and Different Suboptimal Ovarian Response Categories

Background and Aims: For non-severe male factor infertility patients with poor and sub-optimal ovarian response, are there any differences in reproduction outcomes between IVF (in vitro fertilization) and ICSI (intracytoplasmic sperm injection) methods? Methods: Retrospective cohort study. Patients with non-severe male infertility and poor ovarian response who underwent in vitro fertilization or intracytoplasmic sperm injection at the Center for Reproductive Medicine, Peking University Third Hospital between 2009 and 2019 were included in this study (n = 30,352). Results: In all groups, intracytoplasmic sperm injection cycles involved older patients and a longer duration of infertility, while body mass index, AMH, and bFSH levels were similar. Independent of the number of oocytes retrieved, intracytoplasmic sperm injection significantly increased the fertilization rate, while in vitro fertilization cycles showed a higher implantation rate, clinical pregnancy rate, and live birth rate. No differences were observed in most obstetric outcomes, including abortion, early abortion, multiple fetus, cesarean delivery, gestational age, and congenital malformations rates. However, in vitro fertilization cycles showed higher preterm delivery rates and lower birth weights in groups with 4-6, and 7-9 oocytes retrieved. A higher female/male infant ratio was observed in intracytoplasmic sperm injection cycles when 4-6 oocytes were retrieved. Conclusions: In patients with non-severe male infertility and poor or suboptimal ovarian response, even if intracytoplasmic sperm injection increased the fertilization rate, in vitro fertilization exhibited significant advantages in implantation, clinical pregnancy, and live birth rates. Our study indicates that poor ovarian response is not an indication for intracytoplasmic sperm injection in couples with non-severe male infertility.

P-782 The influence of microfluidic preparation of spermatozoa for ICSI (intracytoplasmic sperm injection) in an unselected IVF population on fertilization rate, embryo-quality and pregnancy rate

Abstract Study question Does the microfluidic technique as a new preparation option for sperm in ICSI during IVF have a better outcome in fertilization, blastocysts and pregnancy rate? Summary answer The patients that had the sperm preparation for ICSI with the microfluidic sperm selection had similar results as with the gradient centrifugation selection. What is known already Semen analysis is a poor predictor of the fertilization potential of spermatozoa and a male factor may contribute to poor outcomes of IVF procedure despite a normal semen analysis. Traditionally, sperm processing is accomplished by density gradient centrifugation or swim-up methods but this have been shown in some studies to increase reactive oxygen species and induce sperm DNA fragmentation. The microfluidic sperm selection (ZyMot- ICSI) is based on the selection of the spermatozoa with the lowest DNA fragmentation rate but studies do not prove better clinical outcomes after this method. Study design, size, duration This is a prospective randomized study carried out in a university level fertility clinic in Oradea, Romania from January 2022 to December 2022, including 239 patients in total. The patients that underwent IVF-ICSI procedures for various indications were randomized in two groups: the control group swim-up technique was used for sperm selection for ICSI during IVF and the study group with the ZyMot-ICSI (microfluidic sperm selection technique) was used. Participants/materials, setting, methods We conducted a prospective trial comparing 119 couples that were allocated to the classic swim-up technique (control group) and 120 couples that were allocated with the microfluidic technique was used (study group) in our university level clinic to go through IVF. Inclusion criteria were couples with known infertility requiring IVF and willing to participate in the study. Exclusion criteria were: severe oligoasthenospermia (concentration < 5x10^6 spermatozoa/ml or motility <10%), oocyte or sperm donation. Main results and the role of chance The statistical analysis showed that there is no significant difference between the fertilization rate (study vs control p = 0.37 with absolute difference -1.182, 95% CI (-7.2, 5.4)) or blastocyst rate (study vs control p = 0.88 with absolute difference -0.5 95% (-6.5, 6)) nor is there a difference in clinical pregnancy. Even if microfluidic preparation of spermatozoa did not seem to improve the results it may be utilized more broadly for standard IVF, intrauterine insemination and could also improve workflow, decrease intervention by laboratory personal and provide more consistent incubation conditions. Limitations, reasons for caution The population studied was inclusive with unselected IVF indications and did not attempt to isolate male factor infertility cases or patients with a history of elevated sperm DNA fragmentation. Wider implications of the findings Microfluidic sperm preparation performs similarly to density gradient centrifugation in sperm preparation for IVF in an unselected population. Trial registration number Not applicable

