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For ejaculated and testicular specimen sources, fertilization patterns, embryo quality, together with implantation are provided for the control and each sperm search time.
Source publication
Introduction:
Severely compromised spermatogenesis typical of men with virtual azoospermia or non-obstructive azoospermia requires an extreme search for spermatozoa. Our goal was to evaluate the usefulness of a meticulous search carried out in ejaculated or surgically retrieved specimens in achieving pre- and post-implantation embryo development....
Similar publications
In males with nonobstructive azoospermia, one of the main histopathologic patterns of the testis is Sertoli cell-only syndrome (SCOS), in which no germ cells are present and only Sertoli cells are contained in the seminiferous tubules. There is not any formal treatment for this pathological condition. However, several studies reported the possibili...
At the beginning of the 21st century there is a decline in quality of reproductive health of men around the world. In structure of sterile marriage the men's factor of infertility is found out in approximately 40-50% of cases. A little studied question of male infertility is the question of quantity and quality of gametes in convoluted seminiferous...
Introduction: Testicular biopsy is important in categorizing patients with Azoospermia and provides useful information and guidelines for further treatment. Histopathological findings of testicular biopsies are of significant importance in making decision for selection of cases for Intracytoplasmic sperm injection (ICSI) in patients with non-obstru...
Citations
... Male infertility accounts for approximately half of the causes of the inability to reproduce among infertile couples [1]. Fortunately, the introduction of ICSI has enhanced the treatment of even the most extreme forms of male factor infertility [2]. Among the different etiologies of male reproductive failure, the most challenging is azoospermia, which accounts for approximately 30% of all cases [3]. ...
... To define azoospermia, a detailed assessment of the patient's medical history is performed, and various evaluations are carried out [6]. In cases of obstructive azoospermia (OA), assessment of the ejaculate does not yield any spermatozoa, whereas in cases of non-obstructive azoospermia (NOA), the ejaculate may inconsistently yield spermatozoa, representing an additional challenge that often requires an extensive search to identify sperm cells [2]. ...
... Once surgical approach is considered, chances of successful sperm retrieval in OA cases is likely to be >99%, while cases with testicular failure can be very unpredictable [2,7]. Furthermore, if spermatozoa are identified in NOA cases, there also lies the ambiguity of whether azoospermia is due to primary testicular failure or depends on secondary environmental factors [8]. ...
Purpose
To identify germline mutations related to azoospermia etiology and reproductive potential of surgically retrieved spermatozoa, and to investigate the feasibility of predicting seminiferous tubule function of nonobstructive azoospermic men by transcriptomic profiling of ejaculates.
Materials and methods
Sperm specimens were obtained from 30 men (38.4 ± 6 years) undergoing epididymal sperm aspiration for obstructive azoospermia (OA, n = 19) acquired by vasectomy, or testicular biopsy for nonobstructive azoospermia (NOA, n = 11). To evaluate for a correlation with azoospermia etiology, DNAseq was performed on surgically retrieved spermatozoa, and cell-free RNAseq on seminal fluid (n = 23) was performed to predict spermatogenesis in the seminiferous tubule.
Results
Overall, surgically retrieved sperm aneuploidy rates were 1.7% and 1.8% among OA and NOA cohorts, respectively. OA men carried housekeeping-related gene mutations, while NOA men displayed mutations on genes involved in crucial spermiogenic functions (AP1S2, AP1G2, APOE). We categorized couples within each cohort according to ICSI clinical outcomes to investigate genetic causes that may affect reproductive potential. All OA-fertile men (n = 9) carried mutations in ZNF749 (sperm production), whereas OA-infertile men (n = 10) harbored mutations in PRB1, which is essential for DNA replication. NOA-fertile men (n = 8) carried mutations in MPIG6B (stem cell lineage differentiation), whereas NOA-infertile individuals (n = 3) harbored mutations in genes involved in spermato/spermio-genesis (ADAM29, SPATA31E1, MAK, POLG, IFT43, ATG9B) and early embryonic development (MBD5, CCAR1, PMEPA1, POLK, REC8, REPIN1, MAPRE3, ARL4C). Transcriptomic assessment of cell-free RNAs in seminal fluid from NOA men allowed the prediction of residual spermatogenic foci.
