Sperm Recovery Rate Research Articles

Seminal plasma microbiomes, sperm parameters, and cryopreservation in a healthy fertile population.

Recent advances in microbiome research have revealed the presence of diverse microbial communities in human tissues previously thought to be sterile. The present study delves into the emerging field of seminal plasma microbiomics, examining the relationship between semen microbes and semen parameters and post-freezing tolerance. The study involved a cohort of healthy fertility males and microbial genome analysis using 16S rRNA to characterize the microbial diversity of seminal plasma. Microbial diversity analysis identified unique amplicon sequence variants (ASVs) and genera dominant in seminal plasma. Spearman's correlation coefficient was used to assess the relationship between flora and semen parameters. A paired t-test was used to compare the changes in microbiome expression in seminal plasma before and after cryo-resuscitation. The relevant results show that the top five phyla in terms of abundance of seminal plasma microbiome were Firmicutes, Bacteroidota, Proteobacteria, Actinobacteriota, and Campylobacterota. Spearman correlation analysis highlighted the association between specific microbial species and semen parameters, between Porphyromonas_asaccharolytica and sperm concentration. Microbial changed significantly after cryo-resuscitation, affecting taxonomic units such as Campylobacter and Muribaculaceae, and KEGG enrichment analyses, suggesting that metabolic pathways are associated with sperm freezing. Eubacterium_coprostanoligenes and Eptoniphilus_duerdenii exhibited a potential impact, while Orynebacterium_tuberculostearicum demonstrated a positive correlation with the recovery rate of progressive motile sperm. The semen of normal fertile individuals contains a microflora component that is closely related to semen quality, including the sperm's ability to withstand freezing.

Safety evaluation of single-sperm cryopreservation technique applied in intracytoplasmic sperm injection.

Intracytoplasmic sperm injection (ICSI) is a technique that directly injects a single sperm into the cytoplasm of mature oocytes. Here, we explored the safety of single-sperm cryopreservation applied in ICSI. This retrospective study enrolled 186 couples undergoing ICSI-assisted pregnancy. Subjects were allocated to the fresh sperm (group A)/single-sperm cryopreservation (group B) groups based on sperm type, with their clinical baseline/pathological data documented. We used ICSI-compliant sperm for subsequent in vitro fertilization and followed up on all subjects. The recovery rate/cryosurvival rate/sperm motility of both groups, the pregnancy/outcome of women receiving embryo transfer, and the delivery mode/neonatal-related information of women with successful deliveries were recorded. The clinical pregnancy rate, cumulative clinical pregnancy rate, abortion rate, ectopic pregnancy rate, premature delivery rate, live birth delivery rate, neonatal birth defect rate, and average birth weight were analyzed. The two groups showed no significant differences in age, body mass index, ovulation induction regimen, sex hormone [anti-Müllerian hormone (AMH)/follicle-stimulating hormone (FSH)/luteinizing hormone (LH)] levels, or oocyte retrieval cycles. The sperm recovery rate (51.72%-100.00%) and resuscitation rate (62.09% ± 16.67%) in group B were higher; the sperm motility in the two groups demonstrated no significant difference and met the ICSI requirements. Group B exhibited an increased fertilization rate, decreased abortion rate, and increased safety versus group A. Compared with fresh sperm, the application of single-sperm cryopreservation in ICSI sensibly improved the fertilization rate and reduced the abortion rate, showing higher safety.

Cushioned and high-speed centrifugation improve sperm recovery rate but affect the quality of fresh and cryopreserved feline spermatozoa