P-366 Unraveling the whole transcriptome profiles of receptive phase endometrium in infertile women

Abstract Study question Are there differential transcriptomic profiles of the receptive phase endometria in infertile women with different infertility diagnoses? Summary answer Endometrium-related pathologies such as endometriosis and RIF seemed to share similar endometrial molecular profiles in mid-secretory phase,while samples from unexplained infertility were similar to controls. What is known already Successful embryo implantation is orchestrated by the combination of a viable blastocyst, a receptive endometrium and a perfectly synchronized molecular dialogue between them. Endometrium-associated pathologies such as endometriosis and recurrent implantation failure (RIF) are suggested to hamper endometrial receptivity, but the molecular mechanisms affected in these and other infertility diagnoses such as unexplained infertility need further investigation. Previous transcriptome studies of the human endometrium have analysed different infertility diagnoses separately, but whether and to what extent receptive-phase endometrial transcriptome profiles vary between distinct infertility diagnoses is underexplored. Study design, size, duration A prospective, cross-sectional study was performed at the Reproductive Unit at the University Hospital between March 2019 and April 2021. Forty-five well-characterized infertile women (age 34.7±3.8 years, BMI 24.5±kg/m2) were recruited before starting any infertility treatment. Four groups according to different infertility diagnoses were established: endometriosis (n = 12), RIF (n = 14), unexplained infertility (n = 10), and male factor infertility as control group (n = 9). Participants/materials, setting, methods Endometrial biopsies were collected using Pipelle curette during mid-secretory phase (LH + 7) of the natural cycle. Total RNA was extracted and sequenced using Illumina NovaSeq 6000 technology. Raw sequencing data was quality assessed, pre-processed and quantified, and differential expression and enrichment analyses were performed using R software. Main results and the role of chance Differential expression analysis revealed 743, 587 and 140 differentially expressed genes (DEGs) in endometriosis, RIF and unexplained infertility, respectively, when compared to control women of the male factor infertility group. Among these, 31 genes exhibited consistent differential expression between all infertility groups. Regarding functional enrichment analyses, the detected DEGs were mainly related to immune response and inflammatory processes, supporting the involvement of these pathways in the impairment of endometrial receptivity. Interestingly, the comparison between different study groups showed similar transcriptome profiles in endometrium-associated pathologies such as endometriosis and RIF, while women with unexplained infertility had similar molecular profiles with the control group of male factor infertility. These study results indicate common molecular pathways in endometrial receptivity between infertile women with endometriosis and RIF, and between unexplained infertility and control male factor infertility patients. Limitations, reasons for caution Further research with bigger sample size is required to confirm these findings. Wider implications of the findings We identified transcriptome profiles associated with infertility diagnoses. Further, women with unexplained infertility tended to have similar molecular profiles with control women, meaning that other factors than endometrial receptivity issues could lead to their infertility. Our study helps to understand the molecular mechanisms underlying female infertility in different infertility diagnoses. Trial registration number not applicable

Integrative analysis of transcriptomic and metabolomic profiles reveals abnormal phosphatidylinositol metabolism in follicles from endometriosis-associated infertility patients.

