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This study aims to compare the embryological and clinical parameters of intracytoplasmic sperm injection (ICSI) cycles using testicular versus ejaculated sperm in male patients with elevated sperm DNA fragmentation (SDF). A total of 73 ICSI cycles were examined in couples where the male partner exhibited high levels of SDF. ICSI was performed using either ejaculated or testicular sperm. The primary outcomes were rates of blastocyst formation, high-quality embryo development, and clinical pregnancy. The DNA fragmentation index (DFI) for testicular sperm (16.81 ± 17.51) was significantly lower than that of ejaculated sperm (56.96 ± 17.56). While the blastocyst formation rate was significantly higher in the testicular sperm group compared to the ejaculated sperm group, no statistically significant differences were noted in fertilization rate (72.15% vs. 77.23%), rate of high-quality embryo formation (47.17% vs. 46.53%), clinical pregnancy (50% vs. 56.52%), Cumulative pregnancy (70.2% vs. 55.6%), or live birth rate (43.75% vs.43.48%). Testicular spermatozoa have no additional advantage over ejaculated spermatozoa except for blastocyst quality in patients with high SDF, the use of testicular spermatozoa for the first ICSI cycle in male infertility patients with high SDF should be undertaken after much consideration at present. [ABSTRACT FROM AUTHOR]

Purpose: Optimal sperm DNA integrity is essential for fertilization and embryo health. Research indicates that testicular sperm (TS), obtained via TESA or TESE, typically show lower sperm DNA fragmentation (SDF) than ejaculated sperm after standard abstinence. Shortening abstinence to less than 2 days might reduce SDF, offering a less invasive and more cost-effective alternative to surgical sperm retrieval. Yet, no studies have directly compared the efficacy of shorter abstinence against TS extraction for lowering SDF. Our meta-analysis aims to address this gap by comparing SDF levels in TS to those in ejaculated sperm after a short abstinence period. Methods: Meta-analysis of 16 randomized controlled and prospective observational studies included 4 on TS and 12 on short abstinence ejaculation. The meta-analysis followed MOOSE guidelines, scrutinizing databases including Cochrane Library, Web of Science, Embase, MEDLINE(R), and PUMBED up to November 16, 2023. The analysis was conducted using RevMan. The observational studies' methodological quality was assessed using the Newcastle–Ottawa Scale, and the overall evidence quality was evaluated following the GRADE criteria. To compare short ejaculation duration and TS (are not directly compared in the literature) for SDF levels, we analyzed relevant data from studies of each method. We adjusted the participant numbers in the TS group by 1/3 and included each TS study three times, to perform a comparison against the short duration studies which were in a ratio of 1:3. This approach maintained an unaltered cumulative subject count for the meta-analysis of TS studies. Results: A total of 641 patients were included, comprising 120 and 521 patients with SDF measurements following TS and ejaculation after a short abstinence period, respectively. The studies had varied inclusion criteria, with not all patients having an initial elevated SDF. Some studies had incomplete details on age and other demographics. However, the mean ± SD age of 93 TS patients was 38.15 ± 5.48 years vs. 37.7 ± 6.0 years of 444 short abstinence patients, demonstrating no significant difference (P = 0.544). Short abstinence durations ranged from 1 to 48 h. Diverse DNA fragmentation tests were used: TUNEL assay in three testicular sperm studies, SCD assay in one, and in the short abstinence group, four used TUNEL and six used SCD assays, along with one each using SCSA and Halosperm. The mean ± SD SDF was lower in the TS group than in the short abstinence group (mean difference − 9.48, 95%CI − 12.45 to − 6.52, P < 0.001, I2 = 85%). Sensitivity analysis revealed that no single study significantly influenced the results. Employing the GRADE criteria, the initial assessment categorized the overall quality of evidence as low due to the observational nature of the acquired data. All studies were of medium to high quality. Conclusion: This study suggests testicular sperm may be better than ejaculated sperm for improving SDF in infertility cases. Direct comparisons are needed, before deeming short abstinence less effective. Future research should directly compare reproductive outcomes using both methods. [ABSTRACT FROM AUTHOR]

STUDY QUESTION Which independent factors influence ICSI outcomes in patients with complete azoospermia factor c (AZFc) microdeletions? SUMMARY ANSWER 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. WHAT IS KNOWN ALREADY 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. STUDY DESIGN, SIZE, DURATION 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. PARTICIPANTS/MATERIALS, SETTING, METHODS 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. MAIN RESULTS AND THE ROLE OF CHANCE 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. LIMITATIONS, REASONS FOR CAUTION 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. WIDER IMPLICATIONS OF THE FINDINGS 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. STUDY FUNDING/COMPETING INTEREST(S) 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. TRIAL REGISTRATION NUMBER N/A. [ABSTRACT FROM AUTHOR]

Background: Non‐obstructive azoospermia (NOA) diagnosis poses challenges for couples seeking parenthood. Microdissection testicular sperm extraction (MD‐TESE) excels in retrieving testicular sperm cells for NOA cases. However, limited live birth data in Australian NOA patients hinders accurate counselling. Aims: This study aimed to determine the likelihood of infertile couples with a male partner diagnosed with NOA conceiving biological children using MD‐TESE / intracytoplasmic sperm injection (ICSI). Materials and methods: A retrospective cohort study included 108 NOA men treated at a public fertility unit and a private fertility centre (May 2009–May 2022). Primary outcome: live birth rate (LBR); secondary outcomes: sperm retrieval rate, pregnancy rate, and neonatal outcomes. Results: Among 108 patients undergoing MD‐TESE, the positive sperm retrieval rate (PSRR) was 64.8% (70/108). Histology best predicted sperm retrieval success, with hypo‐spermatogenesis yielding a 94.1% PSRR. Age, testicular volume, and hormonal parameters had no significant impact. Mean male age: 35.4 years; mean partner age: 32.7 years. Fertilisation rate: 50.7%. LBR per initiated cycle: 58.7% (37/63); per embryo transfer: 63.8% (37/58); per initially diagnosed NOA man: 34.3% (37/108). Cumulative LBR: 74.1% (43/58); twin rate: 10.8% (4/37). No neonatal deaths or defects were observed among 47 live offspring. Conclusion: This study provides valuable data for counselling NOA couples on the probability of conceiving biological offspring. MD‐TESE and ICSI yielded favourable PSRR (64.8%) and LBR (63.8%). However, couples should be aware that once NOA is confirmed, the chance of taking home a baby is 34%. [ABSTRACT FROM AUTHOR]

Intracytoplasmic sperm injection (ICSI) is clinically used to treat obstructive/nonobstructive azoospermia. This study compared the efficacy of ICSI with cauda epididymal and testicular sperm in Wistar (WI) and Brown- Norway (BN) rats. The transfer of ICSI oocytes with cryopreserved epididymal and testicular WI sperm resulted in offspring production of 26.2% and 3.7%-4.7%, respectively (P < 0.05). Treatments for artificial oocyte activation (AOA) and acrosome removal improved pronuclear formation in BN-ICSI oocytes; however, only AOA treatment was effective in producing offspring (3.7%-6.5%). In the case of ICSI with testicular sperm (TESE-ICSI), one offspring (0.6%) was derived from the BN-TESE-ICSI oocytes. The application of AOA or a hypo-osmotic sperm suspension did not improve the production of TESE-ICSI offspring. Thus, outbred WI rat offspring can be produced by using ICSI and less efficiently by using TESE-ICSI. Challenges in producing offspring by using ICSI/TESE-ICSI in inbred BN strain require further investigation. [ABSTRACT FROM AUTHOR]

Introduction: Although the effectiveness of pentoxifylline (PF) as a selective inhibitor of phosphodiesterase to enhance sperm motility through increasing cyclic nucleotide in cases of absolute asthenozoospermia has been demonstrated for ICSI, data related to babies born from the PF-ICSI are still severely lacking. Concerns have been raised regarding the potential embryotoxicity of PF due to the controversial results obtained from the analysis of this compound on animal embryo development. This study aimed to determine whether the application of PF to trigger frozen-thawed TESA (testicular sperm aspiration) spermatozoa increases the risk of adverse obstetric and neonatal outcomes compared with non-PF frozen-thawed TESA ICSI and conventional ICSI using fresh ejaculation. Materials and methods: A total of 5438 patients were analyzed in this study, including 240 patients underwent PF-TESA ICSI (ICSI using PF triggered frozenthawed testicular spermatozoa), 101 patients underwent non-PF TESA ICSI (ICSI using frozen-thawed testicular spermatozoa) and 5097 patients underwent conventional ICSI using fresh ejaculation. Propensity score matching was executed to control the various characteristics of patients. Results: No significant differences in pregnancy outcomes were observed among the three groups (PF-TESA ICSI, non-PF TESA ICSI and conventional ICSI), including biochemical pregnancy, clinical pregnancy, implantation, miscarriage, ectopic pregnancy, multiple pregnancy, and live birth, following propensity score matching. Additionally, neonatal outcomes were found to be similar among the three groups, with no statistical differences observed in the birth defect, birth weight, gestational age, preterm birth, and earlyneonatal death. Discussion and conclusion: PF-ICSI may be an alternative treatment in patients using frozen-thawed testicular spermatozoa, resulting in comparable pregnancy and neonatal outcomes. [ABSTRACT FROM AUTHOR]

