Sperm Preservation Research Articles

Can We Cryopreserve the Sperm of COVID-19 Patients During the Pandemic?

An extreme strain has been placed on healthcare facilities in the COVID-19 era. Initial stage of the pandemic, national and international societies for reproductive medicine suggested the suspension of new IVF treatments and non-essential cryopreservation of gametes. Accordingly, the demands of cryopreservation of semen with COVID-19 patients also was suspended by some of cryobanks to protect staff and patients from unnecessary viral exposure at the acute stage. However, the pandemic may stay with us longer than expected. In addition, there will be some male COVID-19 patients with cancer or critically illness who needs to cryopreserve their semen before medical treatments, otherwise they might loss the chance of getting their own offspring. In this document, we summarize available evidence to deepen and expand awareness of feasibility of sperm cryopreservation and propose some suggestions to help cryobanks carry out sperm preservation procedure for COVID-19 male patients.

Challenges of sperm cryopreservation in transferring heat adaptation of corals across ocean basins.

Reef-building corals live very close to their upper thermal limits and their persistence is imperiled by a rapidly warming climate. Human interventions may be used to increase the thermal limits of sensitive corals by cross-breeding with heat-adapted populations. However, the scope of breeding interventions is constrained by regional variation in the annual reproductive cycle of corals. Here we use cryopreservation technology to overcome this barrier and cross-breed conspecific coral populations across ocean basins for the first time. During regional spawning events, sperm samples were cryopreserved from populations of the widespread Indo-Pacific coral, Platygyra daedalea, from the southern Persian Gulf (maximum daily sea surface temperature of 36 °C), the Oman Sea (33 °C), and the central Great Barrier Reef (30 °C). These sperm samples were thawed during a later spawning event to test their ability to fertilize freshly spawned eggs of P. daedalea colonies from the central Great Barrier Reef. Average fertilization success for the Persian Gulf (9%) and Oman Sea (6%) sperm were 1.4–2.5 times lower than those for the native cryopreserved sperm from Great Barrier Reef (13–15%), potentially due to lower sperm quality of the Middle Eastern sperm and/or reproductive incompatibility between these distant populations. Overall, fertilization success with cryopreserved sperm was low compared with fresh sperm (>80%), likely due to the low motility of thawed sperm (≤5%, reduced from 50% to >90% in fresh sperm). To evaluate whether cross-bred offspring had enhanced thermal tolerance, the survival of larvae sired by Persian Gulf cryopreserved sperm, Great Barrier Reef cryopreserved sperm, and Great Barrier Reef fresh sperm was monitored for six days at ambient (27 °C) and elevated (33 °C) temperature. Against expectations of thermal tolerance enhancement, survival of larvae sired by Persian Gulf cryopreserved sperm was 2.6 times lower than larvae sired by Great Barrier Reef fresh sperm at 33 °C (27% versus 71%), but did not differ at 27 °C (77% versus 84%). This lack of enhanced thermal tolerance was unlikely due to outbreeding depression as survival was equally poor in larvae sired by Great Barrier Reef cryopreserved sperm. Rather, follow-up tests showed that cryoprotectant exposure during fertilization (0.1% DMSO) has a negative effect on the survival of P. daedalea larvae which is exacerbated at elevated temperature. Collectively, our findings highlight challenges of breeding corals for enhanced thermal tolerance using cryopreserved sperm, which may be overcome by methodological advances in the collection and preservation of high-quality motile sperm and minimizing the exposure time of eggs to cryoprotectants.

Motility and oxidative stress of common carp Cyprinus carpio sperm during short-term storage

Short-term storage of semen is a simple and inexpensive procedure to deal with logistics of large-scale hatchery operations but can lead to oxidative stress and a significant decrease in sperm motility and velocity. To better understand the mechanisms responsible for the association of poor sperm quality with oxidative stress, in the present study we investigated the effect of refrigerated storage for 0, 24, 48, 72, and 144 h on sperm motility and curvilinear velocity and oxidant/antioxidant balance of common carp Cyprinus carpio. Percentage of motile sperm was significantly (p < 0.05) reduced stored for 72 h storage compared to that of fresh sperm. Sperm stored 144 h showed < 40% motility with an average velocity of 91.12 ± 10.4 µm s−1. A time-dependent increase in the level of the oxidative stress indices lipid peroxidation and carbonyl derivatives of proteins was observed. Increase (p > 0.05) in total superoxide dismutase was detected after 48 h, and glutathione reductase and glutathione peroxidase activity demonstrated a significant increase after 72 h. These results provide an additional tool for the development and improvement of short-term sperm preservation procedures commonly applied in aquaculture.