Endometriosis is a common gynecological disorder that causes female infertility. Our recent research found that excessive oxidative stress in ovaries of endometriosis patients induced senescence of cumulus granulosa cells. Here, we analyzed the transcriptomic and metabolomics profiles of follicles in a mouse model of endometriosis and in patients with endometriosis and investigated the potential function of changed metabolites in granulosa cells. RNA-sequencing indicated that both endometriosis lesions and oxidative stress in mice induced abnormalities of reactive oxidative stress, steroid hormone biosynthesis, and lipid metabolism. The mouse model and women with endometriosis showed altered lipid metabolism. Nontargeted metabolite profiling of follicular fluid from endometriosis and male-factor infertility patients by liquid chromatography mass spectrometry identified 55 upregulated and 67 downregulated metabolites. These differential metabolites were mainly involved in steroid hormone biosynthesis and glycerophospholipid metabolism. Phosphatidylinositol (PI 16:0/18:2) was significantly elevated in follicular fluid from endometriosis patients compared with controls (p < 0.05), while lysophosphatidylinositol (LPI 18:2, 20:2, 18:1, 20:3 and 18:3) was reduced (p < 0.05). Upregulated PI and downregulated LPI correlated with oocyte retrieval number and mature oocyte number. LPI inhibited cellular reactive oxidative stress induced by hemin in granulosa cells. Cell proliferation inhibition, senescence, and apoptosis induced by hemin were partially reversed by LPI. Moreover, LPI administration rescued hemin blocking of cumulus-oocyte complex expansion and stimulated expression of ovulation-related genes. Transcriptomic Switching mechanism at 5' end of the RNA transcript sequencingand western blot revealed that LPI effects on granulosa cells were associated with its regulation of MAPK-ERK1/2 signaling, which was suppressed in the presence of hemin. In conclusion, our results revealed the dysregulation of lipid metabolism in endometriotic follicles. LPI may represent a novel agent for in vitro follicular culture that reverses the excessive oxidative stress from endometriotic lesions. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Microfluidic preparation of spermatozoa for ICSI produces similar embryo quality to density-gradient centrifugation: a pragmatic, randomized controlled trial.

Does processing of spermatozoa for IVF with ICSI by a microfluidic sperm separation device improve embryo quality compared with density-gradient centrifugation? Patients randomized to microfluidic sperm preparation had similar cleavage- and blastocyst-stage embryo quality and clinical and ongoing pregnancy rates to those who underwent standard sperm processing for IVF with ICSI. Microfluidic sperm preparation can isolate spermatozoa for clinical use with minimal DNA fragmentation but with unclear impact on clinical outcomes. A prospective randomized controlled trial of 386 patients planning IVF from June 2017 through September 2021 was carried out. One hundred and ninety-two patients were allocated to sperm processing with a microfluidic sperm separation device for ICSI, while 194 patients were allocated to clinical standard density-gradient centrifugation (control) at an academic medical centre. In an intention to treat analysis, there were no differences in high-quality cleavage-stage embryo fraction [66.0 (25.8)% control versus 68.0 (30.3) microfluidic sperm preparation, P = 0.541, absolute difference -2.0, 95% CI (-8.5, 4.5)], or high-quality blastocyst fraction [37.4 (25.4) control versus 37.4 (26.2) microfluidic sperm preparation, P = 0.985, absolute difference -0.6 95% CI (-6, 5.9)] between groups. There were no differences in the clinical pregnancy or ongoing pregnancy rates between groups. The population studied was inclusive and did not attempt to isolate male factor infertility cases or patients with a history of elevated sperm DNA fragmentation. Microfluidic sperm separation performs similarly to density-gradient centrifugation in sperm preparation for IVF in an unselected population. No external funding to declare. M.P.R. is a member of the Clinical Advisory Board for ZyMōt® Fertility, Inc. NCT03085433. 21 March 2017. 16 June 2017.

211 Surgical sperm retrieval (SSR) for management of azoospermia: 1 Year report from a tertiary fertility centre (UK)

Introduction: Infertility affects approximately 1 in 7 heterosexual couples in the UK, with 30% of cases attributed solely to male factor infertility. Surgical Sperm Retrieval (SSR) is a treatment method used to assist with conception for couples with azoospermic men. This audit aimed to retrospectively analyse the profile of male factor infertility patients undergoing SSR in a tertiary care centre over one year period. Methods: Retrospective data was collected for 29 heterosexual couples who visited the fertility clinic at Birmingham Women’s and Children’s Hospital between October 2019 and October 2020.

Male factor infertility trends throughout the last 10years: Report from a tertiary-referral academic andrology centre.