The development of endoscopic transcervical catheterization (ETC) in the queen increases the interest in handling fresh and cryopreserved feline semen. The ETC requires a small volume of the insemination dose with a high concentration, not easily reached with the actual frozen technique in this species. Centrifugation is widely used to concentrate spermatozoa for several purposes, but this process is detrimental to spermatozoa. This study verified the effects of conventional and cushioned centrifugation on fresh and cryopreserved feline spermatozoa. To this, semen was collected from 20 toms, grouped in seven pools and diluted. After dilution, the pools were divided into two aliquots, the first used for centrifugation on fresh semen, and the second, after freezing, on cryopreserved semen. Centrifugation regimens were: conventional at 500×g, conventional at 1000×g, and cushioned (iodixanol) at 1000×g. The sperm recovery rate was calculated for the three centrifugation regimens, and sperm kinematics, membrane and acrosome integrity, and plasma membrane stability on viable spermatozoa were assessed as endpoints.The data reported in this study showed that the centrifugation at 500×g resulted in negligible effects on both fresh and cryopreserved spermatozoa, but the lower recovery rate (62.4 ± 3.1 % and 60.2 ± 1.6 %, respectively) underlines the loss of a large proportion of spermatozoa, unfavourable in a species with small total sperm ejaculated. On the other hand, the centrifugation at 1000×g improved the recovery rate (86.9 ± 4.3 % and 89.8 ± 2.4 % in fresh and cryopreserved samples, respectively), but was more deleterious for feline spermatozoa, especially in cryopreserved samples (i.e. total motility of 40.7 ± 5.4 % compared with 57.2 ± 9.8 % in cryopreserved uncentrifuged samples, P < 0.05), resulting in artificial insemination doses of lower quality. The recovery rate in cushioned centrifugation appeared less efficient, likely due to the small volume of feline samples, which makes difficult the separation of sperm pellet and cushioned fluid. Interestingly, in cryopreserved samples centrifuged at 1000×g the number of viable spermatozoa with membrane destabilization (31.3 ± 3.2 %) was greater than uncentrifuged (4.1 ± 0.7 %, P < 0.05) and those centrifuged at 500×g (9.8 ± 1.3 %, P < 0.05), suggesting modifications induced by the cryopreservation amplifies centrifugation sublethal damage on feline spermatozoa. Cushioned centrifugation on cryopreserved samples showed kinematics similar to uncentrifuged samples, but higher viable spermatozoa with membrane destabilization (37.4 ± 3.4 % vs 4.1 ± 0.7 %; P < 0.05). In felines, g-force is crucial for sperm recovery rate during centrifugation, with better results at 1000×g; on the other hand, greater g-forces could have a significant impact on the quality of feline insemination dose, especially in cryopreserved samples.

#353 : Microfluidic Preparation of Spermatozoa: A Potential Approach in Assisted Reproductive Technology

Background and Aims: While density gradient centrifugation (DGC) is a conventional method in sperm preparation, this method can induce reactive oxygen species (ROS) which cause sperm dysfunction, sperm DNA damage, a recent advancement in sperm processing using microfluidic system is believed to reduce damage and to select the good quality sperm. The aim of this study was to evaluate the effectiveness of sperm preparation between DGC method and microfluidic system. Method: 30 semen samples were separated into 2 equal small samples, each sample was processed by DGC methods (control) and microfluidic (intervention) respectively. Pre- and post processing parameters was evaluated according to WHO-2010 and the DNA fragmentation index (DFI) was determined by flow cytometry technique. Results: The result showed that DFI values from the microfluidic preparation (0.67 ± 0.53%) was significantly lower than that of the DCG group (1.18 ± 13.73%). In particular, for semen samples with high DFI (&gt;15%), the microfluidic system showed a noticeable reduction in DFI compared to the results from the DGC method. The progressive motile sperm recovery rate was higher in the intervention group (39.75 ± 19.20 vs 32.14 ± 18.14). Our data also showed that the vitality and mobility of sperm in the microfluidic group was higher than those of the DGC group (97.9 ± 2.21 vs 96.56 ± 4.11%, 96.36 ± 2.60 vs 95.06 ± 2.35%, respectively). However, there was no difference in sperm progressive motility and sperm morphology between two groups. Conclusions: There were significant improvement in sperm DFI and progressive motility in the microfluidic system. With this result, microfluidic system is a potential approach to reduce DNA fragmentation for ICSI procedures.

ICSI outcome after microdissection testicular sperm extraction, testicular sperm aspiration and ejaculated sperm