Fish are ectotherms and many have an external reproductive mode. An environmental factor which triggers fish reproductive activity in fish is water temperature. However, climate change is causing increasingly frequent events in which the water temperature varies rapidly; as a result, both in hatchery and in natural conditions, fish sperm are exposed to varying environmental temperatures during their journey toward the egg. This study was based on two experiments: The first experiment was designed to determine how storage at 4 °C for four days affected the sperm functions of Atlantic salmon (Salmo salar) sperm collected by either abdominal massage (stripping/Pure) or testicular dissection (testicular macerate/Macerated). Further, computer-assisted semen analysis (CASA) was used to compare sperm velocity parameters (VCL, VSL, and VAP) and progressivity (STR, LIN, and WOB) after motility activation at different temperatures (8 and 16 °C) of sperm collected by both methods (Pure vs Macerated). The results show that spermatozoa from Macerated samples maintained a higher sperm function when stored at 4 °C for 4 days compared to Pure sperm samples. In the second experiment, CASA determined that all parameters for sperm velocity (VCL, VSL, and VAP) and progressivity (STR (50%/55%), LIN (25%–32%), and WOB (51%–57%) were affected by activation temperature (P < 0.05) and that the motility patterns after activation at 16 °C (P < 0.05), specifically the LIN or STR swimming trajectories of the sperm differed between the two groups. In conclusion, the sperm quality of testicular Macerate was superior to that of Pure sperm abdominal mass, based on the higher quality of various sperm functions during short-term storage. Moreover, there was a significant effect of the temperature of the activation medium on sperm speed and progressivity (motility pattern) in the collected samples of testicular macerate. The sensitivity of Salmo salar spermatozoa to elevated temperature varies markedly between collection methods (Pure and Macerated). • The collection method and temperature motility activation medium affect the physiology of spermatozoa in Salmo salar. • Salmo salar spermatozoa collected by testicular dissection had enhanced cold tolerance during in vitro storage. • Sperm collected by testicular dissection were more sensitive to thermal changes at the time of motility activation. • The sperm motility pattern was affected by the method of sperm collection and the temperature of the activation medium. [ABSTRACT FROM AUTHOR]

Background: Single sperm cryopreservation (SSC) is a specific technique especially used in individuals with small numbers of sperm who suffered from non-obstructive azoospermia (NOA). Testicular specimens possess poor motility and low population of viable spermatozoa. Therefore, sperm selection methods such as applying pentoxifylline (PTX) may improve motility in these cases. The main aim of this study was to evaluate the protective effects of PTX on testicular spermatozoa before and after performing SSC. Methods: Thirty testicular samples were obtained from men with azoospermia. This study was conducted in two phases. Phase 1 evaluated the effect of PTX for sperm selection before SSC. Twenty testicular samples were divided to two experimental groups: SSC without (I) and with PTX treatment (II). For PTX treatment spermatozoa were incubated with PTX at 37°C for 30 min and only motile spermatozoa were selected for SSC. In phase 2, ten testicular samples were cryopreserved with SSC and warming procedure was carried out in droplet with and without PTX. Motility and viability rates, morphology by motile sperm organelle morphology examination (MSOME), DNA fragmentation by sperm chromatin dispersion test (SCD) and mitochondrial membrane potential (MMP) were evaluated. Results: In phase 1, post warm motility rate was higher in PTX exposed group compared to the unexposed group (25.6 ± 8.13 vs. 0.85 ± 2.1) (p > 0.00). Recovery rate, viability and morphology were not significantly different between groups. DNA integrity and MMP were also similar between both groups. In phase 2 although motility increased in PTX group compared to without PTX group (29.30 ± 12.73 vs. 1.90 ± 2.64) (p > 0.00), the viability rate was not different (70.40 ± 12.12 vs. 65.30 ± 11.87). All above mentioned parameters were similar between the two SSC groups. Conclusions: Supplementation of testicular spermatozoa with PTX before cryopreservation increases motility and did not have adverse effects on viability, morphology, DNA integrity and MMP. PTX could be used as sperm selection method before single sperm cryopreservation, but PTX could not maintain motile the most of viable testicular sperms.

Objective To investigate the efficacy, safety and the influence on pregnancy outcome of L-carnitine before intracytoplasmic sperm injection (ICSI) from different sources of sperm in patients with obstructive azoospermia (OA). Methods A total of 141 patients with OA were treated with L-carnitine for three months, and sperms were obtained by testicular sperm aspiration (TESA) and percutaneous epididymal sperm aspiration (PESA) respectively. According to the source of sperm, they were divided into the two groups: the TESA group (n=78) and the PESA group (n=63). The general clinical data, sperm quality, embryonic development and clinical outcome of the two groups were compared. Results In the TESA/PESA group, sperm DFI and sperm spontaneous acrosome reaction rate were significantly lower than those before treatment, and sperm acrosome integrity rate was significantly higher than that before treatment (P＜0.05). There were no significant differences in sperm DFI, sperm acrosome integrity rate and sperm spontaneous acrosome reaction rate between the two groups(P＞0.05). There were no significant differences in the fertilization rate, 2PN fertilization rate, cleavage rate, excellent embryo rate, implantation rate, clinical pregnancy rate, live birth rate, premature birth rate, abortion rate and neonatal malformation rate between the two groups (P＞0.05). A total of 103 fresh transplant cycles, 989 MII oocytes, 773 zygotes, 49 clinical pregnancies and 39 live births were obtained (including 17 in the TESA group and 22 in the PESA group). During a 3-month follow-up after birth, it was found that one newborn had cardiac abnormalities in the TESA group, while the other newborns had no abnormalities. Conclusion In OA patients, L-carnitine before TESA-ICSI and PESAICSI can improve the sperm quality, optimize clinical outcome, and the medication is safe. [ABSTRACT FROM AUTHOR]

Objective: To assess the role of testicular sperm in improving the success rates among patients who have repeated failed intracytoplasmic sperm injection cycles using ejaculated sperm. Design: A retrospective review of records. Materials and Methods: The study included 110 partners with repeated failed ICSI using ejaculate sperm were allocated into two groups retrospectively, one of them underwent testicular ICSI and the other underwent ejaculate ICSI. The two groups were well matched regarding their age of wife, age of husband, duration of marriage, history of previous marriage and previous children. Results: Showed that there was no statistically significant difference between patients with second ejaculate ICSI and patients with second testicular ICSI regarding the number of embryos and pregnancy rate (P- value>0.05). Patients underwent second testicular ICSI had a fertilization rate insignificantly higher than patients underwent second ejaculate ICSI. Conclusions: ICSI using testicular extracted sperm may be considered an alternative to using second ejaculated sperm in the treatment plan of infertile couples especially if increased SDF. We infer that increased SDF may be the main cause of repeated failures of ICSI using ejaculated sperm which remains to be proved by various studies. Higher pregnancy and live birth rates are associated more with higher testicular biopsy motility per drop, testicular biopsy concentration, Number of embryos and the fertilization rate. This study recommends that men with repeated failed ejaculate ICSI with high DNA fragmentation can gain benefit from testicular ICSI in the form of increased fertilization rate, number of embryos and live birth rate. [ABSTRACT FROM AUTHOR]

Purpose: The main objective of this opinion paper was to bring to light and enhance our understanding of the amount of double-strand DNA breaks in sperm and whether there is a threshold of no return when considering repair by the oocyte/embryo. Methods: A brief review of literature related to the theories proposed for the appearance of double-strand breaks in human spermatozoa. Further commentary regarding their detection, how oocytes or embryos may deal with them, and what are the consequences if they are not repaired. Finally, a strategy for dealing with patients who have higher levels of double-strand DNA breaks in sperm is proposed by reviewing and presenting data using testicular extracted sperm. Results: We propose a theory that a threshold may exist in the oocyte that allows either complete or partial DNA repair of impaired sperm. The closer that an embryo is exposed to the threshold, the more the effect on the ensuing embryo will fail to reach various milestones, including blastocyst stage, implantation, pregnancy loss, an adverse delivery outcome, or offspring health. We also present a summary of the role that testicular sperm extraction may play in improving outcomes for couples in which the male has a high double-strand DNA break level in his sperm. Conclusions: Double-strand DNA breaks in sperm provide a greater stress on repair mechanisms and challenge the threshold of repair in oocytes. It is therefore imperative that we improve our understanding and diagnostic ability of sperm DNA, and in particular, how double-strand DNA breaks originate and how an oocyte or embryo is able to deal with them. [ABSTRACT FROM AUTHOR]