Trends of male factor causes of couples' infertility over time have been poorly investigated. We investigated trends in the causes of pure male factor infertility (MFI) throughout the last 10years in a tertiary-referral academic andrology center. Baseline characteristics at first presentation from a cohort of 1647 consecutive male factor infertility patients belonging to primary infertile couples between 2008 and 2018 have been comprehensively collected over time. Seven major causes of male factor infertility were identified: varicocoele; history of cryptorchidism; hypogonadism (primary and secondary); obstructive azoospermia; genetic abnormalities; other causes (large group including the remnant conditions of known causes); and idiopathic infertility. Rates of different male factor infertility causes over the study period were analyzed. Multivariable logistic regression models tested the likelihood of male factor infertility causes over time. Estimated trends were explored graphically. Of all, varicocoele was found in 615 (37.3%), cryptorchidism in 124 (7.5%), genetic abnormalities in 61 (3.7%), hypogonadism in 165 (10%), obstructive conditions in 55 (3.3%), other causes in 129 (7.8%) patients, and idiopathic infertility in 498 (30.3%) patients, respectively. Over time, a reduction in the proportions of cryptorchidism and varicocoele (all P<0.001) cases was observed, along with an increase in the proportions of hypogonadism, other causes of MFI and idiopathic cases (all P≤0.01). Rates of genetic and obstructive cases remained stable. The observed trends were confirmed at logistic regression models. A decreasing trend in the proportions of varicocoele and cryptorchidism at first presentation was observed over the last 10years; conversely, the proportions of idiopathic cases, hypogonadal patients, and infertile men presenting with other male factor infertility causes significantly increased over the same time frame at a single tertiary-referral academic andrology center.

Endometriosis does not impact live-birth rates in frozen embryo transfers of euploid blastocysts

To determine whether subfertility in patients with endometriosis is due to impaired endometrial receptivity by comparing pregnancy and live-birth outcomes in women with endometriosis versus two control groups without suspected endometrial factors: noninfertile patients who underwent assisted reproduction to test embryos for a single-gene disorder and couples with isolated male factor infertility. Retrospective cohort. Multicenter private practice. All patients aged 24 to 44 years undergoing euploid frozen blastocysts transfer from January 2016 through March2018. None. Live birth, clinical pregnancies, pregnancy losses, and aneuploid rates in preimplantation genetic testing for aneuploidy cycles. The analysis included 459 euploid frozen embryo transfer cycles among 328 unique patients. There were no differences in clinical pregnancy, pregnancy loss, or live-birth rates in patients with endometriosis compared with both control groups. The aneuploidy rates were lowest in the preimplantation genetic testing for monogenic disorders cohort, and the endometriosis patients had aneuploidy rates similar to those of the male factor infertility patients. It is unclear whether endometriosis primarily affects invitro fertilization outcomes via oocyte quality or the endometrium. By controlling for embryo quality using euploid frozen embryo transfer cycles, we found no difference in pregnancy outcomes in patients with endometriosis compared with patients undergoing treatment for male factor infertility and noninfertile patients.

Fertility preservation outcomes in women with gliomas: a retrospective case-control study.

With advances in cancer therapy, reproductive-aged women can look forward to a life post-malignancy. Fortunately, fertility preservation (FP) may provide relief from potential infertility caused by cancer or associated caustic treatments. Outcomes of FP in pre-treatment reproductive-aged women with gliomas have not been previously characterized. Between 2007 and 2018, 10 patients undergoing FP prior to chemotherapy and/or radiation treatment for gliomas were identified at Brigham and Women's Hospital. They were matched 3:1 to male-factor infertility patients by age ± 1year. Patients with gliomas had significantly lower baseline anti-Müllerian hormone levels than male-factor infertility controls (2.37 vs 5.16ng/mL, p = 0.002, log transformed). Despite higher starting (350 vs. 240IU, p = 0.004) and total gonadotropin doses (4270 vs. 2270IU, p < 0.001) over a similar stimulation duration (12.1 vs. 11.1days, p = 0.219), cancer patients had lower peak estradiol levels (1420 vs. 2245pg/mL, p = 0.003). The total number of follicles on the day of trigger (14.1 vs. 15.6, p = 0.284), the number of oocytes retrieved (18.4 vs. 20.5, p = 0.618), and the percentage of mature oocytes (69.9 vs. 73.8%, p = 0.076) were similar between cases and controls. One patient returned for a cryopreserved embryo transfer and delivered a healthy child. Patients undergoing FP prior to chemotherapy and/or radiation for a glioma achieve satisfactory FP outcomes and should be appropriately counseled regarding the opportunity to family-build after treatment.