We conducted a case-controlled single-center cohort study to evaluate the intracytoplasmic sperm injection (ICSI) outcome in severe male infertility with different methods of sperm obtention. The data was compiled from a tertiary university hospital. The micro-TESE procedures were performed from 2008 to 2023, with a sperm recovery rate (SRR) of 45 %. The ICSI treatments were carried out between 2011 and 2023. The aim of the study was to compare the ICSI outcome using sperm obtained by microdissection testicular extraction (micro-TESE), testicular sperm aspiration (TESA), and ejaculated sperm with sperm concentration less than 15 million per milliliter. We included a total of 462 ICSI cycles, of which 340 ICSIs with ejaculated sperm of men with oligozoospermia, with or without asthenozoospermia or teratozoospermia (OAT group), 51 ICSIs with TESA sperm of men with obstructive azoospermia (OA, TESA group), and 71 ICSIs with micro-TESE sperm of men with non-obstructive azoospermia (NOA, micro-TESE group). The patient characteristics, fertilization rate, pregnancy rate, and pregnancy outcome data were similar between the groups. The fertilization rates were 66.0 % in the OAT group, 68.3 % in the TESA group and 62.8 % in the micro-TESE group and live birth rate per embryo transfer were 23.7 %, 28.9 %, and 25.0 %, respectively, without statistical difference. The obstetrical outcome was similar in all the groups. The overall clinical results in all ICSI cycles performed for treating severe male factor infertility were similar, independent of the method of collection of spermatozoa. The results also confirm the efficacy of micro-TESE in the treatment of severe male factor infertility.

Leydig cell metabolic disorder act as a new mechanism affecting for focal spermatogenesis in Klinefelter syndrome patients: a real world cross-sectional study base on the age.

Klinefelter's syndrome (KS) was once considered infertile due to congenital chromosomal abnormalities, but the presence of focal spermatozoa changed this. The key to predict and promote spermatogenesis is to find targets that regulate focal spermatogenesis. To explore the trend of fertility changes in KS patients at different ages and identify potential therapeutic targets. Bibliometric analysis was used to collect clinical research data on KS from the Web of Science Core Collection (WoSCC) from 1992 to 2022.A cross-sectional study was conducted on 75 KS patients who underwent microscopic testicular sperm extraction (mTESE) from 2017 to 2022 in the real world. The reproductive hormones, testicular histopathology, androgen receptors, insulin-like factor 3 (INSL3) receptors and sperm recovery rate (SRR) were analyzed. Male infertility, dysplasia, Sertoli cells, Leydig cells, testosterone and spermatogenesis were the research focuses related to KS. Luteinizing hormone (LH), testosterone, and INSL3 were evaluation indicators of Leydig cell function that fluctuate with age. Testosterone and LH peaked at ages 13-19 and 30-45, while INSL3 only peaked at ages 13-19. 27 patients (27/75) recovered sperm through mTESE and experienced SRR peaks at the ages of 20, 28, 34, and 37. The SRR of fibrosis patients was 46.15%, fatty degeneration was 7.14%, and melanosis was 40.00%. The INSL3 and androgen receptors were highly expressed and roughly balanced in focal spermatogenesis. Abnormal metabolism of Leydig cells led to imbalanced expression of INSL3 and androgen receptors, which might be a potential target for spermatogenesis in KS.

Processing stallion semen for on‐farm use and cooled transport

SummarySimple dilution of semen is the oldest method to process semen for artificial insemination with either fresh or cooled semen. This was developed prior to the optimisation of centrifugation techniques for semen processing through a combination of trial and error and research. Simple dilution is a faster method to process semen, is easy to use and works in many circumstances. For on‐farm semen use a 1:1 (v/v) semen to extender ratio is used. A variety of other techniques have been developed, such as cushion centrifugation, density gradient centrifugation and sperm filtration. Cushion centrifugation is used to concentrate the sperm and allows higher centrifugation speeds than centrifugation alone. This allows for improved recovery rates of sperm. Density gradient centrifugation has been used to select for morphologically normal sperm but suffers from low recovery rates. Processing sperm with this technique can result in improved motility and improved DNA integrity of sperm. Sperm filtration is another method to concentrate sperm without the use of centrifugation; however, it has lower recovery rates compared to cushion centrifugation. This review will discuss these techniques in the application of processing equine semen for either fresh or cooled‐shipped use.

Effect of ascorbic acid on metabolic status, lipid peroxidation, antioxidant activity and quality of frozen Indian red jungle fowl (Gallus gallus murghi) semen.