Conclusions
Sperm genome profiling provides invaluable information on azoospermia etiology and identifies gene-related mechanistic links to reproductive performance. Moreover, RNAseq assessment of seminal fluid from NOA men can help predict sperm retrieval during testicular biopsies.
... ICSI is a widely used technique in assisted reproductive technology (ART), where a single sperm is directly injected into an egg for fertilization [2]. It has definitely revolutionized the treatment of male infertility, particularly in cases of severe oligozoospermia or azoospermia [3]. On the contrary, the rise of technology and growing reliance of clinicians and couples on ARTs allows severe male infertility to be sidestepped through ICSI. ...
... ICSI accounts for up to 80% of all ART cases. It has significantly improved the success rates of ART, especially in cases of male factor infertility, which accounts for 50% of all infertility cases [3]. Nonetheless, it is crucial to juxtapose this recognition with an awareness of the potential overutilization of ICSI in a variety of contexts. ...
... Advanced technologies, such as microfluidic sperm sorting, sperm DNA fragmentation analysis, and sperm proteomics, are emerging, but their implementation in clinical practice is limited by 'andrological ignorance' as well as insufficient trained andrologists critical for proper case-analysis and safe deployment of any emerging technology [15]. The lack of andrological knowledge and advanced technologies in male infertility diagnosis and management contributes to the ICSI paradox by perpetuating the reliance on ICSI as the preferred method of fertilization in severe male factor infertility cases [3]. Therefore, the issue of male infertility frequently does not get the necessary focus and consideration in terms of diagnosis and treatment. ...
The quandary known as the Intracytoplasmic Sperm Injection (ICSI) paradox is found at the juncture of Assisted Reproductive Technology (ART) and 'andrological ignorance'-a term coined to denote the undervalued treatment and comprehension of male infertility. The prevalent use of ICSI as a solution for severe male infertility, despite its potential to propagate genetically defective sperm, consequently posing a threat to progeny health, illuminates this paradox. We posit that the meteoric rise in Industrial Revolution 4.0 (IR 4.0) and Artificial Intelligence (AI) technologies holds the potential for a transformative shift in addressing male infertility, specifically by mitigating the limitations engendered by 'andrological ignorance. ' We advocate for the urgent need to transcend andrological ignorance, envisaging AI as a cornerstone in the precise diagnosis and treatment of the root causes of male infertility. This approach also incorporates the identification of potential genetic defects in descendants, the establishment of knowledge platforms dedicated to male reproductive health, and the optimization of therapeutic outcomes. Our hypothesis suggests that the assimilation of AI could streamline ICSI implementation, leading to an overall enhancement in the realm of male fertility treatments. However, it is essential to conduct further investigations to substantiate the efficacy of AI applications in a clinical setting. This article emphasizes the significance of harnessing AI technologies to optimize patient outcomes in the fast-paced domain of reproductive medicine, thereby fostering the well-being of upcoming generations.
... For Experiment 3, COCs were cultured as outlined above for 28 h then denuded, assessed for MII formation, and imaged as a group. Individual oocytes were inseminated with sperm from one of three sperm donors via ICSI on Day 1 post-retrieval using sperm of suitable quality (Palermo et al., 2014), then cultured in an Embryo Culture Medium (Global Total, Cooper Surgical, Bedminster, NJ) at 37 C in a tri-gas incubator with CO 2 adjusted so that the pH of the bicarbonate-buffered medium was 7.2-7.3, with the O 2 level at 5%. ...
STUDY QUESTION
Can in vitro maturation (IVM) and developmental competence of human oocytes be improved by co-culture with ovarian support cells (OSCs) derived from human-induced pluripotent stem cells (hiPSCs)?
SUMMARY ANSWER
OSC-IVM significantly improves the rates of metaphase II (MII) formation and euploid Day 5 or 6 blastocyst formation, when compared to a commercially available IVM system.
WHAT IS KNOWN ALREADY
IVM has historically shown highly variable performance in maturing oocytes and generating oocytes with strong developmental capacity, while limited studies have shown a positive benefit of primary granulosa cell co-culture for IVM. We recently reported the development of OSCs generated from hiPSCs that recapitulate dynamic ovarian function invitro.
