scholarly journals Evaluation of the Efficiency of Two Different Freezing Media and Two Different Protocols to Preserve Human Spermatozoa from Cryoinjury

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Gemma Fabozzi ◽  
Maria Flavia Starita ◽  
Emilia Rega ◽  
Alessandra Alteri ◽  
Antonio Colicchia ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Human Sperm ◽  
Human Spermatozoa ◽  
Sperm Cryopreservation ◽  
Progressive Motility ◽  
Direct Addition ◽  
Sperm Sample ◽  
Sperm Survival ◽  
Better Than

It is universally recognized that cryopreservation impairs sperm quality. In order to improve postthawing sperm survival and motility, media of different composition and different protocols have been proposed. However, no clear evidence is available to understand which are the most efficient protocol and medium for sperm cryopreservation. The present study evaluates the efficiency of two different cryopreservation protocols and two common freezing media (FM) containing different cryoprotectants (CPs), TEST Yolk Buffer (TYB) and Sperm Freeze (SF), to preserve human sperm quality. Our data suggest that TYB is better than SF both in terms of postthaw viability and in terms of progressive motility, while the direct addition of FM to the sperm sample resulted in the most efficient protocol in terms of postthaw viability but not in terms of progressive motility.

scholarly journals Effects of human sperm cryopreservation on apoptotic markers in normozoospermic and non-normozoospermic patients

Zygote ◽  
2018 ◽  
Vol 26 (4) ◽  
pp. 308-313 ◽  
Author(s):  
Seda Karabulut ◽  
Asuman Demiroğlu-Zergeroğlu ◽  
Elif Yılmaz ◽  
Pelin Kutlu ◽  
İlknur Keskin
Keyword(s):  
Dna Fragmentation ◽  
Caspase 3 ◽  
Sperm Quality ◽  
Human Sperm ◽  
Sperm Parameters ◽  
Semen Parameters ◽  
Sperm Cryopreservation ◽  
Negative Effects ◽  
Apoptotic Markers ◽  
Neat Semen

SummaryThe negative effects of cryopreservation on sperm parameters are well documented but little information is known about molecular basis of the process. The aim of the present study was to investigate the possible effects of sperm cryopreservation on main apoptotic signs including DNA fragmentation and caspase-3 activation and to determine if these effects vary according to sperm parameters. Sperm samples of 72 patients were cryopreserved. The patients were sub-grouped as normozoospermic or non-normozoospermic patients according to their semen parameters. DNA fragmentation rates and caspase-3 activation levels were analyzed before and after cryopreservation in both groups. Mean DNA fragmentation rate was increased significantly from 23.98% in neat semen samples to 27.34% after cryopreservation (P = 0.03). DNA fragmentation rates were slightly higher in non-normozoospermic patients compared with the normozoospermic patients in both the neat semen and after cryopreservation (23.25 and 24.71% vs. 26.32 and 28.36%, respectively) although the difference obtained were not statistically significant. An increasing trend for caspase-3 activations (0.093 vs. 0.116) was observed after cryopreservation but the differences were not statistically significant. Caspase-3 activation was found to be slightly higher in non-normozoospermic patients both in the neat semen and after cryopreservation compared with the normozoospermic patients but the differences were not statistically significant. Caspase-3 expression was also shown using immunocytochemistry in both fresh ejaculated sperm and thawed sperm after cryopreservation but at different localizations. The cryopreservation process had detrimental effects on sperm quality but the quality of the sperm samples was not adversely effective for the apoptotic markers including DNA fragmentation and caspase-3 activation patterns. In fact, it was the cryopreservation process itself that adversely effected the above apoptotic markers and apoptosis. It was concluded therefore that sperm cell cryopreservation triggers apoptosis after thawing and this process adversely affects semen parameters.

Testis developmental related gene 1 (TDRG1) encodes a progressive motility-associated protein in human spermatozoa

2020 ◽  
Author(s):  
Houyang Chen ◽  
Liang Tang ◽  
Qing Hong ◽  
Tingting Pan ◽  
Shiqi Weng ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Conflicts Of Interest ◽  
Human Sperm ◽  
Human Spermatozoa ◽  
Mass Spectrometry Analysis ◽  
Human Testis ◽  
Specific Gene ◽  
Related Gene ◽  
Progressive Motility ◽  
Related Proteins