MP28-17 CHARACTERIZING THE CHALLENGES TRANSFEMININE INDIVIDUALS FACE IN SEEKING UROLOGICAL CARE

You have accessJournal of UrologyCME1 May 2022MP28-17 CHARACTERIZING THE CHALLENGES TRANSFEMININE INDIVIDUALS FACE IN SEEKING UROLOGICAL CARE Andrew Denisenko, Andrew Salib, Jessica Bulafka, Sabina Spigner, Vishal Swaminathan, Peyton Stauffer, Cassra B. Clark, Andrew Shumaker, Rosemary Frasso, and Paul H. Chung Andrew DenisenkoAndrew Denisenko More articles by this author , Andrew SalibAndrew Salib More articles by this author , Jessica BulafkaJessica Bulafka More articles by this author , Sabina SpignerSabina Spigner More articles by this author , Vishal SwaminathanVishal Swaminathan More articles by this author , Peyton StaufferPeyton Stauffer More articles by this author , Cassra B. ClarkCassra B. Clark More articles by this author , Andrew ShumakerAndrew Shumaker More articles by this author , Rosemary FrassoRosemary Frasso More articles by this author , and Paul H. ChungPaul H. Chung More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002571.17AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Medical providers will encounter transgender individuals in their practice, and it is important that they understand the unique needs and backgrounds of this population. We sought to characterize the challenges transfeminine individuals face in seeking urological care through a developed questionnaire. METHODS: IRB approval was obtained to develop (with feedback from the transgender community) and administer a 58-item questionnaire assessing urinary and sexual health. Participants were recruited from the local community and the Philadelphia Trans Wellness Conference and provided a $20 gift card for their time. We assessed demographic factors, therapies, surgeries, complications, and urologic care experiences of these participants and compared vaginoplasty and non-vaginoplasty patients. RESULTS: 53 transfeminine individuals (11 vaginoplasty, 10 orchiectomy, 32 no procedure) with mean age of 40 (range 21-80) completed the questionnaire. Of the 12 respondents who met age-based criteria for prostate cancer screening, 8 underwent screening, 4 did not undergo screening, and 2 were not offered screening. 30 (56%) patients were not offered sperm preservation counseling. Vaginoplasty complications were reported by 6 respondents (54%) with 3 reports of wound separation, numbness, or inadequate vaginal depth, and single occurrences of abnormal scarring, vaginal stenosis, meatal stenosis, rectovaginal fistula, and chronic pain (n=11). We noted that those who had undergone vaginoplasty were significantly more likely to see a provider for urinary complaints (p=0.042), with 3 (27%) of these patients electing to see a urologist. Of the 14 patients from the entire cohort who were not willing to see a urologist, 29% cited discomfort with discussing urinary or sexual issues, while 7% cited fear of being misgendered. Across both groups, 30 individuals (57%) reported that they experienced a lack of knowledge about transgender health needs from members of a urology practice. CONCLUSIONS: Our findings illustrate the challenges transgender individuals face in obtaining care. We noted that participants who had vaginoplasty were more likely to see a urologist, which may reflect greater familiarity with the medical system. Many patients are reluctant to seek care due to discomfort or fear, and a majority experienced lack of knowledge about their health needs. This highlights a critical need for provider education on transgender issues in order to provide a safe and inclusive environment to better care for these patients. Source of Funding: World Professional Association for Transgender Health © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e467 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Andrew Denisenko More articles by this author Andrew Salib More articles by this author Jessica Bulafka More articles by this author Sabina Spigner More articles by this author Vishal Swaminathan More articles by this author Peyton Stauffer More articles by this author Cassra B. Clark More articles by this author Andrew Shumaker More articles by this author Rosemary Frasso More articles by this author Paul H. Chung More articles by this author Expand All Advertisement PDF DownloadLoading ...

Fertility Rate and Assessment of the Cytoprotective Capacity of Various Types of Holothuroidea Extracts on Spermatozoa.