STUDY DESIGN, SIZE, DURATION
The study was designed as a basic science study, using randomized sibling oocyte specimen allocation. Using pilot study data, a prospective sample size of 20 donors or at least 65 oocytes per condition were used for subsequent experiments. A total of 67 oocyte donors were recruited to undergo abbreviated gonadotropin stimulation with or without hCG triggers and retrieved cumulus–oocyte complexes (COCs) were allocated between the OSC-IVM or control conditions (fetal-like OSC (FOSC)-IVM or media-only IVM) in three independent experimental design formats. The total study duration was 1 April 2022 to 1 July 2023.
PARTICIPANTS/MATERIALS, SETTING, METHODS
Oocyte donors between the ages of 19 and 37 years were recruited for retrieval after informed consent, with assessment of anti-Mullerian hormone, antral follicle count, age, BMI and ovarian pathology used for inclusion and exclusion criteria. In experiment 1, 27 oocyte donors were recruited, in experiment 2, 23 oocyte donors were recruited, and in experiment 3, 17 oocyte donors and 3 sperm donors were recruited. The OSC-IVM culture condition was composed of 100 000 OSCs in suspension culture with hCG, recombinant FSH, androstenedione, and doxycycline supplementation. IVM controls lacked OSCs and contained either the same supplementation, FSH and hCG only (a commercial IVM control), or FOSCs with the samesupplementation(Media control). Experiment 1 compared OSC-IVM, FOSC-IVM, and a Media control, while experiments 2and 3 compared OSC-IVM and a commercial IVM control. Primary endpoints in the first two experiments were the MII formation (i.e. maturation) rate and morphological quality assessment. In the third experiment, the fertilization and embryo formation rates were assessed with genetic testing for aneuploidy and epigenetic quality in blastocysts.
MAIN RESULTS AND THE ROLE OF CHANCE
We observed a statistically significant improvement (∼1.5×) in maturation outcomes for oocytes that underwent IVM with OSCs compared to control Media-IVM and FOSC-IVM in experiment 1. More specifically, the OSC-IVM group yielded a MII formation rate of 68% ± 6.83% SEM versus 46% ± 8.51% SEM in the Media control (P = 0.02592, unpaired t-test). FOSC-IVM yielded a 51% ± 9.23% SEM MII formation rate which did not significantly differ from the media control (P = 0.77 unpaired t-test). Additionally, OSC-IVM yielded a statistically significant ∼1.6× higher average MII formation rate at 68% ± 6.74% when compared to 43% ± 7.90% in the commercially available IVM control condition (P = 0.0349, paired t-test) in experiment 2. Oocyte morphological quality between OSC-IVM and the controls did not significantly differ. In experiment 3, OSC-IVM oocytes demonstrated a statistically significant improvement in Day 5 or 6 euploid blastocyst formation per COC compared to the commercial IVM control (25% ± 7.47% vs 11% ± 3.82%, P = 0.0349 logistic regression). Also in experiment 3, the OSC-treated oocytes generated blastocysts with similar global and germline differentially methylated region epigenetic profiles compared commercial IVM controls or blastocysts after either conventional ovarian stimulation.
LARGE SCALE DATA
N/A.
LIMITATIONS, REASONS FOR CAUTION
While the findings of this study are compelling, the cohort size remains limited and was powered on preliminary pilot studies, and the basic research nature of the study limits generalizability compared to randomized control trials. Additionally, use of hCG-triggered cycles results in a heterogenous oocyte cohort, and potential differences in the underlying maturation state of oocytes pre-IVM may limit or bias findings. Further research is needed to clarify and characterize the precise mechanism of action of the OSC-IVM system. Further research is also needed to establish whether these embryos are capable of implantation and further development, a key indication of their clinical utility.
WIDER IMPLICATIONS OF THE FINDINGS
Together, these findings demonstrate a novel approach to IVM with broad applicability to modern ART practice. The controls used in this study are in line with and have produced similar to findings to those in the literature, and the outcome of this study supports findings from previous co-culture studies that found benefits of primary granulosa cells on IVM outcomes. The OSC-IVM system shows promise as a highly flexible IVM approach that can complement a broad range of stimulation styles and patient populations. Particularly for patients who cannot or prefer not to undergo conventional gonadotropin stimulation, OSC-IVM may present a viable path for obtaining developmentally competent, mature oocytes.