Seminal Fluid Metabolomic Markers of Oligozoospermic Infertility in Humans

Infertility affects 12–15% of couples worldwide, and male factors are the cause of nearly half of all cases. Studying seminal fluid composition could lead to additional diagnostic accuracy and a better understanding of the pathophysiology of male factor infertility. Metabolomics offers a new opportunity to evaluate biomarkers and better understand pathological mechanisms. The aim of the study was to identify new markers or therapeutic targets to improve outcomes in male factor or idiopathic infertility patients. Semen samples were obtained from 29 men with a normal spermogram test, and from 18 oligozoospermic men. Samples were processed and analyzed by Nuclear Magnetic Resonance spectroscopy and, subsequently, multivariate and univariate statistical analyses. Receiving Operator Curves (ROC) and Spearman correlations were also performed. An Orthogonal Partial Least Square Discriminant Analysis supervised multivariate model was devised to compare the groups. The levels of fructose, myo-inositol, aspartate and choline were altered. Moreover, Spearman Correlation associated fructose, aspartate and myo-inositol with the total amount of spermatozoa, total motile spermatozoa, % of immotility and % of “in situ” spermatozoic motility respectively. NMR-based metabolomics allowed the identification of a specific metabolic fingerprint of the seminal fluids of patients affected by oligozoospermia.

DGC/Zeta as A New Strategy to Improve Clinical Outcome in Male Factor Infertility Patients following Intracytoplasmic Sperm Injection: A Randomized, Single-Blind, Clinical Trial.

Objective The aim of this blind randomised clinical trial study was to assess the clinical efficiency of combined density gradient centrifugation/Zeta (DGC/Zeta) sperm selection procedure compared to conventional DGC in infertile men candidates for intracytoplasmic sperm injection (ICSI). The literature shows that DGC/Zeta is more effective compared to DGC alone in selection of sperms with normal chromatin and improves the clinical outcome of the ICSI procedure. Therefore, this study re-evaluates the efficiency of DGC/Zeta in improving the clinical outcomes of ICSI in an independent clinical setting. Materials and Methods In this randomized, single-blind, clinical trial, a total of 240 couples with male factor infertility and at least one abnormal sperm parameter were informed regarding the study and 220 participated. Based on inclusion and exclusion criteria, 103 and 102 couples were randomly allocated into the DGC/Zeta and DGC groups, respectively. ICSI outcomes were followed and compared between the two groups. Results Although there was no significant difference in fertilization rate (P=0.67) between the DGC/Zeta and DGC groups, mean percentage of good embryo quality (P=0.04), good blastocysts quality (P=0.049), expanded blastocysts (P=0.007), chemical pregnancies (P=0.005) and clinical pregnancies (P=0.007) were significantly higher in the DGC/ Zeta group compared to DGC. In addition, implantation rate was insignificantly higher in DGC/Zeta compared to DGC (P=0.17). ConclusionThis is the second independent study showing combined DGC/Zeta procedure improves ICSI outcomes, especially the pregnancy rate, compared to the classical DGC procedure and this is likely related to the improved quality of sperm selected by the DGC/Zeta procedure (Registration number: IRCT20180628040270N1).

Endometriosis does not impact live birth rates in frozen embryo transfer (FET) of euploid blastocysts