The Indian red jungle fowl population is decreasing in its natural habitat. Its conservation through semen cryopreservation with sufficient live sperm recovery rate is requisite where ascorbic acid could play significant role to mitigate cryo-incited injuries. The objective was to elucidate the effect of ascorbic acid on freezability of Indian red jungle fowl sperm. Pooled semen was aliquoted and diluted (1:5) with red fowl extender having ascorbic acid: 0.0 (control), 1.0, 2.0 and 4.0 mM. Diluted samples were cryopreserved and semen quality was assessed at post-dilution, cooling, equilibration and freeze-thawing stages. Sperm metabolic status, antioxidant potential and lipid peroxidation were studied at post-dilution and freeze-thawing. Sperm motility did not differ (p > .05) in experimental extenders and control at post-dilution and cooling; however, it was recorded higher (p < .05) with ascorbic acid at 2.0 mM compared with other levels at post-equilibration and post-thawing stage. Sperm viability, plasma membrane and acrosome intactness were recorded higher (p < .05) with 2.0 mM ascorbic acid compared with other concentrations of ascorbic acid at all stages of cryopreservation. Sperm metabolic status and antioxidant potential were recorded higher (p < .05), while lipid peroxidation was recorded lowest (p < .05) with 2.0 mM ascorbic acid compared with 1.0, 4.0 mM and control. In conclusion, ascorbic acid at 2.0 mM in red fowl extender improves quality, metabolic status and antioxidant potential of frozen Indian red jungle fowl semen through ameliorating lipid peroxidation.

Assessing mail-in cryopreservation for fertility preservation.

1525 Background: Over 90% of oncologists agree that sperm banking should be offered to all men at risk of subfertility associated with cancer treatment, but fewer than 50% bring up the topic. Barriers cited include lack of time for the discussion, perceived high cost, and lack of convenient facilities. The advent of mail-in semen analysis (SA) testing and cryopreservation may improve access to this service via lower patient cost, decreased required oncologist logistical support, and home rather than local lab collection. That said, average sperm recovery rate after sample thaw is ~50% with immediate banking, and sperm recovery rate after delayed cryopreservation with a mail-in approach remains unknown. Methods: This is a prospective preliminary study assessing freeze-thaw total motile count at 3 time points of cryopreservation: upon receipt of sample, at 24 hours, and at 48 hours post collection. We collected 39 fresh semen samples from 13 healthy adult volunteers. Samples were diluted with 5mL of Fellow Health preservation solution and analyzed (World Health Organization guidelines V5). Samples were divided into 3 aliquots: Aliquot #1, T0 was immediately frozen with equal part cryoprotectant. Aliquot #2, T24 and aliquot #3, T48 were left at room temperature for 24 and 48 hrs respectively, analyzed, and then frozen. Aliquots were then thawed on a heat block at 37C for 10 minutes and analyzed. Results: Baseline SA values were: volume 3.3ml +/- 1.3, concentration 138.3 +/-57.4 x 106, motility 49% +/-6%; Total Motile Count (TMC) 213 +/- 110 million. TMC was the primary endpoint as measured by baseline semen volume, concentration, and motility at 3 time points (T0, T24, T48). The average loss from freeze/thaw in TMC at T0 was 45%, at T24 was 55% relative to T24 pre-freeze, and at T48 was 59% relative to T48 pre-freeze (p = 0.013). On average, samples with starting TMC between 50-100 million had enough motile sperm for six intrauterine insemination (IUI) cycles when frozen and thawed at T0, two at T24, and two at T48. Samples with starting TMC between 100-200 million had enough motile sperm for 16 IUI cycles when frozen and thawed at T0, eight at T24, and four at T48. Samples with starting TMC &gt; 200 million had enough motile sperm for 42 IUI cycles when frozen and thawed at T0, 21 at T24, and nine at T48. All samples had ample viable cells ( &gt; 50,000) at all three time points for in vitro fertilization (IVF). Conclusions: Mail-in cryopreservation using our improved methodology may increase access to this service for oncological patients while still facilitating reasonable assisted reproductive technology care relative to in-person cryopreservation.