Abstract STUDY QUESTION Is there an association between the human testis-specific gene, testis developmental related gene 1 (TDRG1) and human sperm motility? SUMMARY ANSWER TDRG1 is associated with asthenozoospermia and involved in regulating human sperm motility. WHAT IS KNOWN ALREADY Many testis-specific proteins potentially regulate spermatogenesis and sperm motility. We have identified a novel human testis-specific gene, TDRG1, which encodes a 100-amino-acid protein localized in the human sperm tail, yet little is known about its role in human spermatozoa. STUDY DESIGN, SIZE, DURATION Sperm samples were obtained from normozoospermic men and asthenozoospermic men who visited the reproductive medical center at Jiangxi Maternal and Child Health Hospital, Nanchang, Jiangxi, China between February 2018 and January 2019. In total, 27 normozoospermic men and 25 asthenozoospermic men were recruited to participate in the study. PARTICIPANTS/MATERIALS, SETTING, METHODS The level of TDRG1 in sperm of normozoospermic and asthenozoospermic men was examined by immunoblotting and immunofluorescence assays. Progressive motility was examined by computer-aided sperm analysis. The correlation between the TDRG1 protein level and progressive motility was analyzed by linear regression. TDRG1 was imported into the sperm of normozoospermic and asthenozoospermic men using a cell-penetrating peptide (CPP)-fused TDRG1 recombinant protein (CPP-TDRG1), and the progressive motility was examined. Also, the altered proteins associated with TDRG1 in asthenozoospermic sperm were detected using label-free quantification method-based quantitative proteomic technology. TDRG1-interacting proteins were identified by co-immunoprecipitation coupled with tandem mass spectrometry analysis. MAIN RESULTS AND THE ROLE OF CHANCE The mean level of TDRG1 was significantly decreased in sperm of asthenozoospermic men compared with normozoospermic men (P < 0.05) and was positively correlated with percentage of progressively motile sperm (r2 = 0.75, P = 0.0001). The introduction of TDRG1 into human sperm, using CPP, significantly increased progressive motility (P < 0.05) and improved the progressive motility of sperm from asthenozoospermic men to the normal level. TDRG1 forms a protein complex with sperm-motility related proteins in human sperm and its downregulation was associated with a decrease in other motility-related proteins. LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION The sample size was limited and larger cohorts are needed for verifying the positive effect of CPP-TDRG1 on human sperm motility. Furthermore, the caution should be paid that a comprehensive safety examination would be performed to evaluate whether CPP-TDRG1 is a possible treatment approach for asthenozoospermia. WIDER IMPLICATIONS OF THE FINDINGS Our results provide new insights into the mechanisms of sperm motility which may contribute to the diagnosis and treatment for asthenozoospermia. STUDY FUNDING/COMPETING INTEREST(S) National Natural Science Foundation of China (81501317 and 81871207 to H.C.; 81771644 to T.L.; 31671204 to X.Z.; 81571432 to Y.T.). The authors have no conflicts of interest to declare.

scholarly journals Expression and Localization of δ-, κ-, and μ-Opioid Receptors in Human Spermatozoa and Implications for Sperm Motility

2006 ◽  
Vol 91 (12) ◽  
pp. 4969-4975 ◽  
Author(s):  
Ekaitz Agirregoitia ◽  
Asier Valdivia ◽  
Arkaitz Carracedo ◽  
Luis Casis ◽  
Javier Gil ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Opioid Receptors ◽  
Receptor Agonist ◽  
Reproductive Tract ◽  
Human Sperm ◽  
Human Spermatozoa ◽  
Endogenous Opioid ◽  
Progressive Motility ◽  
Agonist And Antagonist ◽  
Μ Opioid Receptors

Abstract Context: Endogenous opioid peptides signal through δ-, κ-, and μ-opioid receptors. Some of these peptides such as endorphins and enkephalins are present in the male reproductive tract, but the presence of the corresponding receptors in human sperm cells has not yet been reported. Objective: Our objective was to study the expression and localization of δ-, κ-, and μ-opioid receptors on human spermatozoa and the implication in sperm motility. Methods: The expression of receptors was studied by RT-PCR, Western blot, and immunofluorescence techniques. We evaluated the effects of activation of each opioid receptor by specific agonist and antagonist. Results: Human spermatozoa express δ-, κ-, and μ-opioid receptors. These receptors were located in different parts of the head, in the middle region, and in the tail of the sperm. Progressive motility of spermatozoa, an important parameter to evaluate male fertility, was found to be significantly reduced after incubation with the μ-receptor agonist morphine, whereas this effect was antagonized in the presence of the corresponding antagonist naloxone. The δ-receptor antagonist naltrindole significantly reduced progressive motility immediately after its addition. However, the δ-receptor agonist DPDPE had no significant effect. Finally, neither the κ-receptor agonist U50488 nor its antagonist nor-binaltorphimine significantly affected the progressive motility of human spermatozoa. Conclusion: We report for first time the presence of functional δ-, κ-, and μ-opioid receptors in human sperm membranes. These findings are indicative of a role for the opioid system in the regulation of sperm physiology.