For years, compounds of natural origin have been the subject of extensive biomedical research due to very interesting, new ingredients potentially useful for various pharmaceutical, medical and industrial applications. The therapeutic properties and healing benefits of sea cucumbers may result from the presence of numerous, biologically active ingredients. Sperm subjected to processing and subsequent storage at low temperatures experience a number of damage, including the loss of the integrity of the cytoplasmic membrane, DNA and acrosome defragmentation. Therefore, the aim of this experiment was to investigate the cytoprotective potential of sea cucumber extract against cryopreserved sperm and semen fertility rate. Commercially available sea cucumber extract was taken from the cellulose shell, then 790 mg of powder was weighed out and placed in 3 glass tubes containing, respectively: 10 mL of water-glycerin solution (WG), water-ethanol (EC), glycerin-ethanol (GE), glycerin-DMSO (DG). Tubes were mixed with vortex for 3 min, then placed in a water bath and incubated for 16 h at 40 °C. Six simmental bulls, 3 years old, of known health status were used for the experiment. Semen was collected from each male once a week (for 18 weeks) using an artificial vagina. After an initial assessment of semen quality, the ejaculates were pooled to eliminate individual differences between males, then diluted to a final concentration of 80 × 106 sperm/mL with a commercial extender (Optixcell, IMV, L’Aigle, France) and divided into 16 equal samples. Control (C) without additive, the test samples contained 2, 4, 6, 8 and 10 µL WG, 2, 4, 6, 8 and 10 µL WE, 2, 4, 6, 8 and 10 µL GE, 2, 4, 6, 8 and 10 µL DG. Semen was frozen/thawed and assessed for motility, viability, DNA defragmentation, mitochondrial membrane potential and acrosome integrity. It was shown a positive effect of water-glycerin (WG) and glycerine-ethanol (GE) extracts on the efficiency of sperm preservation at low temperatures. Established that, depending on the type of prepared extract, the sea cucumber can have both cytoprotective (WG, GE, WE) and cytotoxic (DG) effects. Moreover, too high concentrations of the extract can adversely affect the sperm in terms of parameters such as viability, motility, mitochondrial potential, and the integrity of the acrosome or DNA of cells. The present study, thanks to the use of model animals to study the cytoprotective potential of the sea cucumber extract, proves that it can be a potential candidate for use in semen cryopreservation technology to improve the efficiency of storage at low temperatures. Further research is needed to optimize the composition of individual types of extracts and their effect on sperm. The highest effectiveness of female fertilization was observed when doses from GE groups (2 and 4) were used for insemination. The results of this analysis prove that the addition of the tested extract may improve the fertilization efficiency.

Exploring the causes of semen quality changes post-bariatric surgery: a focus on endocrine-disrupting chemicals.

Rapid weight loss promoted by bariatric surgery (BS) can release accumulated lipophilic endocrine-disrupting chemicals (EDCs), making these chemicals systemically available. Men typically have a higher EDC body burden and lose more weight post-BS than women, which may put male BS patients at high risk for testicular toxicity. In this review, we analyze the impacts of BS on semen parameters with a particular focus on the potential effects of EDCs. After BS, serum EDC concentrations progressively increase; and there is evidence that semen parameters deteriorate after BS. Although elevated serum EDC concentrations are associated with inferior sperm parameters, links between semen parameters and EDCs post-BS have not been studied. Understanding these potential associations requires adequately powered studies, particularly within prospective longitudinal cohorts with long-term follow-up for sperm parameters, nutritional status, sex-hormones levels and serum EDC concentrations. Studying BS patients prospectively provides the important opportunity to evaluate dose-response effects of EDC serum concentrations on sperm quality and function. Research is also needed to identify critical chemical exposure periods post-BS to inform reproductive decisions, including consideration of sperm preservation before surgery.

Effect of Nelumbo nucifera Petals Extract on Antioxidant Activity and Sperm Quality in Charolais Cattle Sperm Induced by Mancozeb.