Role of Repeated Sperm Extraction by Enzymatic and Mechanical Mincing Techniques in Azoospemic Patients

Azoospermic patients have the option of undergoing in vitro fertilization procedures, specifically intracytoplasmic sperm injection (ICSI), for which sperm is retrieved from the patient's own testicles and then injected directly into the oocyte. The aim of the study: evaluate the role of repeated sperm extraction by enzymatic and mechanical mincing techniques in azoospemic patients. A total of 32 males with non-obstructive azoospermia participated in the research. Testicular sperm extraction (TESE) was performed under spinal or general anesthesia after a comprehensive history, physical examination, and investigations. After that, the biopsies underwent through a series of mincing processes, both mechanical and enzymatic. Patients gave information including age, body mass index and previous biopsy attempts. In both enzymatic and mechanical methods there was no significant higher sperm recovery in patients with history of previous testicular biopsies (66.7% vs. 38.5%; p=0.209) and (66.7% vs. 30.8%; p=0.102) respectively. Patients who had undergone prior testicular biopsies had marginally higher sperm recovery rates using both enzymatic and mechanical techniques

Infertility treatment for patients having a microdeletion of azoospermic factor (AZF).

In genetic causes of male infertility, Y chromosome microdeletions are the second most common after Klinefelter's syndrome. Although sperm recovery rate is relatively high for subjects with azoospermic factor (AZF) c chromosome microdeletion, intracytoplasmic sperm injection (ICSI) results using retrieved sperm has been reported to be poor. We retrospectively examined the infertility treatment for subjects with AZF microdeletion. From October 2017 to September 2020, chromosomal examination of 67 azoospermic subjects and 12 cryptozoospermia were performed. Of these, twenty-three subjects (29.1%) had AZF microdeletion. Twelve subjects with AZFc microdeletion and one subtype with unknown classification (Ym-9; P3 deletion) received sperm retrieval surgery due to azoospermia. Two subjects obtained motile sperm by microscopic epididymal sperm aspiration (MESA) and four subjects by microscopic testicular sperm extraction (micro-TESE). Pregnancy and healthy delivery were achieved in 6 of 14 subject (42.9%; including one twin) using ICSI. This was comparable with previous reports. Since there were two cases of obstructive azoospermia, we employed MESA to avoid testicular damage. Following observation of the testis and epididymis under operative microscope, a decision was made to perform sperm retrieval surgery to avoid unnecessary testicular damage. Furthermore, since AZFc microdeletion is passed to the next generation, long term follow-up is necessary.

Defining an Optimal Range of Centrifugation Parameters for Canine Semen Processing.

Our objective was to determine a clinically relevant range of centrifugation parameters for processing canine semen. We hypothesized that higher gravitational (g) force and longer time of centrifugation would result in improved spermatozoa recovery rate (RR) but poorer semen quality. Cooled storage under standard shipping conditions was used as a stressor to evaluate long-term treatment effects. Individual ejaculates collected from 14 healthy dogs were split into six treatment groups (400 g, 720 g, and 900 g for 5 or 10 min). Sperm RR (%) was calculated post-centrifugation, and plasma membrane integrity (%, Nucleocounter® SP-100™), total and progressive motility (%, subjective and computer-assisted sperm analysis), and morphology (%, eosin-nigrosin staining) were assessed on initial raw semen (T0), post-centrifugation (T1), and 24 h (T2) and 48 h (T3) after cooling. Sperm losses were minimal, and RRs were similar across treatment groups (median >98%, p ≥ 0.062). Spermatozoa membrane integrity was not different between centrifugation groups at any time point (p ≥ 0.38) but declined significantly during cooling (T1 vs. T2/T3, p ≤ 0.001). Similarly, total and progressive motility did not differ across treatments but declined in all groups from T1 to T3 (p ≤ 0.02). In conclusion, our study showed that centrifugation within a range of 400 g-900 g for 5-10 min is appropriate for processing canine semen.

A novel, simplified method to prepare and preserve freeze-dried mouse sperm in plastic microtubes.

Although freeze-drying sperm can save space, reduce maintenance costs, and facilitate the transportation of genetic samples, the current method requires breakable, custom-made, and expensive glass ampoules. In the present study, we developed a simple and economical method for collecting freeze-dried (FD) sperm using commercially available plastic microtubes. Mouse epididymal sperm suspensions were placed in 1.5 ml polypropylene tubes, frozen in liquid nitrogen, and dried in an acrylic freeze-drying chamber, after which they were closed under a vacuum. The drying duration did not differ between the microtube and glass ampoule methods (control); however, the sperm recovery rate was higher using the microtube method, and the physical damage to the sperm after rehydration was also reduced. Intracytoplasmic sperm injection (ICSI) using FD sperm stored in microtubes at -30°C yielded healthy offspring without reducing the success rate, even after 9 months of storage. Air infiltration into all microtubes stored at room temperature (RT) within 2 weeks of storage caused a drastic decrease in the fertilization rate of FD sperm; underwater storage did not prevent air infiltration. RT storage of FD sperm in microtubes for 1 week resulted in healthy offspring after ICSI (5-18%), but the addition of silica gel or CaCl2 did not improve the success rate. Our novel microtube method is currently the simplest and most effective method for treating FD sperm, contributing to the development of alternative low-cost approaches for preserving and transporting genetic resources.