scholarly journals Studies on the basic issues relevant to sperm cryopreservation in humans

2020 ◽  
Vol 14 ◽  
pp. 263349412090937
Author(s):  
Huanhuan Hu ◽  
Xiaowei Shi ◽  
Guojie Ji ◽  
Rui Liu ◽  
Jing Zhang ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Nuclear Dna ◽  
Sperm Concentration ◽  
Human Sperm ◽  
Dna Integrity ◽  
Rapid Freezing ◽  
Sperm Cryopreservation ◽  
Recovery Rates ◽  
Freezing Medium ◽  
Better Than

Rapid freezing and vitrification are becoming popular for sperm freezing in humans; however, basic and critical issues relevant to sperm cryopreservation remain to be resolved. The aims of the present study were to study the effects of osmolality of freezing medium, sperm concentrations, thawing methods, and sugars (sucrose and trehalose) on sperm motility and DNA integrity by rapid freezing using 0.5 ml standard straws loaded with 100 µl sperm each. The results showed that (1) the post-thaw recovery rates of total motility and progressive motility of sperm cryopreserved in freezing medium containing 0.25 M sucrose with 442 mOsm/kg osmolality were significantly higher ( p < 0.05) than that of sperm cryopreserved in freezing medium containing 0.25 M sucrose with 536 mOsm/kg osmolality (36.5 ± 2.8% and 36.9 ± 1.7% versus 30.4 ± 1.9% and 30.3 ± 2.9%, respectively), (2) cryopreservation of both total and progressive motilities was not significantly affected ( p > 0.05) by sperm concentrations in the range from 5 to 20 × 106 sperm/ml, (3) thawing method 37°C for 2 min was better than 42°C for 15 s in terms of post-thaw recovery rates of both total and progressive motilities ( p < 0.05), (4) 0.25 M trehalose was better than 0.25 M sucrose in cryopreserving both total and progressive motilities ( p < 0.05), and (5) sperm nuclear DNA is relatively resistant to the changes of the above factors compared with sperm motility. It was concluded that human sperm can be best cryopreserved by rapid freezing using 0.25 M sucrose or trehalose with osmolality 442 to 457 mOsm/kg at high sperm concentration followed by thawing at 37°C. Trehalose is a stronger cryoprotectant than sucrose for sperm cryopreservation.

Effect of temperature, nutrients, calcium, and cAMP on motility of human spermatozoa

1982 ◽  
Vol 242 (5) ◽  
pp. C304-C311 ◽  
Author(s):  
F. K. Gorus ◽  
R. Finsy ◽  
D. G. Pipeleers
Keyword(s):  
Seminal Fluid ◽  
Gradient Centrifugation ◽  
Cell Movement ◽  
Human Sperm ◽  
Correlation Spectroscopy ◽  
Human Spermatozoa ◽  
Effect Of Temperature ◽  
Dependent Manner ◽  
Calcium Dependent ◽  
Progressive Motility

The motility of human spermatozoa and its regulation were examined on cells isolated from other seminal components and purified into fractions of uniform progressive motility. The percent motile cells and estimates of their translational speed were determined by visual inspection, by stroboscopy, and by photon correlation spectroscopy; microcinematography and gradient centrifugation were occasionally used to clarify discrepancies. The motility of isolated spermatozoa could be maintained for periods up to 24 h at 4 or 37 degrees C; the presence of seminal fluid was not required and even provoked a reversible inhibition at 4 degrees C. Albumin facilitated cell movement between microscopic glass plates but had no effect on progressive motility per se, as evidenced by other techniques. During incubations of up to 2 h, progressive cell motility occurred independently of extracellular glucose and calcium but responded to variations in adenosine 3',5'-cyclic monophosphate and calcium. Dibutyryl cAMP increased forward motility, whereas ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid reversibly immobilized the spermatozoa in a calcium-dependent manner; phosphodiesterase inhibition resulted in increased vibration of sperm heads without any effect on progressive motility. Longer incubation periods required the presence of extracellular nutrients. These experiments further demonstrate that several motility measuring techniques should be used in parallel to distinguish the various components of cell movement, to exclude aspecific effects, and to supplement the shortcomings of each individual technique. Such procedure could clarify the various discrepancies that have been reported so far and should lead to a better understanding of the regulation of human sperm motility.