The white Nelumbo nucifera petals aqueous extraction (NAE) was prominent in phytochemical content, antioxidant activity, and enhanced rat sperm viability induced by FeSO4, a heavy metal. Mancozeb (MZ) contains heavy metals and is widely used for fungal control in agriculture and industry. It induces oxidative stress and causes of spermatogenesis and reproductive organs’ abnormalities in both humans and animals. The aims of the present study were to investigate the effects of white Nelumbo nucifera petals aqueous extraction (WNAE) on sperm quality in cattle sperm induced by MZ. Moreover, this study investigated phytochemical compounds by liquid chromatography-mass spectrometry. A protein profile related to sperm quality with SDS-page and sperm energy preservation for each treatment was determined. The results found nine phytochemical compounds, in which quercetin-3-O-arabinoglycoside was a major flavonoid that was found in the WNAE. MZ induced free radicals in cells, leading to LPO and protein oxidation, while decreasing sperm motility, sperm viability, acrosome integrity, and normal sperm morphology. The cattle sperm found four proteins related to sperm quality including MWs of 17, 31, 34, and 55 kDa. The WNAE effectively increased energy preservation, sperm motility, sperm viability, acrosome integrity, and normal sperm morphology. The WNAE enhanced sperm qualities by reducing oxidative stress. It might be suggested that WNAE has benefits for sperm preservation which may be used to guard against toxicity in animals or humans exposed to MZ contaminants.

Transcriptome analysis of inseminated sperm storage tubules throughout the duration of fertility in the domestic turkey, Meleagris gallopavo

Sperm storage tubules (SST) are specialized invaginations of the oviductal epithelium that permit avian species to store spermatozoa for extended periods of time, without compromising sperm fertilization capacity. The molecular and physiological mechanisms behind sperm storage tubule differentiation, sperm protection, and regression remain largely unknown, but most likely have potential implications for substantially improving hen fertility, sperm storage, and semen cryopreservation in commercial poultry species. RNA sequencing was performed on sperm storage tubules isolated from the epithelium of the uterovaginal junction (UVJ) from hens at d 1, 7, 30, 60, and 90 postinsemination (n = 4 per timepoint). Read mapping and differential expression analysis were performed using CLC Genomics Workbench. A total of 2,340 differentially expressed genes were subjected to pathway analysis through Ingenuity Pathway Analysis (IPA). Through functional annotation of differentially expressed genes during early, peak, and late egg production, novel insights regarding the role of innate and acquired immune response to sperm, lipid synthesis and transfer, steroid hormone signalling, cytoskeletal reorganization, and regulation of ion homeostasis in SST were obtained. Additionally, potential pathways were identified that could be involved with suppressing sperm motility while sperm reside within the SST. Upstream analysis identified potential regulatory roles for 18 upstream regulators that could modulate sperm storage tubule function, including suppression of sperm motility. Understanding sperm storage tubule function throughout the laying cycle, especially with regards to sperm preservation may allow for the development of industry-based protocols for semen storage and cryopreservation that mimic the sperm preservation capabilities of SST and improve fertility.

Advances in storage of poultry semen

Semen cryopreservation is a key biotechnological strategy used to preserve and protect genetic resources, which are subject to increasingly serious reductions in some species, and to protect animal biodiversity. Assisted reproductive techniques, however, are still not utilized to the same extent in avian species to the extent that occurs in mammals. The reasons for this situation are described in this review. The content of this paper is focused on current poultry preservation systems, published since 2010, and new strategies that are very promising for preserving avian genetic resources. Two major types of storage technologies which are utilized for avian sperm preservation, liquid storage and cryopreservation, are emphasized. The issues on which there is a focus includes supplementation of avian extenders with various compounds prior to the preservation period, use of cryoprotectants and fertility results when there were in vitro sperm evaluations. Results from recent studies indicate there are opportunities to improve the quality of bird semen after preservation. It is obvious that cryo-diluent composition may be the most important factor for development of efficacious cryopreservation methods for avian semen.

Withania somnifera Chemical Constituents' In Vitro Antioxidant Potential and Their Response on Spermatozoa Parameters.

Background: Phytomedicine is becoming more acceptable as an alternative medicinal approach in the modern era.Objectives: The current study examined the antioxidant capacity and in vitro response of phytochemical constituents of Withania somnifera (Ashwagandha) on standard parameters of healthy volunteer semen.Methods: The phytochemicals and their pharmacological response in a hydroethanolic (30:70 v/v) extract of W. somnifera roots were determined using standard protocols.Results: The constituents included flavonoids, phenolic acids, alkaloids, and terpenoids were reported. High-performance liquid chromatography and Fourier-transform infrared spectroscopy determined a diverse array of biologically active chemical constituents in the extract. The extract of W. somnifera exhibits substantial antioxidant properties, including total antioxidant capacity, 2,2-diphenyl-1-picrylhydrazyl inhibition, H2O2 scavenging, and Fe3+ reducing potential (P < .05). The analysis of essential natural minerals explored adequate levels determined using atomic absorption spectrophotometer. Cytotoxic studies revealed significant thrombolytic, RBC membrane stabilization, and DNA damage protection activity (P < .05) while remaining non-mutagenic against Salmonella typhi TA98 and TA100. The best protective response of W. somnifera extract on human semen parameters (n = 30), such as total motility, progressive motility, and viability, demonstrated a significant (P < .05) improvement, particularly at the dose of 25 μg/mL and 50 μg/mL.Conclusion: The study concludes that W. somnifera possesses favorable in vitro characteristics that could aid in the preservation of sperm during intrauterine insemination and in vitro fertilization.