STUDY FUNDING/COMPETING INTEREST(s)
A.D.N., A.B.F., A.G., B.P., C.A., C.C.K., F.B., G.R., K.S.P., K.W., M.M., P.C., S.P., and M.-J.F.-G. are shareholders in the for-profit biotechnology company Gameto Inc. P.R.J.F. declares paid consultancy for Gameto Inc. P.C. also declares paid consultancy for the Scientific Advisory Board for Gameto Inc. D.H.M. has received consulting services from Granata Bio, Sanford Fertility and Reproductive Medicine, Gameto, and Buffalo IVF, and travel support from the Upper Egypt Assisted Reproduction Society. C.C.K., S.P., M.M., A.G., B.P., K.S.P., G.R., and A.D.N. are listed on a patent covering the use of OSCs for IVM: U.S. Provisional Patent Application No. 63/492,210. Additionally, C.C.K. and K.W. are listed on three patents covering the use of OSCs for IVM: U.S. Patent Application No. 17/846,725, U.S Patent Application No. 17/846,845, and International Patent Application No.: PCT/US2023/026012. C.C.K., M.P.S., and P.C. additionally are listed on three patents for the transcription factor-directed production of granulosa-like cells from stem cells: International Patent Application No.: PCT/US2023/065140, U.S. Provisional Application No. 63/326,640, and U.S. Provisional Application No. 63/444,108. The remaining authors have no conflicts of interest to declare.
... Megjithatë, te një numër i meshkujve infertilë të trajtuar me prednisolone për më shumë se tre muaj, është vërejtur një rritje e shkallës së shtatzanisë, kundrejt placebo grupit. Dhënia e agjentëve imunosupresorë (azathioprine apo cyclophosphamide i.v) është propozuar si trajtim sistematik dhe dhënia intravenoze e 91 imunoglobulinave. TRA mbetet një opcion i artë për të gjithë ata që dëshirojnë të arrijnë një shtatzani të sigurtë. ...
Kurorëzimi e një martese nuk konsiston në bërjen e dasmës, por në momentin kur çifti dashurinë e tyre e finalizon me sigurimin e pasardhësve, një process që po bëhet gjithnjë e më i vështirë për shkak të uljes së shkallës së fertilizimit si tek meshkujt ashtu edhe tek femrat. Të dhënat e dala nga studimet më të fundit janë të frikshme, duke parashikuar një prekje të vijës së limitit minimal të numrit të nevojshëm të spermatozoideve për të fertilizuar qelizën vezë, dhe kjo jo shumë larg, por në 30 vitet e ardhshme, duke vështirësuar tejmase procesin e pasjes së pasardhësve dhe duke i dhënë një theks të veçantë dhe rëndësi metodave të fertilizimit in vitro.
Duke marrë parasysh nevojat që mjekët e së nesërmes do të kenë lidhur me këtë fushë, është paraparë një përgatitje shkollore e tyre gjatë studimeve të mjekësisë duke vendosur lëndë të këtilla në formatin e lëndëve zgjedhore, në mënyrë që ballafaqimi dhe kërkesat që besoj se do të rriten shpejtë për fushën e andrologjisë, të nxjerrë në “fushëbetejë” profesionistë shëndetësore të gatshëm për ta mbajtur të gjallë specien më të rëndësishme të ekosistemit – njeriun.
Ky material është paraparë që të japë bazat e problematikës dhe mënyrën e qasjes bashkëkohore të kësaj problematike. Jam i vetëdijshëm që ky material i paraparë për këtë vit, nuk do të mund të plotësojë të gjitha nevojat dhe kërkesat, prandaj sugjerimet dhe kritikat lidhur me materialin janë më se të mirëseardhura, në mënyrë që të vijë deri te një infomracion sa më sublim dhe kuptimplotë për të gjithë profesionistët por edhe për ata të cilët janë epiqendra e këtij hulumtimi, meshkujt infertilë, baballarët e së nesërmes!