scholarly journals Effects of Trehalose and Sucrose on Human Sperm Motility, Vitality and Morphology after Cryopreservation

2019 ◽  
Author(s):  
Manaphat Suksai ◽  
Kriengsak Dhanaworavibul
Keyword(s):  
Data Analysis ◽  
Sperm Motility ◽  
Vapor Phase ◽  
Human Sperm ◽  
Sperm Parameters ◽  
Phase Method ◽  
P Value ◽  
Progressive Motility ◽  
R Program ◽  
Better Than

Objective: To compare human sperm motility, vitality and morphology after cryopreservation among sucrose, and different trehalose concentrations.Material and Methods: A total number of 124 normozoospermic semen samples were collected. Each semen sample was divided into 4 portions, and cryopreserved in a human sperm-preserving medium along with cryoprotectants, including; 50 milimolar (mM) trehalose, 100 mM trehalose, 200 mM trehalose and 50 mM sucrose, respectively. All semen samples were frozen by using a vapor phase method. Post-thawed sperm motility, vitality and morphology were assessed. R program was used for data analysis. A p-value of <0.05 was considered statistically significant.Results: Post-thawed semen evaluation indicated that 50 mM trehalose was better than 50 mM sucrose in all sperm parameters, which included progressive motility (p-value=0.037, total motility (p-value<0.001), vitality (p-value<0.001) and morphology (p-value<0.001). The sperm parameters were not significantly different among 100 mM trehalose, 200 mM trehalose and 50 mM sucrose.Conclusion: The use of 50 mM trehalose, as non-permeating cryoprotectant, showed superior post-thaw sperm parameters over sucrose, and other trehalose concentrations.

scholarly journals Melatonin and Caffeine Supplementation Used, Respectively, as Protective and Stimulating Agents in the Cryopreservation of Human Sperm Improves Survival, Viability, and Motility after Thawing compared to Traditional TEST-Yolk Buffer

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Juliana R. Pariz ◽  
Caroline Ranéa ◽  
Rosa A. C. Monteiro ◽  
Donald P. Evenson ◽  
Joël R. Drevet ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Semen Quality ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Human Sperm ◽  
Mitochondrial Activity ◽  
Ros Production ◽  
Stimulant Effect ◽  
Progressive Motility ◽  
Before And After

Cryopreservation processes can damage spermatozoa and impair structural and functional cell characteristics. Plasma, nuclear membranes, and cellular organelles can suffer from the freeze and thaw process. This study evaluates the protective and stimulant effect of melatonin and caffeine supplementation on the functional characteristics of human spermatozoa before and after freezing. Thirty seminal samples from normozoospermic men aged 19–45 years old collected between October 2012 and May 2017 were included. Semen samples were supplemented with either 2 mM melatonin (MEL) prior to cryopreservation, 2 mM caffeine (CAF) in postthaw, or CAF and MEL (CM) in precryopreservation and postthaw, respectively. Kinetics and seminal parameters, mitochondrial activity, DNA fragmentation, and reactive oxygen species (ROS) levels were analyzed before and after cryopreservation. A significant reduction in sperm concentration, total and progressive motility, sperm kinetics, and mitochondrial activity, as well as a significant increase in DNA fragmentation and ROS production in postthaw samples compared to fresh samples, was identified. After administration of a caffeine and/or melatonin supplement, there was a significant increase in progressive motility in the CAF (p=0.005) and CM (p=0.048) groups, as well as mitochondrial activity in the CM group (p<0.05). Cryopreservation has negative effects on overall sperm quality and increases ROS production. A combination of caffeine and melatonin in prefreeze and postthaw sperm samples has proven to be a very effective and simple way to improve semen quality. This will be particularly useful for initial low-quality semen samples, those which suffer the most from the freezing/thawing process.

scholarly journals Oxidative Stress Measurement in Frozen/Thawed Human Sperm: The Protective Role of an In Vitro Treatment with Myo-Inositol

Antioxidants ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 10
Author(s):  
Rosetta Ponchia ◽  
Annunziata Bruno ◽  
Asia Renzi ◽  
Claudia Landi ◽  
Enxhi Shaba ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Structural Changes ◽  
Stress Measurement ◽  
Sperm Quality ◽  
Human Sperm ◽  
2D Electrophoresis ◽  
Sperm Cryopreservation ◽  
Atp Production