Sperm Characteristics in Microminipigs.

The microminipig is a relatively new type of mini pig; microminipigs weigh about 10 kg at 6 months of age and are expected to be of use in drug discovery research and safety tests. Herein, we analyzed the characteristics of ejaculated sperm from microminipigs. Sperm parameters such as microstructure and sensitivity to cold shock were investigated using optical or scanning electron microscopy. Ejaculate volumes and total numbers of sperm were lower than in standard pig strains, but were proportional to body weight. Ejaculation time, pH of the ejaculate, sperm motility and morphology, and sensitivity to cold shock were similar to those of standard pig strains. Herein, we provide the first characterization of the ejaculates of microminipigs and demonstrate that this type of pig will be useful not only in medical research, but also in investigations into sperm preservation in different pig breeds.

Reconstruction of spermatogonial niche for male fertility preservation

Infertility is one of the late side effects of cancer treatment. Expansion of anti-cancer treatment allow patients to have more life time, however infertility is becoming a matter damaging QOL during the young cancer survivors. The passive strategy such as avoiding the gonad-toxic drug or decreasing the total volume of them and shielding the gonads against cancer therapy has been conducted. To preserve the fertility of young female, ovary tissue cryopreservation is becoming a standard over the world after the success of offspring from cryopreserved ovary tissue autograft was reported. Sperm preservation method is established for the male fertility preservation method, however this is only applicable for sexually matured male patients. For the sake of preserving fertility of sexually immature male patients, many trials using cryopreserved testis tissues or testicular cells have been undergone. Recently, in vitro gametogenesis from stem cell of the human and the mouse to primordial germ cell like cell has been achieved. Here the previous challenges and the latest reports for obtaining functional sperm from immature testis and the reconstruction of spermatogonial niche as a potential approach for preserving fertility procedure are described.

Drug release and sperm motility protection studies of vitamin E encapsulated in liposome, cyclodextrin or polyethylene glycol

Nanoencapsulation may be a good way to preserve hydrophobic bioactive components from unfavorable conditions while also increasing their solubility and bioavailability. Vitamin E has major hurdles because of these chemicals' limited solubility and bioavailability. In this study, the drug release and sperm motility effects of several nanoencapsulation delivery strategies for alphatocopherol, such as inclusion complexes through cyclodextrins, liposomes, and PEG, are investigated. The alpha tocopherol release test was performed on the dialysis bags, and the motility parameters were analyzed using Casa. Because of the rigidity of the liposomal barrier, which was enhanced by adding cholesterol, liposomes did not release the vitamin E immediately, but instead kept it for 24 h. After that, within 48 h, all of the vitamin E molecules had been released. Due to both agitation and fence hydration, release was most likely by Fickian diffusion, which involved the formation of mini-ducts. Furthermore, compared to all other therapies, semen motility improved significantly when vitamin E liposome formulations were employed. Liposomes developed in this study modified the release of vitamin E, making them a promising option for sperm preservation during cryopreservation.

Effect of Sperm Cryopreservation on miRNA Expression and Early Embryonic Development.