Background: The combination of microsurgical testicular sperm extraction (mTESE) and intracytoplasmic sperm injection (ICSI) has become a common management option, with sperm motility being positively associated with successful outcomes. However, few studies have investigated whether the use of fresh or thawed gamete combinations affect clinical outcomes. Objectives: To determine whether the clinical outcomes of ICSI cycles using mTESE recovered testicular sperm of non-obstructive azoospermia (NOA) patients are affected by using fresh or thawed gametes. Material and Methods: A retrospective study was conducted of NOA patients who underwent mTESE between 2017 and 2020 at Monash IVF assisted reproductive clinics in Melbourne, Australia. The impact of gamete fresh/frozen status and sperm motility was investigated on clinical outcomes such as fertilization, blastocyst formation, clinical pregnancy, and live birth rates (LBRs). Results:A total of 103 NOA patients underwent mTESE, with a 65.1% successful surgical-sperm-retrieval. In total 56 patients contributed to 68 ICSI cycles, with a 35.1% fertilization rate and 25% LBR per embryo transfer. Compared with fresh testicular sperm, thawed testicular sperm did not affect clinical outcomes, including LBRs [16.7% vs 12.0%; odds ratio (OR) 0.68 (0.18–2.70)]. However, the use of thawed oocytes had a negative effect on fertilization rates [fresh-oocytes, 37.8%; vitrified-oocytes, 34.5%; OR 0.86 (0.02–0.48)]. Cycles using only motile sperm had a greater fertilization rate than those using a combination of motile and non-motile sperm (49.6% vs 37.2%, p ¡ 0.05). Importantly, when exclusively non-motile sperm were available (n = 26 cycles) their injection resulted in a very low fertilization rate (2.7%) and no live births were recorded. Conclusion: Micro-TESE is an effective treatment for NOA patients, with no clear advantage of using fresh over thawed sperm, however, the use of vitrified compared with fresh oocytes requires further investigation. Importantly, patients should be informed of the poor outcomes with the use of non-motile sperm in mTESE ICSI cycles.

IntroductionCryptozoospermia is defined by the World Health Organization (WHO) as the presence of isolated sperm cell in the ejaculate only identified after an extended microscopic search or after being pelleted. Although the number of spermatozoa is usually sufficient for intracytoplasmic sperm injection (ICSI), ICSI fails due to poor sperm quality in some cases. Contention remains regarding whether testicular sperm offers any advantage in this unique situation. At our tertiary referral center, we will offer patients a surgical sperm retrieval via conventional or microdissection testicular sperm extraction (microTESE) for men with cryptozoospermia and failed ICSI, or where ejaculated specimens are immotile or insufficient for ICSI. In this study, we sought to describe our experience and evaluate the predictors of success in cryptozoospermic patients who had microTESE at our center.MethodsWe retrospectively reviewed our electronic medical records for all patients with cryptozoospermia who underwent microTESE between 2007- 2021 for failed ICSI with ejaculated sperm or sperm quality deemed to be of insufficient quality for ICSI (e.g., nonmotile sperm). We evaluated demographics, preoperative lab results, pathology results, sperm retrieval rate (SRR) and ICSI outcomes.Results28 cryptozoospermic patients were identified. These patients underwent 37 unique microTESE. 22 of these men had failed previous ICSI treatment with ejaculated sperm, while the other 6 patients had ejaculated sperm with non-suitable quality for ICSI. None had genetic abnormalities. Successful retrieval of motile sperm suitable for ICSI was achieved in in 30 micro TESE procedures (SRR: 81.0%).14 out of 28 patients (50%) who underwent embryo transfer had positive pregnancy result, and 12/28 patients (42.8%) had successful live birth. The most common pathological pattern was hypospermatogenesis found in 65.3% (17/26). Fibrosis pathology was significantly higher in the negative pregnancy group. There were no postoperative complications noted.DisscussionThe use of testicular sperm in cryptozoospermic men with failed prior ICSI using ejaculated sperm has a high rate of pregnancy and live birth. While still controversial, our results suggest that surgical sperm retrieval is a viable option for these men with minimal risk of complications.

Herein, we introduced a novel individual sperm freezing device named SpermCD, which consists of a right angular cryopiece (RA‐Cryopiece, or "C") and a grooved petri dish ("D"). SpermCD allows embryologists to transfer sperm and perform ICSI on the same focal plane. Thirty‐five patients underwent single sperm cryopreservation using SpermCD, including four patients with non‐obstructive azoospermia (NOA), 14 patients with virtual azoospermia and 17 patients with cryptozoospermia. One hundred and twenty‐five cryopreserved spermatozoa from nine patients were thawed on the day of the oocyte retrieval and 121 spermatozoa were found, with a sperm recovery rate of 97.1 ± 4.6%. Sixty‐five MII oocytes from their spouse were injected with thawed sperm. Normal fertilization and high‐quality embryo rates were 68.0% ± 33.2% and 24.4% ± 22.2%. Nineteen transplantable embryos were formed after fertilization with frozen sperm, eight of which were transplanted in five couples, resulting in four successful deliveries. SpermCD is a simple and practical individual sperm freezing device. [ABSTRACT FROM AUTHOR]

Purpose: The use of testicular sperm is confined to patients with azoospermia, but there is evidence to support its use in males with poor semen parameters and/or previous intracytoplasmic sperm injection (ICSI) failures with ejaculated spermatozoa. We compared the aneuploidy rate and quality between embryos derived from ICSI cycles with ejaculated sperm (EJ-ICSI) and those from ICSI cycles using testicular spermatozoa (TT-ICSI) within the same couple. Methods: Retrospective study of 27 couples who first underwent an EJ-ICSI cycle that did not result in a livebirth and afterwards a TT-ICSI cycle. Only the two closer cycles of each couple were included. Preimplantation genetic test for aneuploidies (PGT-A) was performed in both ICSI cycles and classic parameters of embryo quality were assessed until blastocyst-stage. Results: A total of 375 embryos from 54 ICSI cycles were evaluated. Aneuploidy rate was measured by two different parameters. Patients undergoing TT-ICSI presented a similar aneuploidy rate as EJ-ICSI group: 30.7% (23.4–38.0) vs 26.8% (18.1–35.5) per inseminated oocytes (P>0.05), and 76.2% (66.2–86.2) vs 72.1% (59.1–85.2) per the total number of biopsied embryos (P>0.05), respectively. Further, the good-quality blastocyst rate per correctly fertilized oocyte was significantly higher in TT-ICSI group (33.6% (30.4–36.9)) than EJ-ICSI group (24.2% (20.3–28.0)) (P<0.001). Conclusions: Switching to testicular sperm for ICSI yielded better-quality blastocysts without affecting the chromosomal load of the embryos in non-azoospermic couples with a previous unsuccessful ICSI using ejaculated sperm. This strategy is a good option for couples seeking a livebirth who do not want to use donor sperm. [ABSTRACT FROM AUTHOR]

To assess the feasibility of repeated sperm recovery in patients with non-obstructive azoospermia (NOA), as little is known about the extraction rate in repeated microdissection testicular sperm extraction (microTESE) in these patients. A total of 134 men with NOA had their first sperm recovery between January 2013 and February 2020. Repeated microTESE had been done mostly for patients with a successful initial retrieval. In the 323 procedures performed on the 134 men with NOA, sperm could be retrieved in 236 procedures (73.1%). A total of 88, 61 and 40 men underwent two, three and four sperm retrievals, respectively. In these cycles, sperm could be extracted in 65 (73.9%), 53 (86.9%) and 37 (92.5%) men, respectively. During the first microTESE procedure, sperm could be extracted in 81 (60.4%) men with NOA. In all, the success rate was significantly different between subgroups, showing highest rate in hypospermatogenesis cases (95.6%), followed by maturation arrest (58.5%), and Sertoli cell-only syndrome (56.0%). However, this difference was not significant at the third and fourth repeated microTESE. The FSH levels and testicular volume were among the noticeable factors affecting success of sperm retrieval. The duration between the first and second biopsies significantly increased the success rate by a factor of 1.3-fold/month; however, afterwards, the duration did not play any role in the success of microTESE. The success of previous trial significantly increased the probability of success by 10.1-fold in the second trial, 5.6-fold in the third trial, and 16.5 folds in the fourth. Repeated MD -TESE ensures a high sperm recovery rate in patients with NOA. These data also show that when no spermatozoa can be obtained after thawing cryopreserved testicular sperm for ICSI in NOA patients, a repeat microTESE procedure can be planned. ICSI: intracytoplasmic sperm injection; IVF: in vitro fertilisation; MA: maturation arrest; (N)OA: (non-)obstructive azoospermia; OR: odds ratio; SCOS, Sertoli cell-only syndrome; SRR: spermatozoa retrieval rate; (micro)TESE: (microdissection) testicular sperm extraction [ABSTRACT FROM AUTHOR]