... still not available for both, dogs (23,(79)(80)(81)(82)(83)(84) and men (24,26,(85)(86)(87)(88). Although significantly upregulated PTGS2 expression points to the importance of inflammation as a key process in canine CAO (89), it seems unlikely that selective Cox-2 inhibitors (alone) are suitable to protect fertility indicating the need for other therapeutic options such as assisted reproductive techniques (24,90,91). Stem cell-based therapeutic options (36,42,92) to (re-) initialize spermatogenesis (37,38,40,47,(93)(94)(95)(96)(97)(98) might be a future treatment for CAO-affected dogs, too. ...
Chronic asymptomatic idiopathic orchitis (CAO) is an important but neglected cause of acquired infertility due to non-obstructive azoospermia (NOA) in male dogs. The similarity of the pathophysiology in infertile dogs and men supports the dog's suitability as a possible animal model for studying human diseases causing disruption of spermatogenesis and evaluating the role of spermatogonial stem cells (SSCs) as a new therapeutic approach to restore or recover fertility in cases of CAO. To investigate the survival of resilient stem cells, the expression of the protein gene product (PGP9.5), deleted in azoospermia like (DAZL), foxo transcription factor 1 (FOXO1) and tyrosine-kinase receptor (C-Kit) were evaluated in healthy and CAO-affected canine testes. Our data confirmed the presence of all investigated germ cell markers at mRNA and protein levels. In addition, we postulate a specific expression pattern of FOXO1 and C-Kit in undifferentiated and differentiating spermatogonia, respectively, whereas DAZL and PGP9.5 expressions were confirmed in the entire spermatogonial population. Furthermore, this is the first study revealing a significant reduction of PGP9.5, DAZL, and FOXO1 in CAO at protein and/or gene expression level indicating a severe disruption of spermatogenesis. This means that chronic asymptomatic inflammatory changes in CAO testis are accompanied by a significant loss of SSCs. Notwithstanding, our data confirm the survival of putative stem cells with the potential of self-renewal and differentiation and lay the groundwork for further research into stem cell-based therapeutic options to reinitialize spermatogenesis in canine CAO-affected patients.
... 5,6 Controversy exists over the use of ejaculate versus testicular sperm for ICSI in cryptozoospermic patients. [5][6][7][8][9] Ejaculate sperm is thought to be more mature than testicular sperm. 10 Nevertheless, there are inherent concerns with the use of ejaculate sperm; the repeated centrifugations needed to identify viable sperm may increase the production of reactive oxidative species affecting the quality of the sperm. ...
... On the other hand, TESE has shown to have debatable benefits over ejaculate sperm, [5][6][7][8][9] and it carries risks of surgical complications and long-term adverse effects including hypoandrogenism. 15,16 Additionally, previous studies have described a correlation between testicular extracted sperm and spermatic aneuploidy in patients with non-obstructive azoospermia. ...
Objective The aim of this study is to evaluate the rate of embryonic euploidy in blastocysts derived from testicular versus ejaculated sperm in cryptozoospermic patients. Design Retrospective cohort analysis. Material and methods The study included couples who suffer from Cryptozoospermia and underwent an autologous in vitro fertilization (IVF) with preimplantation genetic testing (PGT-A) cycle(s) from 2014 to 2019. Only cases where oocyte insemination was conducted with intra-cytoplasmic sperm injection (ICSI) were evaluated. Cohorts were separated based on the source of sperm (Ejaculated vs. Testicular (TESE)). Demographic and clinical embryology parameters were compared among cohorts. Student’s t-test, Wilcoxon’ rank test, chi-square test, and multivariate logistic regression fitted with a GEE model were used for data analysis. Results A total of 573 blastocysts derived from 87 IVF/PGT-A cases were included in the study. 74 cases (n= 474 embryos) utilized ejaculated sperm and 13 cases (n= 99 embryos) utilized testicular sperm. No significant differences were found in demographic and stimulation parameters among cohorts. (Table 1) No differences among the ejaculated and testicular cohorts were found in fertilization rate (63.2%; 61.1%, p=0.32); blastulation rate (64.5%; 66.6%, p=0.69); and rate of embryo euploidy (49.7%; 52.1%, p=0.76) respectively. No differences were found in rate of cycle cancellation due to unavailable embryos for TE biopsy (18.9% vs 7.6%, p=0.32). Conclusions There is no genomic advantage to surgical sperm retrieval in cryptozoospermic patients.