Although sperm preservation is a common means of personal fertility preservation, its effects on embryonic development potential need further investigation. The purpose of this study was to identify key microRNA (miRNA) in cryopreserved sperm and determine the changes of these miRNAs and their target genes during embryonic development using cryopreserved sperm. Moreover, the embryonic development potential of cryopreserved sperm was estimated in assisted reproductive technology (ART), where key miRNAs and target genes were validated in sperm and subsequent embryos. Clinical data of embryonic development from cryopreserved sperm indicated a significant decrease in fertilization rate in both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cases, as well as a reduction in blastocyst formation rate in ICSI cases. Meanwhile there was a significant increase in blocked embryo ratio of Day1, Day2, and Day3.5 embryos when frozen-thawed mouse sperm was used, compared with fresh mouse sperm, suggesting a potential negative effect of sperm cryopreservation on embryonic development. From frozen-thawed and fresh sperm in humans and mice, respectively, 21 and 95 differentially expressed miRNAs (DEmiRs) were detected. miR-148b-3p were downregulated in both human and mouse frozen-thawed sperm and were also decreased in embryos after fertilization using cryopreserved sperm. Target genes of miR-148b-3p, Pten, was identified in mouse embryos using quantitative real-time PCR (qRT-PCR) and Western blot (WB). In addition, common characters of cryopreservation of mouse oocytes compared with sperm were also detected; downregulation of miR-148b-3p was also confirmed in cryopreserved oocytes. In summary, our study suggested that cryopreservation of sperm could change the expression of miRNAs, especially the miR-148b-3p across humans and mice, and may further affect fertilization and embryo development by increasing the expression of Pten. Moreover, downregulation of miR-148b-3p induced by cryopreservation was conserved in mouse gametes.

Oleuropein Effectively Improves the Quality of Pig Sperm at 17o C

Artificial Insemination (AI) has been widely used in pig reproduction because of its low price and high efficiency. Cryopreservation of sperm not only leads to a sharp decline in sperm motility, but had high price in pig reproduction. Therefore, most of farms stored pig sperm at 17°C. During the preservation of sperm, the accumulation of Reactive Oxygen (ROS) is the main reason for the decline of sperm motility. The aim of our study was to reduce the accumulation of ROS and improve sperm quality by adding oleuropein (OLE, CAS No. 32619- 42-4) during the storage. The sperm samples were collected and diluted with different concentrations of oleuropein (0, 12, 24, 36, 48, 60 μmol/L). Sperm motility, plasma membrane integrity, acrosome integrity, sperm Total Antioxidant Capacity (T-AOC), Malondialdehyde (MDA) content, Catalase (CAT) activity and Reactive Oxygen Species (ROS) content of boar sperm were measured. The results suggested that adding 36μM of OLE samples significantly improved sperm quality during preservation at 17o C.

Efektivitas Low Density Lipoprotein (LDL) dari Kuning Telur Ayam terhadap Kualitas Semen Cair Domba

Low-Density Lipoprotein (LDL) extracted from egg yolk (LDL) has recently known can eliminate the adverse effect associated in the use of fresh egg yolk. The role of LDL in liquid preservation at 4°C of ram sperm has not been explored. This research evaluates the effects of substituting egg yolk with LDL in ram sperm preservation at 4 °C on 5 days. The objective of this research was to assess the effects of substituting egg yolk with LDL for use as an extender in sperm preservation at 4°C, as well as on spermatozoa motility, viability, morphology, plasma membrane, and acrosome integrity, for 5 days. The semen was subsequently divided into five and diluted with Tris–fresh egg yolk (K), Tris–LDL5% (LDL5), Tris–LDL10% (LDL10), Tris–LDL15% (LDL15), and Tris–LDL20% (LDL20). The result showed a significant difference between LDL to fresh egg yolk for ram sperm quality (P&lt;0.05). The effectiveness of LDL on sperm quality decreased following by its concentration. Even though up to 20% concentration of LDL, it can not preserve the quality of diluted semen for motility, viability, and plasm membrane integrity.

Histological study on the influence of puberty suppression and hormonal treatment on developing germ cells in transgender women.