Despite its widespread use, sperm cryopreservation induces serious detrimental alterations in sperm function; indeed, it is commonly associated with decreased sperm viability and motility, and DNA fragmentation. Mechanisms of human sperm cryodamage are thought to be multifactorial, but oxidative stress seems to have a prominent role. A huge amount of data supported the cryoprotective effect of different antioxidants able to minimize the detrimental effects of reactive oxygen species (ROS) and improve the quality of spermatozoa. Among others, myo-inositol is one of the most powerful and has been reported to be effective in improving sperm quality and motility when used both in vivo and in vitro. This study aimed to determine the in vitro impact of myo-inositol in ameliorating sperm oxidative status during sperm cryopreservation. In particular, we demonstrated a significant improvement of sperm parameters (vitality and motility) when myo-inositol was added after sperm thawing (p < 0.05). Moreover, we showed that myo-inositol induces a significant increase in oxygen consumption, the main index of oxidative phosphorylation efficiency and ATP production. Finally, by means of 2D-electrophoresis, we demonstrated a significant decrease in the level of carbonyl groups, the main structural changes occurring in conditions of oxidative stress (p < 0.05). In conclusion, the sperm cryopreservation procedure we developed, assuring the reduction of ROS-induced sperm modifications, may improve the in vitro procedure currently used in ART laboratory for sperm cryostorage.

P–088 Sexual functioning is impaired in cancer survivors after cancer therapy

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
E Reiser ◽  
K Vomstein ◽  
S Hofer-Tollinger ◽  
G Pinggera ◽  
E Strassgschwandtner ◽  
...  
Keyword(s):  
Sexual Dysfunction ◽  
Cancer Survivors ◽  
Sexual Functioning ◽  
Reproductive Medicine ◽  
Sperm Quality ◽  
Sperm Count ◽  
Sperm Cryopreservation ◽  
Progressive Motility ◽  
Medical University

Abstract Study question Is impaired sexual functioning correlated to sperm quality in cancer survivors? Summary answer Erectile dysfunction affects 25.0% of cancer survivors, independent of sperm quality. 22.9% of patients show symptoms consistent with a reduced testosterone level. What is known already Gonadotoxic treatment in male cancer patients can end up in reversible or permanent impaired spermatogenesis, testosterone insufficiency, and sexual dysfunction. Study design, size, duration In this prospective single-center study, sexual functioning was assessed in male cancer survivors, who underwent sperm cryopreservation at the Department of Gynecological Endocrinology and Reproductive Medicine, Medical University Innsbruck, Austria from 01/2010 to 12/2018. Sexual functioning was assessed between 03–12/2020 via two questionnaires: Aging Male Score (AMS) and International Index of Erectile Function (IEEF-EF). Participants/materials, setting, methods Thirty-five cancer survivors (testicular cancer: n = 16 [45.7%], hematological malignancies: n = 15 [42.9%], others: n = 4 [n = 11.4%]) filled in two questionnaires (AMS and IEEF-EF) during routine follow-up visit at the Department of Gynecological Endocrinology and Reproductive Medicine, Medical University Innsbruck and the Department of Urology, Medical University Innsbruck, Austria. Moreover, sperm quality was assessed and normozoospermia was defined in accordance with the 2010 WHO criteria (sperm concentration ≥15 million/mL, progressive motility ≥32%, and ≥4% normal morphology). Main results and the role of chance Mean age at sperm cryopreservation and follow-up visit was 25.1±4.2 and 31.9±6.3 years, respectively with a mean follow-up time of 81.4±12.5 months. Rate of erectile dysfunction was low (75.0% no dysfunction, 15.6% low dysfunction, 3.1% low-moderate dysfunction, 3.1% moderate, 3.1% severe dysfunction). Moreover, AMS score indicated no, low, moderate and severe symptoms consistent with a low testosterone level in 77.1%, 8.6%, 2.9%, and 2.9% of patients, respectively. Oligozoospermia was observed in up to 48% of the patients with TM and in only 23% patients with HM. Patients with TM showed significantly reduced sperm count (18.7 × 106/mL [5.3–43.0]) and total sperm count (42.4 × 106/ejaculate [13.3–108.5]) compared to HM (p = 0.03). There was no difference in sexual functioning between patients with HM or TM. Sexual functioning did no correlate with sperm count, progressive motility or morphology. Limitations, reasons for caution Although the study may be limited by its small sample size, it is the first to assess a correlation of sperm quality and sexual dysfunction in cancer survivors. Wider implications of the findings: As every fourth male cancer patient suffers from impaired sexual functioning after gonadotoxic treatment, this important topic should be addressed in clinical and scientific future. Future studies should focus on both, somatic and psychosomatic reasons for sexual dysfunction. Trial registration number none

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