One explanation proposed for subfertility among women with endometriosis is impaired endometrial receptivity. We sought to test this hypothesis by comparing pregnancy and live birth (LB) outcomes in women with endometriosis versus two control groups without suspected endometrial factor: (1) non-infertile patients who underwent ART in order to test embryos for a single gene disorder and (2) couples with isolated male factor infertility. Retrospective Cohort. FETs of PGT-A normal blastocysts performed from January 2016 through March 2018 were included in the analysis. Patients with endometriosis were compared to those with male factor infertility and non-infertile patients using PGT-M for a single gene disorder. Endometriosis was confirmed surgically in 90% of the endometriosis cohort. Patients with multiple infertility diagnoses and those using gestational carriers or donor oocyte were excluded from the analysis. All blastocysts vitrified and warmed for transfer were grade BB or better. Comparisons were made with multigroup chi-square and P<0.05 was considered statistically significant. 472 euploid FET cycles were available for analysis. 59 transfers occurred in patients with endometriosis, 362 transfers in patients with male factor infertility, and 51 transfers in non-infertile patients. There was no difference in patient age in each treatment group and age was not associated with live birth in euploid embryo transfers. An equal number of embryos were transferred in each group. Patients with endometriosis had similar rates of clinical pregnancy (CP) and spontaneous abortion (SAB) when compared to male factor and non-infertile patients. There was no difference in LB in patients with endometriosis (63%) compared to patients with male factor (51%) and non-infertile patients (53%). While the study was only powered to detect a 20% decrease in LB, the 63% LB rate in endometriosis patients did not suggest a negative effect.Tabled 1Endometriosis (n = 59)Male Factor (n = 362)PGT-M (n = 51)P valueNo. Embryos Transferred1.051.091.060.97Positive hCG80%74%80%0.44CP73%65%63%0.43SAB10%14%10%0.63LB63%53%51%0.34 Open table in a new tab Whether endometriosis primarily affects IVF outcomes via oocyte quality or the endometrium is debated. By controlling for embryo quality using euploid FET cycles, we found no difference in pregnancy outcomes in patients with endometriosis compared to those with male factor and non-infertile patients.

Inheritance of epigenetic dysregulation from male factor infertility has a direct impact on reproductive potential

To evaluate the epigenetic consequence on the methylome and subsequent transcriptome in euploid blastocysts of male-factor (MF) infertility patients. Methylome and transcriptome analysis on individual oligoasthenoteratozoospermia (OAT [MF]) blastocysts. Infertility clinic. Clinical data from 128 couples presenting with OAT (MF) and 118 maternal age-matched control (no MF) subjects undergoing infertility treatment from 2010 to 2014, along with 72 surplus cryopreserved blastocysts donated from 33 couples with their informed consents. None. Methyl Maxi-Seq (Zymo Research) was used to determine genome-wide DNA methylation, and small cell number RNA-Seq was used to examine the global transcriptome. Validation experiments were performed with the use of pyrosequencing or quantitative real-time polymerase chain reaction. Statistical analysis used Student t test, analysis of variance in R, Fisher exact test, and pairwise fixed reallocation randomization test where appropriate, with significance at P<.05. Clinical pregnancy rates were similar between OAT (MF) patients and control (no MF) subjects after euploid embryo transfer. However, the miscarriage rate for OAT (MF) patients was significantly higher (14.7% vs. 2.2%; P<.05). Methylome and transcriptome analyses of individual blastocysts revealed significant alterations in 1,111 CpG sites and 469 transcripts, respectively (P<.05). Pathway analysis elucidated genes involved in "regulation of cellular metabolic process" as universally affected. Validation of the genome-wide approaches was performed for SBF1 and SLC6A9 (P<.05). Methylation and transcription aberrations in individual OAT (MF) blastocysts illustrate an epigenetic consequence of MF infertility on embryogenesis, significantly altering key developmental genes and affecting embryonic competence. This epigenetic dysregulation provides an explanation for the reduced reproductive potential in OAT (MF) patients despite euploid blastocyst transfers.

Can intracytoplasmic Morphologically Selected Spermatozoa Injection be Used as First Choice of Treatment for Severe Male Factor Infertility Patients?

Aim:This study was carried out to assess the outcome of the intracytoplasmic morphologically selected sperm injection (IMSI) technique compared with the previous failed intracytoplasmic sperm injection (ICSI) attempts in oligoasthenoteratozoospermia (OAT)/severe OAT (SOAT)/teratozoospermia patients.Setting:Institution-based, in vitro fertilization center.Study Design:It was a nonrandomized prospective study including 57 couples who had previous one or two ICSI failures (failure due to no implantation as embryos were transferred in these cycles and had no pregnancy) due to male factor. There was no case of total fertilization failure. IMSI was carried out in these couples and the results were compared with their previously failed ICSI attempts.Materials and Methods:Real-time selection of sperms was done using IMSI as it allows the assessment of fine nuclear morphology and vacuoles in the sperm head at a high magnification (>6000×) with differential interference contrast optics. Therefore, IMSI was applied in couples having OAT, SOAT or teratozoospermia as male factor and the results were compared with their previous failed ICSI attempts. Statistical analysis was carried out using GraphPad Prism.Results and Conclusion:Data analysis demonstrated significant difference in the fertilization rate between IMSI and previous ICSI attempts of these patients (30.0% vs. 52.0%; P < 0.05). The embryo quality, implantation and pregnancy rates with IMSI were also significantly higher than those of their previous ICSI cycles (32% vs. 56.4%; 30.2% vs. 68.5%; 0.0% vs. 62.4%; P < 0.05). Our conclusion is that the IMSI procedure improved embryo development and the clinical outcomes in the same infertile couples with male infertility and poor embryo development over their previous ICSI attempts and can be taken up as the treatment of choice in cases of severe male factor infertility.