Background: Most of data available in the literature reported the sperm retrieval rate and limited intracytoplasmic sperm injection (ICSI) results of microdissection testicular sperm extraction (micro-TESE) in non-obstructive azoospermia (NOA) patients with different etiologies. Unfortunately, there is currently a lack of comprehensive data to guide clinicians in conducting comprehensive consultations with NOA patients. Objectives: To obtain more comprehensive evidence-based data and clinical outcomes for better consultation of NOA patients who opted to undergo micro-TESE combined with ICSI-IVF Methods: It was a retrospective study involved 968 NOA patients underwent micro-TESE during January 2015 to December 2019. Embryological, clinical, and live birth outcomes were demonstrated comprehensively and three kinds of stratification analyses were performed based on ICSI-IVF cycles using frozen and fresh sperm, different etiologies of NOA and various amounts of sperm retrieved. Results: The sperm retrieval rate was 44.6%, and ICSI was performed in 299 couples leading to 150 clinical pregnancies and 140 live-birth deliveries. The clinical pregnancy rate (CPR) was 50.17%, and the cumulative live birth rate (LBR) was 46.82%, and the low birth defects rate was 1.43%. No significant difference was observed about cumulative LBR in frozen sperm group and fresh sperm group (47.5% vs 42.9%, P> 0.05). NOA patients with AZFc microdeletions had the lowest rate of a high-score embryo on day 3 (4.4%, P <0.05) and the lowest cumulative LBR (19.4%, P <0.05). NOA patients with lower sperm count (having fewer than 20 sperms retrieved) had significantly lower cumulative LBR than those with higher sperm count (having more than 20 sperms retrieved) (28.1% vs 51.9%, P <0.05). Conclusions: For those NOA patients who stepped in ICSI-IVF cycles, the cumulative LBR was 46.82%. No significant difference was indicated in the LBR between ICSI-IVF cycles using frozen or fresh testicular sperm. Compared to other etiologies, NOA caused by AZFc microdeletions have the poorest embryological and clinical outcomes. Patients with less testicular sperm retrieved have poorer embryological and clinical outcomes. [ABSTRACT FROM AUTHOR]

BackgroundMost of data available in the literature reported the sperm retrieval rate and limited intracytoplasmic sperm injection (ICSI) results of microdissection testicular sperm extraction (micro-TESE) in non-obstructive azoospermia (NOA) patients with different etiologies. Unfortunately, there is currently a lack of comprehensive data to guide clinicians in conducting comprehensive consultations with NOA patients.ObjectivesTo obtain more comprehensive evidence-based data and clinical outcomes for better consultation of NOA patients who opted to undergo micro-TESE combined with ICSI-IVFMethodsIt was a retrospective study involved 968 NOA patients underwent micro-TESE during January 2015 to December 2019. Embryological, clinical, and live birth outcomes were demonstrated comprehensively and three kinds of stratification analyses were performed based on ICSI-IVF cycles using frozen and fresh sperm, different etiologies of NOA and various amounts of sperm retrieved.ResultsThe sperm retrieval rate was 44.6%, and ICSI was performed in 299 couples leading to 150 clinical pregnancies and 140 live-birth deliveries. The clinical pregnancy rate (CPR) was 50.17%, and the cumulative live birth rate (LBR) was 46.82%, and the low birth defects rate was 1.43%. No significant difference was observed about cumulative LBR in frozen sperm group and fresh sperm group (47.5% vs 42.9%, P>0.05). NOA patients with AZFc microdeletions had the lowest rate of a high-score embryo on day 3 (4.4%, P

Studies have explored the assisted reproductive technology (ART) outcomes of Y-chromosome azoospermia factor c (AZFc) microdeletions, but the effect of sperm source on intracytoplasmic sperm injection (ICSI) remains unknown. To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions, we searched Embase, Web of Science, and PubMed to conduct a systematic review and meta-analysis. The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group (risk ratio: 0.97, 95% confidence interval [CI]: 0.73–1.28, P = 0.82). The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group (risk ratio: 1.06, 95% CI: 0.54–2.06, P = 0.87). The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group (risk ratio: 1.24, 95% CI: 0.66–2.34, P = 0.50). Inevitable heterogeneity weakened our results. However, our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes, representing a meaningful result for clinical treatment. More properly designed studies are needed to further confirm our conclusions.

The clinical applications of donated gametes are approved in many countries; however, attitudes toward its application and national legislation in some countries are challenging. The purpose of this study is to report a healthy live birth produced by vitrified‐warmed oocytes and frozen‐thawed testicular sperms to avoid sperm donation. [ABSTRACT FROM AUTHOR]

Human autologous sperm freezing involves ejaculated sperm, and testicular or epididymal puncture sperm freezing, and autologous sperm freezing is widely used in assisted reproductive technology. In previous studies, researchers have tried to cryopreserve sperm from mammals (rats, dogs, etc.) using a −80°C freezer and have achieved success. It is common to use liquid nitrogen vapor rapid freezing to cryopreserve human autologous sperm. However, the operation of this cooling method is complicated, and the temperature drop is unstable. In this study, we compared the quality of human ejaculation and testicular sperm after liquid nitrogen vapor rapid freezing and −80°C freezing for the first time. By analyzing sperm quality parameters of 93 ejaculated sperm and 10 testicular sperm after liquid nitrogen vapor rapid freezing and −80°C freezing, we found reactive oxygen species (ROS) of sperm of the −80°C freezer was significantly lower than liquid nitrogen vapor rapid freezing. Regression analysis showed that progressive motility, ROS, and DNA fragmentation index (DFI) in post-thaw spermatozoa were correlated with sperm progressive motility, ROS, and DFI before freezing. For the freezing method, the −80°C freezer was positively correlated with the sperm progressive motility. Among the factors of freezing time, long-term freezing was negatively correlated with sperm progressive motility and ROS. Although freezing directly at −80°C freezer had a slower temperature drop than liquid nitrogen vapor rapid freezing over the same period, the curves of the temperature drop were similar, and slight differences in the freezing point were observed. Furthermore, there were no statistically significant differences between the two methods for freezing testicular sperm. The method of direct −80°C freezing could be considered a simplified alternative to vapor freezing for short-term human sperm storage. It could be used for cryopreservation of autologous sperm (especially testicular sperm) by in vitro fertilization centers.Clinical Trial Registration: (website), identifier (ChiCTR2100050190).

Summary: This study examined the effect of a cryoprotectant with and without pentoxifylline supplementation on the motility and viability of human testicular sperm, both before and after freezing. Testicular samples were obtained from 68 patients with azoospermia who came to the Andrology Service of West China Second University Hospital, Sichuan University, for testicular biopsies from December 2019 to April 2020. All patients were assigned randomly to two groups: experimental, whose testicular sperm were added to the cryoprotectant with pentoxifylline, and the control, whose testicular sperm were added to the cryoprotectant without pentoxifylline. Both groups used the same freezing and thawing methods. Testicular sperm motility in the experimental group was significantly higher than that of the control group, both before and after cryopreservation. The recovery rate of sperm motility in the experimental group was significantly higher than that of the control group. The percentage of samples with motile testicular sperm in the experimental group was significantly higher than that of the control group after thawing. Sperm viability was unchanged between the experimental and control groups, both before and after freezing. Overall, a pentoxifylline-supplemented cryoprotectant can significantly improve the motility of testicular sperm before and after cryopreservation. [ABSTRACT FROM AUTHOR]

Human autologous sperm freezing involves ejaculated sperm, and testicular or epididymal puncture sperm freezing, and autologous sperm freezing is widely used in assisted reproductive technology. In previous studies, researchers have tried to cryopreserve sperm from mammals (rats, dogs, etc.) using a −80°C freezer and have achieved success. It is common to use liquid nitrogen vapor rapid freezing to cryopreserve human autologous sperm. However, the operation of this cooling method is complicated, and the temperature drop is unstable. In this study, we compared the quality of human ejaculation and testicular sperm after liquid nitrogen vapor rapid freezing and −80°C freezing for the first time. By analyzing sperm quality parameters of 93 ejaculated sperm and 10 testicular sperm after liquid nitrogen vapor rapid freezing and −80°C freezing, we found reactive oxygen species (ROS) of sperm of the −80°C freezer was significantly lower than liquid nitrogen vapor rapid freezing. Regression analysis showed that progressive motility, ROS, and DNA fragmentation index (DFI) in post-thaw spermatozoa were correlated with sperm progressive motility, ROS, and DFI before freezing. For the freezing method, the −80°C freezer was positively correlated with the sperm progressive motility. Among the factors of freezing time, long-term freezing was negatively correlated with sperm progressive motility and ROS. Although freezing directly at −80°C freezer had a slower temperature drop than liquid nitrogen vapor rapid freezing over the same period, the curves of the temperature drop were similar, and slight differences in the freezing point were observed. Furthermore, there were no statistically significant differences between the two methods for freezing testicular sperm. The method of direct −80°C freezing could be considered a simplified alternative to vapor freezing for short-term human sperm storage. It could be used for cryopreservation of autologous sperm (especially testicular sperm) by in vitro fertilization centers. Clinical Trial Registration: (website), identifier (ChiCTR2100050190). [ABSTRACT FROM AUTHOR]