In vitro approach points to a chemotactic effect of melatonin on ram spermatozoa

Sperm orientation mechanisms, such as chemotaxis, are essential for the sperm to reach the oocyte and fertilize it. Melatonin is secreted by the cumulus cells and is also present in the follicular fluid in mammals. The presence of membrane receptors for melatonin in ram spermatozoa, and its proven involvement in the sperm functionality, may suggest a possible role in the guided movement towards the oocyte. Hence, the objective of the present work is to study the in vitro potential chemotactic action of melatonin on ram spermatozoa, analysing the influence of the season (breeding and non-breeding) and the sperm capacitation state. The first experimental approach consisted in the inclusion of melatonin in the upper layer of a swim-up selection method. During the non-breeding season, the presence of melatonin at 100 pM and 1 μM concentrations significantly increased the cell recovery rate, and induced changes in the sperm location of the MT2 melatonin receptor, compared with the standard swim-up. Moreover, the selected sperm population with 100 pM melatonin presented a higher percentage of capacitated spermatozoa. The greater recovery rate obtained with melatonin could be due to the stimulation of sperm movement in random directions, i.e., a chemokinetic effect, or due to a guided movement (chemotaxis) towards the gradient of the melatonin. To elucidate this issue, together with the study of the influence of the sperm capacitation status, we performed a second experimental approach which consisted in the use of chemotaxis chambers and an open-source software (Open-CASA) that analyses the sperm trajectories towards the hormone gradient and calculates a chemotaxis index (SL index). There was a significant difference between the SL index in the presence of 1 μM melatonin and the control without hormone. This effect was only observed in capacitated spermatozoa with cAMP-elevating agents (Cap-CK samples) obtained during the non-breeding season. These results would point to an in vitro chemotactic effect of melatonin on ram spermatozoa, although chemokinesis cannot be ruled out. Nonetheless, the inclusion of this hormone in the swim-up procedure could enhance the sperm recovery rate.

An evaluation of the population characteristics, semen quality, and utilization status of autologous sperm cryopreservation and fertility preservation in for 662 patients: a 6-year monocentric retrospective study

BackgroundSperm cryopreservation is an effective method of fertility preservation for disease-related and social sperm freezing. In total, 662 subjects (range: 15–65 years-of-age; mean: 33.49 ± 8.79 years-of-age) were included in this study to investigate the population characteristics, semen quality, and usage of autologous sperm preservation patients in Beijing. Of these, 351 were cancer patients (53.02%, 31.14 ± 7.32 years-of-age) and 311 were non-cancer patients (46.98%, 36.14 ± 9.54 years-of-age).ResultsWe found that the number of preservation cases increased steadily from 2015 to 2019; 89.73% of these had a bachelor's degree or above; 54.83%, 41.54%, and 3.63% were single, married, and divorced, respectively. The cases of cancers and oligozoospermia accounted for 71.30% of all patients; therefore, most patients required fertility preservation due to disease. The cancer group had a significantly lower sperm concentration, rate of progressive sperm after the frozen-thawed test, total progressive motility sperm count after the frozen-thawed test, and recovery rate of progressive motile sperm (RRPM) than the non-cancer group (all P < 0.05). Sperm count-related parameters were significantly affected by testicular cancer, while sperm motility-related parameters and RRPM were significantly affected by leukemia. The utilization rate of preserved sperm was 6.34% after 6 to 78 months of follow-up. In terms of fresh or frozen embryo transfer, the clinical pregnancy rate was 56.76% or 50.00%, and the live birth rate was 24.32% or 21.43%, respectively.ConclusionThe need for autologous sperm preservation was dominated by patients with diseases, followed by the need for social sperm freezing. Tumors had a major negative impact on semen quality, and the usage rates of stored semen were at lower level compared to the number of sperm cryopreservation. Medical staff and patients should pay attention to both cognition-action consistency and cost-effectiveness in fertility preservation.