Alterations in the sperm histone-retained epigenome are associated with unexplained male factor infertility and poor blastocyst development in donor oocyte IVF cycles.

Is there a distinct sperm histone-retained epigenetic signature in unexplained male factor infertility patients resulting in compromised blastocyst development? Using only donor oocyte IVF cycles, sperm DNA methylation patterns and miRNA profiles were significantly altered in normozoospermic patients resulting in poor blastocyst development, reflecting a subset of unexplained male factor infertility. Aberrant sperm DNA methylation has been associated with known male factor infertility, particularly noted in oligozoospermic patients. Unexplained male factor infertility remains a significant proportion of in vitro fertilization failures having unknown underlying physiology. Sperm samples (n = 40) and blastocysts (n = 48) were obtained during fertile donor oocyte IVF cycles with normozoospermic parameters, thereby excluding known female and male infertility factors. Samples were divided into two groups based on blastocyst development (Good Group = ≥20% embryos with D5 grade 'AA' blastocysts, and ≥60% embryos of transferable quality on D5 and D6; Poor Group = ≤10% embryos with D5 grade 'AA' blastocysts, and ≤40% embryos of transferable quality on D5 and D6). Samples were obtained from patients undergoing IVF treatments with informed consent and institutional review board approval. The Infinium HumanMethylation450 BeadChip microarray was used to identify histone-retained CpG island genes and genomic regions showing differences in sperm DNA methylation between the Good Group and the Poor Group. Pathway and gene network analysis for the significantly altered genes was performed, and targeted DNA methylation validation was completed for 23 genes and two imprinting control regions. Sperm miRNA profiles were assessed using the TaqMan® Human MicroRNA Array Card, with corresponding blastocyst mRNA gene expression examined by qRT-PCR. Our study is the first to investigate unexplained male factor infertility while significantly eliminating confounding female factors from our sample population by using only young fertile donor oocytes. We identified 1634 CpG sites located at retained histone CpG island regions that had significant sperm DNA methylation differentials between the two embryogenesis groups (P < 0.05). A largely hypermethylated profile was evident in the Good Group, with a small but distinct and statistically significant shift (P < 0.05) observed in the Poor Group. Genes involved in embryonic development were highly represented among histone-retained CpG sites with decreased methylation in the Poor Group (P < 0.05). Ten significantly altered sperm miRNAs (P < 0.05), correlated with altered target gene mRNA expression in the blastocysts from the Poor Group (P < 0.05). Taken together, significantly impacted sperm miRNA and target transcript levels in blastocysts from the Poor Group may contribute alongside aberrant sperm DNA methylation to the compromised blastocyst development observed. Our examination of CpG island regions restricted to retained histones represents only a small part of the sperm epigenome. The results observed are descriptive and further studies are needed to elucidate the functional effects of differential sperm DNA methylation on unexplained male factor infertility and blastocyst development. Slight epigenetic changes in sperm may have a cumulative effect on fertility and embryonic developmental competence. Knowledge of sperm epigenetics and inheritance has important implications for future generations, while providing evidence for potential causes of unexplained male factor infertility. No external funding was used for this study. None of the authors have any competing interest.