... Individual oocytes in each condition were injected with sperm via intracytoplasmic sperm injection (ICSI) on day 1 post-retrieval. 43 After ICSI, the oocytes were cultured in a medium designed for embryo culture (Global Total, Cooper Surgical, Bedminster, NJ) at 37 o C in a tri-gas incubator with CO 2 adjusted so that the pH of the bicarbonate-buffered medium was 7.2-7.3 and with the O 2 level maintained at 5%. 12 to 16 hours post-ICSI, fertilization was assessed and zygotes were micrographed, and oocytes with two pronuclei were cultured until day 3. Cleaved embryos were micrographed, underwent laser-assisted zona perforation and were allowed to develop until the blastocyst stage. ...
Assisted reproductive technologies (ART) have significantly impacted fertility treatment worldwide through innovations such as in vitro fertilization (IVF) and in vitro maturation (IVM). IVM holds promise as a technology for fertility treatment in women who cannot or do not wish to undergo conventional controlled ovarian stimulation (COS). However, IVM has historically shown highly variable performance in maturing oocytes and generating oocytes with strong developmental capacity. Furthermore, recently reported novel IVM approaches are limited to use in cycles lacking human chorionic gonadotropin (hCG) triggers, which is not standard practice in fertility treatment. We recently reported the development of ovarian support cells (OSCs) generated from human induced pluripotent stem cells (hiPSCs) that recapitulate dynamic ovarian function in vitro . Here we investigate the potential of the se OSCs in an IVM co-culture system to improve the maturation of human cumulus-enclosed immature oocytes retrieved from abbreviated gonadotropin stimulated cycles. We reveal that OSC-IVM significantly improves maturation rates compared to existing IVM systems. Most importantly, we demonstrate that OSC-assisted IVM oocytes are capable of significantly improving euploid blastocyst formation and yielding blastocysts with normal global and germline differential methylation region methylation profiles, a key marker of their clinical utility. Together, these findings demonstrate a novel approach to IVM with broad applicability to modern ART practice.
Structured Abstract
Objective
To determine if in vitro maturation (IVM) of human oocytes can be improved by co-culture with ovarian support cells (OSCs) derived from human induced pluripotent stem cells (hiPSCs).
Design
Three independent experiments were performed in which oocyte donors were recruited to undergo abbreviated gonadotropin stimulation and retrieved cumulus oocyte complexes (COCs) were randomly allocated between the OSC-IVM and control IVM conditions.
Subjects
Across the three experiments, a total of 67 oocyte donors aged 19 to 37 years were recruited for retrieval using informed consent. Anti-mullerian hormone (AMH) value, antral follicle count (AFC), age, BMI, and ovarian pathology were used for inclusion and exclusion criteria.
Intervention and Control
The OSC-IVM culture condition was composed of 100,000 OSCs in suspension culture supplemented with human chorionic gonadotropin (hCG), recombinant follicle stimulating hormone (rFSH), androstenedione and doxycycline. IVM controls comprised commercially-available IVM media without OSCs and contained either the same supplementation as above (media-matched control), or FSH and hCG only (IVM media control). In one experiment, an additional control using fetal ovarian somatic cells (FOSCs) was used with the same cell number and media conditions as in the OSC-IVM.
Main Outcome Measures
Primary endpoints consisted of metaphase II (MII) formation rate and oocyte morphological quality assessment. A limited cohort of oocytes were utilized for secondary endpoints, consisting of fertilization and blastocyst formation rates with preimplantation genetic testing for aneuploidy (PGT-A) and embryo epigenetic analysis.
Results
OSC-IVM resulted in a statistically significant improvement in MII formation rate compared to the media-matched control, a commercially available IVM media control, and the FOSC-IVM control. Oocyte morphological quality between OSC-IVM and controls did not significantly differ. OSC-IVM displayed a trend towards improved fertilization, cleavage, and blastocyst formation. OSC-IVM showed statistically significant improvement in euploid day 5 or 6 blastocyst formation compared to the commercially available IVM media control. OSC-IVM embryos displayed similar epigenetic global and germline loci profiles compared to conventional stimulation and IVM embryos.