A novel solution for freezing individual spermatozoa using a right angular cryopiece embedded in a grooved petri dish.

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.

Daya Tahan Tudung Akrosom dan Membran Plasma Spermatozoa Sapi Bali yang Dipaparkan pada Suhu Ruang

This study aimed to determine the effect of storage at room temperature on the durability of the acrosome cap and plasma membrane of Bali cattle spermatozoa. This research was conducted at the Reproduction Laboratory of the Faculty of Animal Husbandry, Jambi University. The experiment was carried out using a completely randomized design with 4 treatments and 6 replications. Storage treatment at room temperature consisted of T0: no storage (control),T1: storage for 2 hours, T2: storage for 4 hours, T3: storage for 6 hours. The data obtained from each variable analyzed with variance. The treatment had a significant effect, so Duncan's further test was carried out. The process of calculating the data using the SPSS program. From the results of this study, it was found that storage of spermatozoa at room temperature reduced the quality of Bali cattle's spermatozoa against the acrosome hood, plasma membrane, and spermatozoa recovery rate (P&lt;0.05). At storage for up to 4 hours, both 44,33% acrosomal cap and 52.83% recovery rate were still eligible for insemination. In plasma membrane storage at room temperature ranging from 0 hours 56.60% to 6 hours 35.83% showed a fairly low average, which was below 60% as the minimum requirement for insemination

DEVELOPMENT OF A MACHINE LEARNING APPLICATION FOR INTRAOPERATIVE OBJECT DETECTION OF POSITIVE SEMINIFEROUS TUBULES IN MICRODISSECTION TESTICULAR SPERM EXTRACTION FOR NONOBSTRUCTIVE AZOOSPERMIA

Approximately 1 in 100 men in the general population have azoospermia, and 80% of them have nonobstructive azoospermia (NOA). For couples with NOA who wish for a baby, the only effective treatment options are sperm retrieval by microdissection TESE (micro-TESE) and subsequent ICSI. However, the current sperm recovery rate is generally only 30-40%. Favorable seminiferous tubules that contain sperm are generally white and thicker in diameter than other tubules, and the success of sperm retrieval is largely dependent on the surgeon's attentiveness and observational ability.

IMPACT OF MICROFLUID SPERM SORTING ON EMBRYO PLOIDY RATES IN COUPLES WITH HIGH SPERM DNA FRAGMENTATION?

Increased sperm DNA fragmentation (SDF) is one of the leading causes of male infertility. Conventional methods for sperm preparation have induced questioning of sperm recovery rates. The microfluidic sperm sorting (MSS) technique selects highly motile sperm with lower levels of SDF compared to density-gradient centrifugation (DGC) technique. This study aimed to compare MSS to DGC technique in couples with repeated implantation failure (RIF) and high SDF referred to a PGT-A (preimplantation genetic testing for aneuploidies) cycle.

Sperm selection techniques in cattle: Microfilter device versus conventional methods

Microfluidics and microfilter devices have been developed to mimic the characteristics of the female reproductive tract, minimizing the risk of sperm damage. This study aimed to compare the use of a microfilter device versus conventional methods for sperm selection used in in vitro fertilization (IVF). For selecting spermatozoa, the pooled samples were processed in a microfilter device, swim-up and mini-Percoll gradient. Kinematic and morphometric parameters, vitality and DNA damage were analysed before and after sperm selection. After selection, 10,000 motile spermatozoa per oocyte were used in IVF drops. Embryos were assessed at three (cleavage rate) and seven (blastocyst rate) days post-IVF. Results of sperm kinematic parameters including average path velocity, velocity straight line, curvilinear velocity, linearity, lateral head displacement with the microfilter device were superior to density gradient (p < 0.05), but similar to swim-up method. Likewise, sperm DNA damage was significantly reduced using the microfilter device and swim-up method. Regarding the total sperm recovery rate post selection, results with the microfilter device (17.64%) and mini-Percoll gradient (18.27%) were higher than with swim-up method (6.52%). However, the cleavage and blastocyst rates were the lowest using the microfilter device. In conclusion, sperm selection using the microfilter device and swim-up method can improve kinematic parameters, although the mini Percoll gradient was the most efficient method for embryo production.