The aim of the study is to compare the reproductive outcomes of different sperm selection techniques: density gradient centrifugation (DGC), testicular sperm (Testi), physiological ICSI (PICSI), and magnetic-activated cell sorting (MACS) in abnormal sperm DNA fragmentation (SDF) ICSI patients. A randomized controlled trial included 302 patients with abnormal SDF undergoing ICSI where they were randomized into 4 groups: a control group of DGC (n= 72), Testi (n=73), PICSI (n=78), and MACS (n=79). Results showed no significant differences in the male age, female age, or SDF between the four groups. Testi group had significantly lower cleavage and blastulation rates compared to PICSI, DGC, or MACS groups (p =0.001). For the high-quality blastocysts, DGC and MACS groups had significantly higher rate than the Testi group (p =0.014). The highest pregnancy rate was scored for the PICSI group (69.6%), while the lowest pregnancy rate was scored for the DGC group (51.4%) with (p =0.025). The PICSI group showed a significantly higher implantation rate compared to the other groups (p =0.003). Regarding the ongoing pregnancy rate, the significant difference was observed between the PICSI (62.8%) and MACS (62%) vs. DGC (45.8%). Besides, no significant differences were found in the miscarriage rates between the four groups. In conclusion, PICSI and MACS along with DGC showed significant improvement in embryological and clinical outcome over testicular sperm or sperm processed by DGC alone in patients with abnormal SDF Registration number: NCT04482517 [ABSTRACT FROM AUTHOR]

: To review the debate about the routine use of cryopreserved testicular sperm for intracytoplasmic sperm injection (ICSI) from patients with non-obstructive azoospermia (NOA), as some authors suggest repeating sperm retrieval in such cases due to poorer ICSI results when frozen–thawed testicular sperm is used compared with fresh sperm. : A systematic literature review was performed in August 2020 using the Medical Literature Analysis and Retrieval System Online (MEDLINE), Web of Science databases and the Excerpta Medica dataBASE (EMBASE), and we included 26 studies that were considered eligible for this systematic review. : In all, 1189 publications were screened and 26 articles were included in the systematic review. Three meta-analysis reviews were included and they all concluded that the use of fresh and frozen sperms for ICSI from patients with NOA showed comparable fertilisation and pregnancy rates. : The use of frozen testicular sperm from men with NOA results in fertilisation and clinical pregnancy rates similar to those of fresh sperm. This may encourage fertility centres to use frozen testicular sperm samples, as this policy has certain advantages that would help with organising their workflow. Abbreviations: CPR: clinical pregnancy rate; 2PN%: two pronuclei % fertilisation rate; ICSI: intracytoplasmic sperm injection; NOA: non-obstructive azoospermia; OA, obstructive azoospermia; SCO: Sertoli cell-only syndrome; (micro-)TESE: (microsurgical) testicular sperm extraction [ABSTRACT FROM AUTHOR]

Sperm DNA fragmentation (SDF) has been linked with male infertility, and previous studies suggest that SDF can have negative influence on pregnancy outcomes with assisted reproduction. We performed a retrospective review of consecutive couples with a high SDF level that had intracytoplasmic sperm injection (ICSI) using testicular sperm (T-ICSI). We compared the T-ICSI outcomes to that of two control groups: 87 couples with failed first ICSI cycle and who had a second ICSI cycle using ejaculated sperm (Ej-ICSI), and 48 consecutive couples with high sperm chromatin structure assay (SCSA)-defined SDF (>15%) that underwent an ICSI cycle using ejaculated sperm after one or more failed ICSI cycles (Ej-ICSI-high SDF). The mean number of oocytes that were retrieved and the total number of embryos were not different among the three groups. The mean number of transferred embryos in the T-ICSI group was higher than the Ej-ICSI group but not significantly different than the Ej-ICSI-high SDF group (1.4, 1.2, and 1.3, respectively, P < 0.05). Clinical pregnancy rate in the T-ICSI group was not significantly different than the Ej-ICSI and Ej-ICSI-high SDF groups (48.6%, 48.2%, and 38.7%, respectively, P > 0.05). No significant difference was found in live birth rate when comparing T-ICSI to Ej-ICSI and Ej-ICSI-high SDF groups. The results suggest that pregnancy outcomes and live birth rates with T-ICSI are not significantly superior to Ej-ICSI in patients with an elevated SCSA-defined sperm DNA fragmentation and prior ICSI failure(s).

What are the cumulative clinical pregnancy rates (CPR) and live births rates (LBR) in intracytoplasmic sperm injection (ICSI) cycles using testicular motile compared with immotile spermatozoa, obtained from testicular sperm aspiration (TESA) or extraction (TESE)? A retrospective analysis of ICSI cycles using TESA or TESE over a period of 7 years. Cycles were divided into two groups according to the motility of the retrieved spermatozoa: Group A consisted of couples with motile spermatozoa; Group B of couples with immotile spermatozoa. Group B was subdivided into two groups: B1 consisted of couples with motile spermatozoa and B2 with immotile spermatozoa after the addition of pentoxifylline. No differences in CPR and LBR per transfer was found between the study groups after fresh embryo transfer. No pregnancies were achieved by vitrified–warmed embryo transfer in group B2. Fertilization rates decreased when using immotile spermatozoa (64.4%, 56%, 37.9%, for groups A, B1 and B2, respectively, P < 0.001). Top-quality embryo rates were higher in groups A and B1 compared with B2 (40.7% and 40.1% versus 19.1%, respectively, P = 0.015). Cumulative CPR (53%, 41.7%, 13.6% for groups A, B1 and B2, respectively, P = 0.005) and LBR (42.4%, 30%, 13.6% for groups A, B1 and B2, respectively P = 0.03) per oocyte retrieval was significantly higher when using motile spermatozoa compared with motile or immotile spermatozoa after adding pentoxifylline. Although fertilization, top-quality embryo rates, cumulative CPR and LBR decreased when using immotile spermatozoa, ICSI is still valid; therefore, it should be considered and offered to couples before embarking on a donor sperm insemination cycle, or cryopreserving oocytes for future additional testicular sperm retrieval. [ABSTRACT FROM AUTHOR]

The most common reason for in vitro fertilization (IVF) cycle cancelation is a lack of quality gametes available for intracytoplasmic sperm injection (ICSI). Here we present the successful fertility treatment of the couple affected by obstructive azoospermia combined with suboptimal response to controlled ovarian stimulation. Since the conventional approach appeared ineffective to overcome both partnersˈ specific problems, the targeted interventions, namely, (1) pharmacological enhancement of sperm motility and (2) polarized light microscopy (PLM)-guided optimization of ICSI time, were applied to rescue the cycle with only immature oocytes and immotile testicular sperm retrieved. The treatment with theophylline aided the selection of viable spermatozoa derived from cryopreserved testicular tissue. When the traditional stimulation protocol failed to produce mature eggs, non-invasive spindle imaging was employed to adjust the sperm injection time to the maturational stage of oocytes extruding a polar body in vitro. The fertilization of 12 late-maturing oocytes yielded 5 zygotes, which all developed into blastocysts. One embryo was transferred into the uterus on day 5 post-fertilization, and another 3 good quality blastocysts were vitrified for later use. The pregnancy resulted in a full-term delivery of a healthy child. This case demonstrates that the individualization beyond the standard IVF protocols should be considered to maximize the chance of poor-prognosis patients to achieve pregnancy with their own gametes. [ABSTRACT FROM AUTHOR]