STUDY QUESTIONCan transgender women cryopreserve germ cells obtained from their orchiectomy specimen for fertility preservation, after having used puberty suppression and/or hormonal treatment?SUMMARY ANSWERIn the vast majority of transgender women, there were still immature germ cells present in the orchiectomy specimen, and in 4.7% of transgender women—who all initiated medical treatment in Tanner stage 4 or higher—mature spermatozoa were found, which would enable cryopreservation of spermatozoa or testicular tissue after having used puberty suppression and/or hormonal treatment.WHAT IS KNOWN ALREADYGender affirming treatment (i.e. puberty suppression, hormonal treatment, and subsequent orchiectomy) impairs reproductive function in transgender women. Although semen cryopreservation is generally offered during the transition process, this option is not feasible for all transgender women (e.g. due to incomplete spermatogenesis when initiating treatment in early puberty, in case of inability to masturbate, or when temporary cessation of hormonal treatment is too disruptive). Harvesting mature spermatozoa, or testicular tissue harboring immature germ cells, from orchiectomy specimens obtained during genital gender-affirming surgery (gGAS) might give this group a chance of having biological children later in life. Previous studies on spermatogenesis in orchiectomy specimens showed conflicting results, ranging from complete absence of germ cells to full spermatogenesis, and did not involve transgender women who initiated medical treatment in early- or late puberty.STUDY DESIGN, SIZE, DURATIONHistological and immunohistochemical analyses were performed on orchiectomy specimens from 214 transgender women who underwent gGAS between 2006 and 2018. Six subgroups were identified, depending on pubertal stage at initiation of medical treatment (Tanner stage 2-3, Tanner stage 4-5, adult), and whether hormonal treatment was continued or temporarily stopped prior to gGAS in each of these groups.PARTICIPANTS/MATERIALS, SETTING, METHODSAll transgender women used a combination of estrogens and testosterone suppressing therapy. Orchiectomy specimen sections were stained with Mayer’s hematoxylin and eosin and histologically analyzed to assess the Johnsen score and the ratio of most advanced germ cell types in at least 50 seminiferous tubular cross-sections. Subsequently, immunohistochemistry was used to validate these findings using spermatogonia, spermatocytes or spermatids markers (MAGE-A3/A4, γH2AX, Acrosin, respectively). Possibilities for fertility preservation were defined as: preservation of spermatozoa, preservation of spermatogonial stem cells or no possibilities (in case no germ cells were found). Outcomes were compared between subgroups and logistic regression analyses were used to assess the association between the duration of hormonal treatment and the possibilities for fertility preservation.MAIN RESULTS AND THE ROLE OF CHANCEMature spermatozoa were encountered in 4.7% of orchiectomy specimens, all from transgender women who had initiated medical treatment in Tanner stage 4 or higher. In 88.3% of the study sample orchiectomy specimens only contained immature germ cells (round spermatids, spermatocytes or spermatogonia, as most advanced germ cell type). In 7.0%, a complete absence of germ cells was observed, all these samples were from transgender women who had initiated medical treatment in adulthood. Cessation of hormonal treatment prior to gGAS did not affect the presence of germ cells or their maturation stage, nor was there an effect of the duration of hormonal treatment prior to gGAS.LIMITATIONS, REASONS FOR CAUTIONSince data on serum hormone levels on the day of gGAS were not available, we were unable to verify if the transgender women who were asked to temporarily stop hormonal treatment 4 weeks prior to surgery actually did so, and if people with full spermatogenesis were compliant to treatment.WIDER IMPLICATIONS OF THE FINDINGSThere may still be options for fertility preservation in orchiectomy specimens obtained during gGAS since a small percentage of transgender women had full spermatogenesis, which could enable cryopreservation of mature spermatozoa via a testicular sperm extraction procedure. Furthermore, the vast majority still had immature germ cells, which could enable cryopreservation of testicular tissue harboring spermatogonial stem cells. If maturation techniques like in vitro spermatogenesis become available in the future, harvesting germ cells from orchiectomy specimens might be a promising option for those who are otherwise unable to have biological children.STUDY FUNDING/COMPETING INTERESTNone.TRIAL REGISTRATION NUMBERN/A.

Cryopreservation of sperm of Labeobarbus brevicephalus (Pisces: Cyprinidae) from Lake Tana (Ethiopia)

The cryopreservation of the Lake Tana Labeobarbus semen had never been studied before. A combination of three extenders and two cryoprotectants (DMSO and glycerol) were used in Lake Tana Labeobarbus brevicephalus sperm cryopreservation. Among the above described cryo-diluent DMSO 10% had the best performance with all three extenders. Sperm of the L. brevicephalus maintained a higher fertilization rate in the three extenders at a dilution ratio of 1:3 (≥ 85%), exceptionally in Extender 2 DMSO 15% sperm displayed lower value 55.9 ± 0.3%. When pre-frozen cryoprotectants milt diluted at a ratio of 1:9, it provided higher fertilization rates in Extender 1 10% DMSO, Extender 3 glycerol 10% and DMSO 15% (92–97%). Highest equilibration (87.3 ± 1.5%) and post-thaw (83.0 ± 1%) motility were obtained from the diluent, Extender 3 plus DMSO 10%, which proved its suitability for preservation of sperm of the L. brevicephalus. The developed protocol suggests formulating an extending medium comprising of Extender 3 with 10% DMSO, 15% DMSO or Extender 1 with 10% DMSO and the dilution ratio of the cryoprotectant to milt should be 1:3. Two different single-step cooling rates (4 and 10 °C/min) were used. Additional studies should evaluate the viability, survival, and development of larvae produced from cryopreserved sperm. Besides, for sustainable use of sperm suitable oocyte sperm ratios and the feasibility of cryogenic gene banking should be investigated.