Conclusion
The novel OSC-IVM platform is an effective tool for maturation of human oocytes obtained from abbreviated gonadotropin stimulation cycles, supporting/inducing robust euploid blastocyst formation. OSC-IVM shows broad utility with different stimulation regimens, including hCG triggered and untriggered oocyte retrieval cycles, making it a highly useful tool for modern fertility treatment.
... Nowadays, it is feasible to treat most forms of male factor infertility, particularly in cases with a very low concentration or impaired characteristics of the spermatozoon. Indeed, even those men with severely dysmorphic gametes, such as the globozoospermic, can be successfully treated by ICSI (1), and even scarce sperm cells retrieved from the seminiferous tubules of nonobstructive azoospermic individuals can be used to successfully inseminate an oocyte (2). ...
Objective
To assess the role of evaluating Sperm Chromatin Fragmentation (SCF) as a tool to guide treatment in couples who achieved unexpectedly poor clinical outcomes following ICSI.
Design
We identified couples with an unexpectedly suboptimal clinical outcome following Intracytoplasmic Sperm Injection (ICSI), who were then screened for SCF. Consequently, the same couples were counselled to undergo a subsequent ICSI cycle using either ejaculates processed by Microfluidic Sperm Selection (MFSS) or spermatozoa retrieved from the testis, and clinical outcomes were compared. To confirm the sole effect of a compromised male gamete, we compared ICSI outcome in cycles where male gametes with abnormal SCF were used to inseminate autologous and donor oocytes. Finally, to eliminate an eventual confounding female factor component, we compared clinical outcome of ICSI cycles utilizing sibling donor oocytes injected with spermatozoa with normal or abnormal SCF.
Setting
Academic Reproductive Medicine Center point of care.
Patient(s)
The patient population consisted of a total of 76 couples with reproductively healthy and relatively young female partners, and male partners with compromised semen parameters, but suitable for ICSI. In a sub analysis, we identified 67 couples with an abnormal SCF who underwent ICSI cycle(s) with donor oocytes. Furthermore, we identified 29 couples, 12 with normal SCF and 17 with abnormal uncorrected SCF, and 7 couples with corrected abnormal SCF spermatozoa verses a control, who used sibling donor oocytes for their ICSI cycle(s).
Interventions
For couples who resulted in surprisingly low clinical outcomes following ICSI, despite semen parameters adequate for ICSI and a normal female infertility evaluation, a SCF assessment was performed on the semen specimen utilizing the TUNEL assay. The couples then underwent a subsequent ICSI cycle with spermatozoa processed by MFSS or surgically retrieved. Moreover, cycles utilizing donor oocytes were used to confirm the sole contribution of the male gamete.
Main Outcome Measure(s)
Clinical outcome such as fertilization, embryo implantation, clinical pregnancy, delivery, and pregnancy loss rates were compared between history and treatment cycle(s) using ejaculated spermatozoa selected by MFSS or from a testicular biopsy, taking into consideration level of SCF. In a sub-analysis, we reported the clinical outcome of 67 patients who used donor oocytes and compared to cycles where their own oocytes were used. Furthermore, we compared ICSI clinical outcomes between cycles using sibling donor oocytes injected with low versus high SCF without or with sperm intervention aimed at correcting the final SCF.
Result
(s): In a total of 168 cycles, 76 couples had in a prior cycle with a 67.1% fertilization, ana a clinical pregnancy and pregnancy loss rate of 16.6% and 52.3%, respectively. Following testing for SCF, DNA fragmentation rate was 21.6%. This led to a subsequent ICSI cycle with MFSS or TESE, resulting in a clinical pregnancy of 39.2% (P<0.01), and delivery of 37.3% (P<0.001). The embryo implantation rose to 23.5% (P<0.001). while the pregnancy loss decreased to 5% in the treatment cycle (P<0.01). This was particularly significant in the moderate SCF group, reaching an embryo implantation of 24.3% (P<0.001), clinical pregnancy of 40.4% (P<0.01), and a delivery rate of 36.2% (P<0.001) reducing the pregnancy loss to 10.5% in post sperm treatment cycles (P<0.01).
In 67 patients with high SCF who utilized donor oocytes a significantly higher fertilization of 78.1% (P<0.00001) and embryo implantation of 29.1% (P<0.0001) was reported, as compared to couples also with an elevated SCF who utilized their own. Interestingly, the clinical pregnancy and delivery rates only increased slightly from 28.0% to 36.1% and from 23.7% to 29.2%, respectively.