Diagnostic application of oxidation-reduction potential assay for measurement of oxidative stress: clinical utility in male factor infertility

The objectives of this study were to: (i) describe a protocol measuring the oxidation-reduction potential (ORP) by MiOXSYS System as an alternative method of seminal oxidative stress (OS) testing; (ii) establish a reference value for static ORP (sORP) to distinguish between controls and male factor infertility patients; (iii) evaluate intra-observer and inter-observer reliability; and (iv) examine association of sORP with sperm parameters predictive of male factor infertility. Elevated levels of sORP were seen in infertile patients (6.22 ± 1.10 mV/106 sperm/ml) compared with controls (1.59 ± 0.29 mV/106 sperm/ml) (P = 0.004). A sORP cut-off value 1.36 mV/106 sperm/ml identified normal semen and abnormal semen quality with a sensitivity 69.6%, specificity 83.1%, positive predictive value 85.3% and negative predictive value 65.9%. The test demonstrated strong intra-observer (CV 8.39%) and inter-observer reliability (correlations >0.97). Higher sORP levels were associated with poor sperm parameters across the fertility status of subjects. Negative correlations were noted with sperm parameters (concentration, total sperm count, motility and morphology) indicating these male infertility parameters are related to OS. In conclusion, the introduction of ORP as a novel clinical test for assessment of OS will help clinicians to better diagnose and manage male factor infertility patients.

Euploid blastocyst transfer is a viable clinical option for male factor infertility with high sperm DNA fragmentation

A significant association has been reported between sperm DNA fragmentation and sperm chromosome abnormalities. The objective of this study was to evaluate the clinical efficacy of euploid frozen blastocyst transfer for male factor (MF) infertility patients with high sperm DNA fragmentation.

Zeta Sperm Selection Improves Pregnancy Rate and Alters Sex Ratio in Male Factor Infertility Patients: A Double-Blind, Randomized Clinical Trial.

Selection of sperm for intra-cytoplasmic sperm injection (ICSI) is usually considered as the ultimate technique to alleviate male-factor infertility. In routine ICSI, selection is based on morphology and viability which does not necessarily preclude the chance injection of DNA-damaged or apoptotic sperm into the oocyte. Sperm with high negative surface electrical charge, named "Zeta potential", are mature and more likely to have intact chromatin. In addition, X-bearing spermatozoa carry more negative charge. Therefore, we aimed to compare the clinical outcomes of Zeta procedure with routine sperm selection in infertile men candidate for ICSI. From a total of 203 ICSI cycles studied, 101 cycles were allocated to density gradient centrifugation (DGC)/Zeta group and the remaining 102 were included in the DGC group in this prospective study. Clinical outcomes were com- pared between the two groups. The ratios of Xand Y bearing sperm were assessed by fluorescence in situ hybridization (FISH) and quantitative polymerase chain reaction (qPCR) methods in 17 independent semen samples. In the present double-blind randomized clinical trial, a significant increase in top quality embryos and pregnancy rate were observed in DGC/Zeta group compared to DGC group. Moreover, sex ratio (XY/XX) at birth significantly was lower in the DGC/Zeta group compared to DGC group despite similar ratio of X/Y bearings sper- matozoa following Zeta selection. Zeta method not only improves the percentage of top embryo quality and pregnancy outcome but also alters the sex ratio compared to the conventional DGC method, despite no significant change in the ratio of Xand Ybearing sperm population (Registration number: IRCT201108047223N1).

High Frequency of Imprinted Methylation Errors in Human Preimplantation Embryos.

Assisted reproductive technologies (ARTs) represent the best chance for infertile couples to conceive, although increased risks for morbidities exist, including imprinting disorders. This increased risk could arise from ARTs disrupting genomic imprints during gametogenesis or preimplantation. The few studies examining ART effects on genomic imprinting primarily assessed poor quality human embryos. Here, we examined day 3 and blastocyst stage, good to high quality, donated human embryos for imprinted SNRPN, KCNQ1OT1 and H19 methylation. Seventy-six percent day 3 embryos and 50% blastocysts exhibited perturbed imprinted methylation, demonstrating that extended culture did not pose greater risk for imprinting errors than short culture. Comparison of embryos with normal and abnormal methylation didn’t reveal any confounding factors. Notably, two embryos from male factor infertility patients using donor sperm harboured aberrant methylation, suggesting errors in these embryos cannot be explained by infertility alone. Overall, these results indicate that ART human preimplantation embryos possess a high frequency of imprinted methylation errors.