“Fine feathers make fine birds” – The mammalian sperm plasma membrane lipid composition and effects on assisted reproduction

The sperm plasma membrane is important in modulating many sperm functions. The sperm membrane is composed of a complex mixture of lipids including phospholipids, glycolipids and sterols. There are differences of sperm membrane composition among mammalian species with two groups differing in the most abundant polyunsaturated fatty acid (PUFA), either docosahexaenoic (ω-3 PUFA) or docosapentaenoic acid (ω-6 PUFA). During testicular and epididymal maturation, composition of the sperm plasma membrane evolves with spermatozoa gaining the capacity for fertilization. The importance of fatty acid metabolism for complete spermatogenesis has been elucidated using gene knockout mice. During epididymal transit cholesterol content decreases and PUFA content increases, conferring more fluidity properties to the sperm membrane. The relatively lesser content of antioxidant enzymes and the relatively larger content of PUFA make the spermatozoa particularly susceptible to lipid peroxidation during sperm preservation. In numerous studies, there was adding of PUFA and antioxidants to the diet of animals or to semen extenders with the aim to improve sperm membrane integrity. This review highlights the current knowledge on the sperm membrane composition and effects on sperm function in mammalian domestic animals.

Male-Specific Late Effects in Adult Hematopoietic Cell Transplantation Recipients: A Systematic Review from the Late Effects and Quality of Life Working Committee of the Center for International Blood and Marrow Transplant Research and Transplant Complications Working Party of the European Society of Blood and Marrow Transplantation

Male-specific late effects after hematopoietic cell transplantation (HCT) include genital chronic graft-versus-host disease (GVHD), hypogonadism, sexual dysfunction, infertility, and subsequent malignancies, such as prostate, penile, and testicular cancer. These effects may be closely intertwined and cause prolonged morbidity and decreased quality of life after HCT. Here we provide a systematic review of male-specific late effects in a collaboration among transplantation physicians, endocrinologists, urologists, dermatologists, and sexual health professionals through the Late Effects and Quality of Life Working Committee of the Center for International Blood and Marrow Transplant Research and the Transplant Complications Working Party of the European Society of Blood and Marrow Transplantation. We used a systematic review methodology to summarize incidence, risk factors, screening, prevention, and treatment of these complications and provide consensus evidence-based recommendations for clinical practice and future research. Most of the evidence regarding male GVHD is still based on limited data, precluding strong therapeutic recommendations. Therefore, we recommend systematic screening for male genital GVHD regularly and reporting of cases to large registries to allow for a better understanding. Future research also should address treatment, given the little published evidence currently available. Male-specific endocrine consequences of HCT include hypogonadism, which also may affect bone health. Given the scanty evidence, current recommendations for hormone substitution and/or bone health treatment are based on similar principles as for the general population. Following HCT, sexual health decreases, and this topic should be addressed at regular intervals. Future studies should focus on interventional strategies to address sexual dysfunction. Infertility remains prevalent in patients having undergone myeloablative conditioning, warranting the offer of sperm preservation for all HCT candidates. Most studies on fertility rely on descriptive registry analysis and surveys, underscoring the importance of reporting post-HCT conception data to large registries. Although the quality of evidence is low, the development of cancer in male genital organs does not seem more prevalent in HCT recipients compared with the general population; however, subsequent malignancies in general seem to be more prevalent in males than in females, and special attention should be given to skin and oral mucosa. Male-specific late effects, which likely are more underreported than female-specific complications, should be systematically considered during the regular follow-up visits of male survivors who have undergone HCT. Care of patients with male-specific late effects warrants close collaboration between transplantation physicians and specialists from other involved disciplines. Future research should be directed toward better data collection on male-specific late effects and on studies about the interrelationships among these late effects, to allow the development of evidence-based effective management practices.