Background: In patients with azoospermia, pregnancy can be achieved after surgical techniques using sperm retrieved from the testis or epididymis, which can impact on DNA integrity and epigenetics. DNA of the fetus and placenta is equally derived from both parents; however, genes important for placental development are expressed from the paternal alleles. Therefore, the origin of sperm may affect fetal and placental development. Objectives: To investigate whether first‐trimester trajectories of embryonic and placental development of pregnancies conceived after intracytoplasmic sperm injection (ICSI) with testicular sperm extraction (TESE) or microsurgical epididymal sperm aspiration (MESA), are different from pregnancies after ICSI with ejaculated sperm or natural conceptions. Materials and methods: A total of 147 singleton ICSI pregnancies, including pregnancies conceived after TESE (n = 23), MESA (n = 25) and ejaculated sperm (n = 99), and 380 naturally conceived and 140 after IVF treatment without ICSI were selected from the prospective Rotterdam periconception cohort. Crown‐rump length (CRL), embryonic volume (EV), Carnegie stages, and placental volume (PV) at 7, 9, and 11 weeks of gestation were measured using 3D ultrasound and virtual reality technology. Results: Linear mixed model analysis showed no differences in trajectories of CRL, EV, and Carnegie stages between pregnancies conceived after ICSI with testicular, epididymal, and ejaculated sperm. A significantly positive association was demonstrated for PV between pregnancies conceived after TESE‐ICSI (adjusted beta: 0.28(95%CI: 0.05‐0.50)) versus ICSI with ejaculated sperm. Retransformation to original values showed that the PV of pregnancies after TESE‐ICSI is 14.6% (95%CI: 1.4%‐25.5%) larger at 11 weeks of gestation compared to ICSI pregnancies conceived with ejaculated sperm. Discussion and Conclusion: Here we demonstrate that the first‐trimester growth trajectory of the placenta is increased in pregnancies conceived after TESE‐ICSI compared to those conceived after ICSI with ejaculated sperm. Findings are discussed in the light of known differences in sperm DNA integrity, epigenetics, and placental gene expression. [ABSTRACT FROM AUTHOR]

Sperm DNA fragmentation (SDF) has been linked with male infertility, and previous studies suggest that SDF can have negative influence on pregnancy outcomes with assisted reproduction. We performed a retrospective review of consecutive couples with a high SDF level that had intracytoplasmic sperm injection (ICSI) using testicular sperm (T-ICSI). We compared the T-ICSI outcomes to that of two control groups: 87 couples with failed first ICSI cycle and who had a second ICSI cycle using ejaculated sperm (Ej-ICSI), and 48 consecutive couples with high sperm chromatin structure assay (SCSA)-defined SDF (>15%) that underwent an ICSI cycle using ejaculated sperm after one or more failed ICSI cycles (Ej-ICSI-high SDF). The mean number of oocytes that were retrieved and the total number of embryos were not different among the three groups. The mean number of transferred embryos in the T-ICSI group was higher than the Ej-ICSI group but not significantly different than the Ej-ICSI-high SDF group (1.4, 1.2, and 1.3, respectively, P < 0.05). Clinical pregnancy rate in the T-ICSI group was not significantly different than the Ej-ICSI and Ej-ICSI-high SDF groups (48.6%, 48.2%, and 38.7%, respectively, P > 0.05). No significant difference was found in live birth rate when comparing T-ICSI to Ej-ICSI and Ej-ICSI-high SDF groups. The results suggest that pregnancy outcomes and live birth rates with T-ICSI are not significantly superior to Ej-ICSI in patients with an elevated SCSA-defined sperm DNA fragmentation and prior ICSI failure(s). [ABSTRACT FROM AUTHOR]

Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation (SDF) in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology (ART), an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection (Testi-ICSI). In this article, we used a SWOT (strengths, weaknesses, opportunities, and threats) analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval.

Abstract Research question Does the adapted carrier Cryoplus improve the quality of cryopreserved spermatozoa compared with the use of conventional containers, and what is the effect of the adapted carrier on clinical outcomes? Design Semen samples from 27 cases of oligozoospermia were used to investigate whether the adapted carrier improved cryopreserved sperm quality compared with the use of 0.25-ml straws and 2-ml cryogenic vials. Thirty testicular sperm samples were used to study the quality of testicular spermatozoa cryopreserved in the adapted carrier. The retrospective study included a further 104 men with azoospermia to investigate the clinical outcomes of testicular spermatozoa cryopreserved with the adapted carriers. Men with mostly obstructive azoospermia were included in this study. Results The adapted carrier improved cryopreserved spermatozoa motility of semen samples compared with 2-ml cryogenic vials but not compared with 0.25-ml straws. No differences were found in cryopreserved sperm DNA fragmentation among the three carriers. Fertilization and good-quality embryo rates were similar in ICSI cycles using fresh or cryopreserved testicular spermatozoa. Additionally, no difference was evident between frozen–thawed embryo transfer cycles using fresh or cryopreserved testicular spermatozoa in clinical pregnancy, implantation, miscarriage, live birth rates or birth weight. Conclusions The adapted carrier improved the cryopreserved sperm motility compared with the effects of 2-ml cryogenic vials. The outcomes of intracytoplasmic sperm injection and frozen-thawed embryo transfer outcomes indicate that testicular spermatozoa cryopreserved using the adapted carrier is not inferior to fresh testicular spermatozoa. The use of the adapted carrier for cryopreserving human testicular spermatozoa especially from obstructive azoospermia is simple and effective. [ABSTRACT FROM AUTHOR]

Background: Miscarriage and take‐home baby are the most important issues to patients with cryptozoospermia in receiving intracytoplasmic sperm injection (ICSI). The ICSI usually use ejaculated or testicular sperm. Unfortunately, no synthesized evidence reported miscarriage and take‐home baby rate between the two sperm sources. Objectives: This study aimed to compare the miscarriage and take‐home baby rate of ICSI using testicular and ejaculated sperm in patient with cryptozoospermia. Materials and methods: We conducted meta‐analyses that were based on data from Cochrane library, Ovid, PubMed, ScienceDirect, Scopus, and Web of Science. The pooled analyses used risk ratio (RR) in random‐effects model. Sensitivity analyses by subgrouping were completed to explore the associations between mean age and outcome. Results: This study identified 331 potential citations and included four cohort studies for qualitative and quantitative synthesis. The four studies involved 331 patients with 479 ICSI cycles. The results showed no significant difference in miscarriage between testicular sperm group and ejaculated sperm group (RR = 1.06, 95% CI 0.48–2.35, p = 0.88). Yet, take‐home babies per embryo transfer in testicular sperm group (53/226, 23.45%) was more than ejaculated sperm group (59/429, 13.75%) (RR = 1.72, 95% CI 1.21–2.44, p = 0.002). Similar results can be found in take‐home babies per ICSI cycle (RR = 1.77, 95% CI 1.28–2.44, p = 0.0005), especially in younger couple (RR = 1.93, 95% CI 1.11–3.34, p = 0.02). No small study bias was detected in the analyses. Discussion: This study found that testicular sperm has more advantage for ICSI in patients with cryptozoospermia, especially in younger couple. These findings may help guide us when deciding the optimal method of sperm harvest for men with cryptozoospermia. Conclusion: Comparing to ejaculated sperm, testicular sperm showed benefits for take‐home baby rate, but not for miscarriage in patients with cryptozoospermia. [ABSTRACT FROM AUTHOR]

Purpose: The aims of this study were (1) to evaluate clinical outcomes after ICSI cycles using surgically recovered sperm and (2) to assess the influence of maternal age on those outcomes.Methods: A retrospective cohort study of 24,763 IVF cycles of fresh autologous oocytes and ICSI using surgically recovered sperm reported to the SART CORS database from 2004 to 2015.Results and conclusions: Older women had significantly longer stimulation (p < 0.001), a lower number of oocytes retrieved (p < 0.001), a lower number of 2PN zygotes (p < 0.001), a lower chance of having a blastocyst transferred (p < 0.001), and a higher number of fresh embryos transferred (p < 0.001). There was no significant association between the number of 2PNs per oocyte retrieved and maternal age (p = 0.214). Both clinical pregnancy rates and live birth rates (LBR) decreased with advanced maternal age (p < 0.001). LBR ranged from 50.4% in women < 30 to 7.2% in women > 42 years, and for cleavage-stage transfers, the LBR ranged from 47.3% in women< 30 to 6.3% in women > 42 years. There were no differences in gestational age at delivery, proportion of term deliveries, preterm deliveries, neonatal birth weight < 2500 g, neonatal birth weight > 4000 g and average birthweight of neonates for singleton pregnancies according to age. For twin pregnancies, women < 30 years had significantly higher number of live births, term deliveries, and lower preterm deliveries than older women. There was a similar number of female (6051) and male neonates (5858; p = 0.2). Overall, pregnancy outcomes with ICSI using surgically recovered sperm are reassuring and comparable to those of ICSI with ejaculated sperm. [ABSTRACT FROM AUTHOR]