To further control for a female factor, we observed couples that shared sibling donor oocytes, 17 with a normal SCF and 12 with an abnormal (uncorrected) SCF. Interestingly, the abnormal SCF group had an impaired fertilization (69.3%, P<0.001), embryo implantation (15.0%, P<0.05), and delivery (15.4%, P<0.05) rate.
For an additional 15 couples who split their donor oocytes, 8 couples had a normal SCF, and while 7 couples originally had abnormal SCF, 4 used microfluidic processing, 2 used testicular spermatozoa and 1 used donor spermatozoa, resulting in comparable clinical outcomes to the normal SCF group.
Conclusion
A superimposed male factor component may explain the disappointing ICSI outcome in some couples despite reproductively healthy female partners. Therefore, it may be useful to screen couples for SCF to guide treatment options and maximize chances of a successful pregnancy. The improved but suboptimal pregnancy and delivery outcomes observed in couples using donor oocytes confirmed the exclusive detrimental role that the male gamete exerted on embryo development despite the presence of putative oocyte repair mechanisms.
... 4,5 With ICSI, fertilization is feasible even when only a few spermatozoa can be identified. 6 Given its versatility and success, ICSI has become the most used method of fertilization in assisted reproductive technology (ART) and represents a reliable intervention to enhance fertility. 7 Despite its widespread application, there are some hurdles in achieving uniform success with ART. ...
The inability to conceive due to male infertility is a complex issue with a wide variety of etiologies. Sperm DNA damage can be both a barrier to natural pregnancy and successful assisted reproductive technology (ART). The aim of this narrative review was to describe and highlight the effects of sperm DNA fragmentation and the most recent data on various treatment strategies to decrease sperm DNA damage. Finally, we proposed a management algorithm for couples undergoing ART with increased sperm DNA fragmentation.
... At present, intracytoplasmic sperm injection (ICSI) is widely used in assisted reproductive technology (ART). The aim of ART is to achieve fertilization by directly injecting the sperm into the oocyte by passing the many biological barriers in the process [7]. Continuous improvement in ART has allowed severe infertility cases to be successful, even when recurrent fertilization failures occur after conventional in vitro fertilization (IVF). ...
... At present, IVF and ICSI are reproductive technologies that are widely used to treat infertility related to reproductive endocrinology, genetic disorders, oocyte donation, and surrogacy. The aim of ART is to attain a successful pregnancy, and during this process, most biological barriers are bypassed, especially when ICSI is applied, because a morphologically normal spermatozoon is directly injected into a mature oocyte [7]. Since the early 1990s, when the first pregnancy using ICSI was reported [88], almost any type of spermatozoa were used to fertilize an oocyte. ...
... Numerous hypotheses have been offered explaining how the sperm activates the oocyte by increasing In has been associated, via immunohistochemistry, the level of PAWP and sperm quality and fertilizing ability It has been proposed that when the PPXY region of PAWP is blocked, the oocyte activation failed [7,[58][59][60] intracellular Ca + 2 oscillations. Many factors and molecular pathways have been studied to determine pronuclear formation, for which the oocyte machinery modifies the sperm chromatin structure after fertilization. ...
The fertilizing spermatozoon is a highly specialized cell that selects from millions along the female tract until the oocyte. The paternal components influence the oocyte activation during fertilization and are fundamental for normal embryo development; however, the sperm-oocyte interplay is in a continuous debate. This review aims to analyze the available scientific information related to the role of the male gamete in the oocyte activation during fertilization, the process of the interaction of sperm factors with oocyte machinery, and the implications of any alterations in this interplay, as well as the advances and limitations of the reproductive techniques and diagnostic tests. At present, both PLCζ and PAWP are the main candidates as oocyte activated factors during fertilization. While PLCζ mechanism is via IP 3 , how PAWP activates the oocyte still no clear, and these findings are important to study and treat fertilization failure due to oocyte activation, especially when one of the causes is the deficiency of PLCζ in the sperm. However, no diagnostic test has been developed to establish the amount of PLCζ, the protocol to treat this type of pathologies is broad, including treatment with ionophores, sperm selection improvement, and microinjection with PLCζ protein or RNA.