Purpose: Sperm play an essential role in embryonic genome activation and embryonic progression to blastocyst. In the present work, we focus on development of embryos created as a result of ICSI with testicular or epididymal sperm from azoospermic males and compare this to outcomes from normospermic males. The objective of this study was to determine if sperm origin influences clinical outcomes, the kinetics of embryo development, or the incidence of cleavage anomalies and multinucleation.Methods: A total of 93 consecutive intracytoplasmic sperm injection cycles (ICSI) performed for 83 couples were included in this study. Observations were made on 594 fertilized oocytes cultured in the EmbryoScope using time-lapse microscopy (TLM). Epididymal sperm (n = 29) cycles or surgically retrieved sperm from the testis (TESE; n = 37 cycles) of men with either obstructive (OA) or non-obstructive azoospermia (NOA) were used to inject oocytes. A further 27 ICSI cycles were performed using ejaculated sperm from normospermic males, designated as our control sperm (CS) group. Kinetic data and cycle outcomes were retrospectively analyzed.Results: The clinical pregnancy rate was not different between the three groups (TESE 51.4%, PESA 57.7%, and CS 59.3%). A non-significant decrease was observed in both implantation (30.9%) and live birth rate (43%) with TESE as compared to PESA (35.3%, 58%, respectively) and CS groups (45.1%, 56%, respectively). Failure to compact was significantly higher amongst TESE-NOA embryos (35.2%; P < 0.001) as compared to TESE-OA (4%), PESA (9%), and CS (3.8%) embryos. The two points at which TESE-derived embryos (both NOA and OA) behaved most differently from PESA and CS embryos was at cc2 (t3-t2; time to initiation of the second cell cycle) and tSB (time to start of blastulation). A significantly lower percentage of TESE embryos exhibited kinetics typically ascribed to high quality embryos with the greatest developmental potential. Finally, the incidence of direct uneven cleavage (DUC) was observed to be significantly higher after ICSI with sperm retrieved from azoospermic males.Conclusions: TLM allowed a more in depth comparison of paternal influence on embryo morphokinetics and helped to identify specific differences in cell cycle kinetics. TESE-NOA embryos exhibited a higher incidence of compaction failure. [ABSTRACT FROM AUTHOR]

Purpose: Recent studies have shown that improved clinical outcomes can be achieved by transferring blastocysts rather than cleavage-stage embryos. However, blastocyst transfer is not performed in all patients. The aim of this study was to compare clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles using testicular sperm (TE) with those of ICSI cycles using ejaculated sperm (EJ).Methods: ICSI was performed using EJ in 141 cycles and TE in 37 cycles. Embryos were cultured for 5 days. The quality of embryos was assessed on days 3 and 5 before embryo transfer.Results: Fertilization rate was 77.3% in the EJ group and 69.6% in the TE group (p < 0.05). The good-quality embryos on day 3 and 5 were not different between the EJ and TE groups. Embryos did not develop to blastocyst stage in 7 cycles of the EJ group (5.0%) and 2 cycles of the TE group (5.4%). There were no significant differences in blastocyst formation and blastocyst quality (46.1% vs. 47.5% and 5.7% vs 5.8%, respectively) on day 5 between both groups. Embryos were transferred in all cycles. Implantation (22.8 vs. 24.7%), clinical pregnancy (44.7 vs. 43.2%), miscarriage (21.7 vs. 33.3%), and delivery (76.5 vs. 66.7%) did not differ between EJ group and TE group. Clinical outcomes of ICSI were not different between the EJ and TE groups.Conclusions: In conclusion, the potential of testicular sperm supporting embryonic development to blastocysts is comparable to that of ejaculated sperm. Therefore, this study suggests that blastocyst transfer can be a very useful assisted reproductive technique in the ICSI cycles that require the use of testicular sperm, and the clinical outcomes of the cycles are comparable to those of ICSI cycles using ejaculated sperm. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the safety and efficacy of percutaneous epididymal and testicular sperm aspiration as a diagnostic technique to confirm sperm production and as a therapeutic technique to harvest sperms for use in the intracytoplasmic sperm injection and the indications for performing testicular biopsy in azoospermic infertile males. Thirty married patients were included in this prospective study from February 2011 to December 2012 seen in Basrah General Hospital. Their age ranged from 20 to 40 years. All patients underwent full medical examination with laboratory tests which included seminal fluid analysis, serum leutinising hormone (LH), follicular stimulating hormone (FSH), testosterone, and prolactin in addition to color Doppler ultrasonography of the scrotum. Patients with history of undescended testes, varicocele, & testicular pathology were excluded from this study. All patients showed normal physical examination with normal secondary sexual characters. The external genitalia were normal with normal sizes of their testes. The percutaneous epididymal and testicular sperm aspirations were positive in 12 out of 30 patients (40%). The rest had negative aspirations (60%). The testicular biopsy which performed in the patients with negative aspiration showed normal germinal epithelium with mature spermatozoa in only 5 patients out of 18 (28%) while the rest 13 patients had spermatogenic arrest (72%). In conclusion, percutaneous epididymal and testicular sperm aspiration has been found helpful as a diagnostic technique for patients with non-reconstructable azoospermia. It is a minimally invasive sperm retrieval technique and appears to be an effective alternative to microsurgical epididymal sperm aspiration, which is more invasive and costly. It is less invasive than testicular biopsy and preferably performed as a first step procedure in an attempt to obtain sperms for both diagnostic and therapeutic purposes.

Purpose: The purpose of this study is to analyze the sperm morphology of a Chinese man affected with multiple morphological abnormalities of the sperm flagella (MMAF) and observe the intracytoplasmic sperm injection (ICSI) outcome.Methods: A Chinese man was diagnosed with multiple morphological abnormalities of the sperm flagella by semen analysis and electron microscopy. Testicular spermatozoa were injected intracytoplasmically, and the following ICSI results were observed.Results: All the spermatozoa from his ejaculate were immotile and morphologically abnormal in the flagellum. In transmission electron microscopy assays, most spermatozoa showed disorganized fibrous sheath, accompanied by distortion of various cytoskeletal components, and missing of the central pair microtubules. Testicular sperm was injected to the oocytes in two ICSI cycles, with fertilization rates of 45.5 and 40.0%. Finally, a healthy female infant was delivered at the second ICSI cycle.Conclusions: Fertilization and pregnancy could be achieved by intracytoplasmic sperm injection, regardless of severe flagellar defects. ICSI is effective for MMAF-affected man, and testicular sperm is an alternative when no motile sperm is available. [ABSTRACT FROM AUTHOR]

Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation (SDF) in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology (ART), an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection (Testi-ICSI). In this article, we used a SWOT (strengths, weaknesses, opportunities, and threats) analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval. [ABSTRACT FROM AUTHOR]

In 1992 and subsequently, several reports indicated that ICSI was a successful technique to achieve clinical pregnancy and live birth using spermatozoa with severely impaired characteristics. The initial optimism over the ability of ICSI to overcome significant sperm abnormalities was later tempered by the findings of more recent publications suggesting that some sperm deficits may not be as effectively treated with ICSI. In search for effective treatment for couples with severe male factor, a number of small retrospective and prospective studies have reported high pregnancy and live birth rates using testicular sperm for men with necrozoospermia, cryptozoospermia and oligozoospermia with or without elevated sperm DNA damage. Although the data suggest that there may be some benefit in performing testicular sperm retrieval (TSR)-ICSI in select groups of non-azoospermic infertile men, there are potential risks involved with TSR. Clinicians should balance these risks prior to the recommendation of TSR-ICSI on the result of a semen analysis or sperm DNA test alone. Careful evaluation and management of male factor infertility is important. The use of TSR-ICSI in the absence of specific sperm DNA defects is still experimental. [ABSTRACT FROM AUTHOR]

Sperm DNA fragmentation ( SDF) is used in assisted reproductive technology ( ART) programs as an indicator for sperm quality, although there is still a lack of consensus as to its clinical utility. In this retrospective study, we examined intracytoplasmic sperm injection ( ICSI) outcomes of 1924 infertile patients who underwent SDF analysis using the sperm chromatin integrity test. ART patients were classified as having low [DNA fragmentation index ( DFI) <29%] or high SDF ( DFI ≥29%) and by whether or not an intervention [physiological intracytoplasmic sperm injection ( PICSI), intracytoplasmic morphologically selected sperm injection ( IMSI), testicular sperm extraction ( TESE)/testicular sperm aspiration ( TESA), frequent ejaculation] was performed. High SDF patients who did not have an intervention had a lower fertilization rate and poorer clinical outcomes from blastocyst transfers as compared with low SDF patients; the fertilization rate was 66.0% vs. 70.2% ( p = 0.042), single embryo transfer ( SET) fetal heart pregnancy rate was 28.5% vs. 45.2% ( p = 0.042), and SET live birth rate was 24.9% vs. 40.6% ( p = 0.060), respectively. Furthermore, high SDF patients who had an intervention had significantly improved blastocyst transfer outcomes, similar to those of low SDF patients; the SET live birth rate for high SDF intervention patients was 43.8% as compared with 24.9% for high SDF no intervention patients ( p = 0.037) and 40.6% for low SDF patients ( p = 0.446). Analysis of the three main intervention subgroups for high SDF patients revealed that TESE/ TESA patients had the highest SET live birth rate; in comparison with 24.2% for high SDF patients who did not have an intervention, PICSI patients had 38.3% ( p = 0.151), IMSI patients had 28.7% ( p = 0.680), and TESE/ TESA patients had 49.8% ( p = 0.020). Our data suggest that SDF results indicate ICSI outcomes and that patients who have high SDF benefit from an intervention. [ABSTRACT FROM AUTHOR]

Conventional sperm freezing methods perform best when freezing sperm samples containing at least hundreds of spermatozoa. In this severe male factor infertility case series, we examined the reproductive outcomes in 12 intracytoplasmic sperm injection cases where spermatozoa used were frozen in Cell Sleepers. Cell Sleepers are novel devices in which individual spermatozoa can be frozen in microdroplets. The case series included five men with obstructive azoospermia, six with nonobstructive azoospermia and one with cryptozoospermia, in whom microscopic sperm retrievals from testicular sperm extraction ( TESE), micro- TESE extracts and a centrifugation procedure resulted in less than 50 spermatozoa. A total of 304 microscopically retrieved spermatozoa were frozen in 20 Cell Sleepers using a rapid manual cryopreservation method. A total of 179 mature oocytes were injected with recovered thawed spermatozoa, resulting in a fertilisation rate of 65.9% (118 of 179), with no total fertilisation failures. In 10 cases, an embryo transfer was performed, three on day 3 and seven on day 5, resulting in a per cycle pregnancy rate of 58.3% (seven of 12). Four of the pregnancies have progressed past 20 gestation weeks. The recovery and use of spermatozoa that were frozen in Cell Sleepers was uncomplicated and effective and eliminated the need to perform any microscopic sperm retrieval procedures on the day of oocyte collection. Modification of the routine sperm cryopreservation methodology to include the use of Cell Sleepers increases the range of sperm samples that can be effectively cryopreserved, to include men with severe male factor fertility. [ABSTRACT FROM AUTHOR]

Sturgeon spermatozoa maturation during their passage through the kidney is a prerequisite for initiation of motility. Samples of sterlet (Acipenser ruthenus) testicular sperm (TS) were matured in vitro by incubation in seminal fluid (SF) or in SF supplemented with carbonyl cyanide m-chlorophenyl hydrazone (CCCP; a respiration uncoupling agent). Sperm was diluted in activation medium (AM) containing 10 mM Tris-HCl buffer (pH 8.5) and 0.25% Pluronic, and spermatozoon motility was assessed. Samples were taken and fixed in 3 M perchloric acid at 3 points in the incubation process. Quantification of ATP, ADP, and creatine phosphate (CrP) was conducted using liquid chromatography/high-resolution mass spectrometry. We observed a significant decrease in CrP during artificial maturation of TS in SF. In contrast, ATP and ADP were not significantly affected. Addition of CCCP to SF halted maturation and led to significantly lower CrP whereas ADP significantly increased and ATP was unaffected. Dilution of matured and immature TS with AM led to a significant decrease of ATP and CrP and an increase of ADP compared with their levels before dilution, although immature TS were not motile. Energy dependency of TS maturation in sturgeon was confirmed, which suggests that mitochondrial oxidative phosphorylation is needed for maturation of sturgeon TS. [ABSTRACT FROM AUTHOR]

Male infertility is a serious problem in an increasing number of couples. We report an infertile man with non-obstructive azoospermia and karyotype 45,XY,rob(14;22). The immunofluorescence analysis of his testicular tissue using antibodies to SYCP1, SYCP3, HORMAD2, MLH1, and centromeres showed delayed synapsis of the chromosomes involved in the translocation, a varying extent of trivalent asynapsis and its association with sex chromosomes. The mean frequency of meiotic recombination per cell was within the range of normal values. Fluorescence in situ hybridization (FISH) with probes for chromosomes 14 and 22 revealed 5.83% of chromosomally abnormal testicular spermatozoa. FISH with probes for chromosomes X, Y, and 21 showed frequencies of disomic and diploid testicular spermatozoa increased when compared to ejaculated sperm of healthy donors, but comparable with published results for azoospermic patients. PGD by FISH for the translocation and aneuploidy of chromosomes X, Y, 13, 18, and 21 showed a normal chromosomal complement in one out of three analyzed embryos. A healthy carrier girl was born after the embryo transfer. This study shows the benefits of preimplantation genetic diagnosis in a case of a rare Robertsonian translocation carrier with azoospermia and a relatively low frequency of chromosomally unbalanced testicular spermatozoa. [ABSTRACT FROM AUTHOR]

The minimum sperm count and quality that must be identified during microdissection testicular sperm extraction (micro- TESE) to deem the procedure successful remains to be established. We conducted a retrospective study of 81 consecutive men with non-obstructive azoospermia who underwent a primary (first) micro- TESE between March 2007 and October 2013. Final assessment of sperm recovery [reported on the day of (intracytoplasmic sperm injection) ICSI] was recorded as (i) successful (available spermatozoa for ICSI) or (ii) unsuccessful (no spermatozoa for ICSI). The decision to perform a unilateral (with limited or complete microdissection) or bilateral micro- TESE was guided by the intra-operative identification of sperm recovery (≥5 motile or non-motile sperm) from the first testicle. Overall, sperm recovery was successful in 56% (45/81) of the men. A unilateral micro- TESE was performed in 47% (38/81) of the men (based on intra-operative identification of sperm) and in 100% (38/38) of these men, spermatozoa was found on final assessment. In 42% (16/38) of the unilateral cases, a limited microdissection was performed (owing to the rapid intra-operative identification of sperm). The remaining 43 men underwent a bilateral micro- TESE and 16% (7/43) of these men had sperm identified on final assessment. The cumulative ICSI pregnancy rates (per cycle started and per embryo transfer) were 47% (21/45) and 60% (21/35), respectively, with a mean (± SD) of 1.9 ± 1.0 embryos transferred. The data demonstrate that intra-operative assessment of sperm recovery can correctly identify those men that require a unilateral micro- TESE. Moreover, the rapid identification of sperm recovery can allow some men to undergo a limited unilateral micro- TESE and avoid the need for complete testicular microdissection. [ABSTRACT FROM AUTHOR]

Background: Extracted sperm from the testis have poor motility. Moreover, their motility changes during their journey through epidydimis. Meanwhile, they face high concentration of L-carnitin. In addition, lactate dehydrogenase C4 (LDH-C4) gene disorders has been shown to cause impaired sperm motility, leading to infertility in male mice. The aim of this study was to evaluate sperm motility and LDH-C4 enzyme activity upon L-carnitine (LC) and Pentoxifylline (PTX) administrations in mice. Methods: We extracted testicular sperm of 48 mice and divided them into three equal parts. One part was incubated with Ham's F10 medium (control), the other parts were treated with Ham's F10 containing LC and PTX with a final concentration of 1.76 mM, for 30 min at room temperature. Sperm motility was assessed according to the World Health Organization (WHO) criteria. Sperm LDH-C4 enzyme activity was measured by spectrophotometery method. Statistical analyses were performed using ANOVA and Fisher's LSD test, and a p-value less than 0.05 was considered as a statistically significant difference. Results: Sperm motility increased after 30 min of incubation in LC- and PTX-treated group (p<0.001). LC and PTX administrations showed a significant increase in the LDHC4 enzyme activity of sperm compared to that of the controls after 30 min (P=0.04 and 0.01, respectively). Conclusion: The effects of LC and PTX on motility of sperm can be explained by an increase in LDH-C4 enzyme activity that may influence male fertility status. We suggest that LC as a non-toxic antioxidant is more suitable for use in assisted reproductive technique protocols than PTX. [ABSTRACT FROM AUTHOR]

BACKGROUND Non-obstructive azoospermic (NOA) men can father children after testicular sperm extraction (TESE). Previous studies suggest that they may be at risk of producing chromosomally abnormal spermatozoa, but the number of sperm analysed per patient was usually very low. METHODS Multicolour fluorescence in situ hybridization was used for detection of chromosome 13, 15, 16, 18, 21, 22, X and Y disomy and diploidy in sperm obtained from NOA men (n = 17) and control donors (n = 10). At least 500 testicular sperm were scored in each patient to increase the precision of our study. RESULTS The mean frequency of overall disomy (2.32%) and diploidy (0.80%) found in 13 689 testicular spermatozoa of NOA patients was significantly higher than in the ejaculated sperm of normospermic control donors, disomy (0.62%) and diploidy (0.29%). A highly significant increase in frequencies of chromosome 15, Y and overall disomy (P < 0.001), and a significant increase in disomy of chromosome 13 (P = 0.002), 16 (P = 0.031) and 21 (P = 0.018), overall diploidy (P = 0.031) and diploidy caused by errors in meiosis I (P = 0.011) were observed in the NOA group. CONCLUSIONS Testicular sperm samples of NOA patients show a higher incidence of numerical chromosomal abnormalities compared with ejaculated sperm of control donors. Appropriate genetic counselling is necessary in NOA men undergoing TESE. [ABSTRACT FROM AUTHOR]