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Sperm Cryopreservation (sperm + cryopreservation)
Selected AbstractsSperm Cryopreservation in Brown Bear (Ursus arctos): Preliminary AspectsREPRODUCTION IN DOMESTIC ANIMALS, Issue 2008L Anel Contents The development of sperm cryopreservation procedures in brown bear is the basis for establishing a specific genetic resource bank aimed at the preservation of a Cantabric brown bear population, which is seriously threatened. Several issues complicate the development of these cryopreservation procedures: lack of previous specific studies, a high incidence of urospermia and spermagglutination observed in bear ejaculates. Moreover, the availability of individuals for research from these threatened populations is problematic. In the case of the Cantabric brown bear, we have used males from other populations, but of the same species, as surrogates, to carry out a direct extrapolation of the results. Urospermia , Moreover, 70% of the ejaculates are urine contaminated and spermagglutination have a detrimental effect on post-thawing cell quality recovery in this species. Considering the high value of these samples (autochthonous population with few individuals), a pre-selection of the ejaculates is not a viable alternative. Preventive methods reducing the mentioned detrimental effects need to be developed. On the basis of previous data, we can suppose that bear spermatozoa resist freezing injuries well. Nevertheless, because of the scarcity of this information, it is necessary to conduct further research on bear semen freezing under field conditions. Epidydimal spermatozoa can be important for genetic resource banking of threatened populations and thus specific cryobiological protocols need to be assayed. To date, 168 brown bear ejaculates have been frozen by the ITRA-ULE group at the University of León (Spain) in the development of methodologies for the preservation of brown bear sperm. [source] Cryopreservation of fish sperm: applications and perspectivesJOURNAL OF APPLIED ICHTHYOLOGY, Issue 5 2010E. Cabrita Summary Cryopreservation is of interest not only for fish farming but also for the conservation and genetic improvement of resources. This technique has been well established in some freshwater fish species mainly, salmonid, sturgeons and carps, however, only in the last decade research was focused in marine fish species. The benefits of sperm cryopreservation include: (i) synchronization of gamete availability of both sexes, (ii) sperm economy; (iii) simplification of broodstock management, (iv) transport of gametes from different fish farms, and (v) germplasm storage for genetic selection programs or conservation of species. These issues would certainly benefit the aquaculture industry. The tremendous impact that biotechnology is having in aquaculture has been particularly obvious in recent years. Several species are being used as research models not only for aquaculture development applications but also for medical research. Sperm cryopreservation can give an important contribution in the germ storage of all transgenic lines. However, in all applications in fish sperm, cryopreservation needs to overcome a lack in standardization of methodologies and procedures, a correct assay of seminal quality and the development of tools to characterize cryoinjury. Many efforts have recently been made in the study of DNA using different approaches such as the comet assay (single cell gel electrophoresis), TUNEL (terminal deoxynucleotidyl transferase-nick-end-labelling), SCSA (sperm chromatin structure assay) and the analysis of specific DNA sequences using RT-PCR, since DNA damage may impair fertility or embryo development. Cryopreservation of gametes would certainly benefit from a higher concern on male improvement, basically through nutrition or selection of resistant stocks (e.g. stress resistant individuals or highly adapted to captivity) producing gametes of higher quality. There is a huge window of opportunities for improve the resistance of cells to cryopreservation through diet supplementation of certain compounds such as amino acids (taurine and hypotaurine), vitamins (Vit. E and C) and lipids or through a direct supplementation of the extender media. An equilibrium of those compounds will improve spermatozoa and seminal plasma composition protecting cells against oxidative stress (lipid peroxidation, protein oxidation, DNA fragmentation, enzyme protection) that is gaining each day more importance in cryodamage research. [source] Penile vibratory stimulation and electroejaculation in the treatment of ejaculatory dysfunction,INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 6 2002JENS SŘNKSEN Summary The purpose of this review is to present the current understanding of penile vibratory stimulation (PVS) and electroejaculation (EEJ) procedures and its clinical use in men with ejaculatory dysfunction. Unfortunately, the record of treating such individuals has been quite poor, but within recent years development and refinement of PVS and EEJ in men with spinal cord injury (SCI) has significantly enhanced the prospects for treatment of ejaculatory dysfunction. The majority of spinal cord injured men are not able to produce antegrade ejaculation by masturbation or sexual stimulation. However, approximately 80% of all spinal cord injured men with an intact ejaculatory reflex arc (above T10) can obtain antegrade ejaculation with PVS. Electroejaculation may be successful in obtaining ejaculate from men with all types of SCI, including men who do not have major components of the ejaculatory reflex arc. Because vibratory stimulation is very simple in use, non-invasive, it does not require anaesthesia and is preferred by the patients when compared with EEJ, PVS is recommended to be the first choice of treatment in spinal cord injured men. Furthermore, EEJ has been successfully used to induce ejaculation in men with multiple sclerosis and diabetic neuropathy. Any other conditions which affect the ejaculatory mechanism of the central and/or peripheral nervous system including surgical nerve injury may be treated successfully with EEJ. Finally, for sperm retrieval and sperm cryopreservation before intensive anticancer therapy in pubertal boys, PVS and EEJ have been successfully performed in patients who failed to obtain ejaculation by masturbation. Nearly all data concerning semen characteristics in men with ejaculatory dysfuntion originate from spinal cord injured men. Semen analyses demonstrate low sperm motility rates in the majority of spinal cord injured men. The data give evidence of a decline in spermatogenesis and motility of ejaculated spermatozoa shortly after (few weeks) an acute SCI. Furthermore, it is suggested that some factors in the seminal plasma and/or disordered storage of spermatozoa in the seminal vesicles are mainly responsible for the impaired semen profiles in men with chronic SCI. Home insemination with semen obtained by penile vibratory and introduced intravaginally in order to achieve successful pregnancies may be an option for some spinal cord injured men and their partners. The majority of men will further enhance their fertility potential when using either penile vibratory or EEJ combined with assisted reproduction techniques such as intrauterine insemination or in-vitro fertilization with or without intracytoplasmic sperm injection. [source] Microsurgical vasoepididymostomy with sperm cryopreservation for future assisted reproductionINTERNATIONAL JOURNAL OF UROLOGY, Issue 12 2000Hatsuki Hibi Abstract Background Although obstructive azoospermia is treatable with microscopic seminal reconstruction, the number of patients who choose to undergo vasoepididymostomy is limited because of recent advances in assisted reproductive technology (ART). We attempted to define the outcome of surgical reconstruction in patients with suspected epididymal obstruction and no previous history of vasectomy. Methods We described 40 eligible end-to-side vasoepididymostomy procedures performed on 24 azoospermic patients who had either bilateral or unilateral epididymal obstruction. Results The overall patency rate following surgery was 54% (13/24) and for four patients (17%), natural intercourse resulted in pregnancy. Two pregnancies were initiated with intracytoplasmic sperm injections using frozen sperm collected during vasoepididymostomy. Conclusions In the era of modern ART, microsurgical vasoepididymostomy with cryopreservation of sperm collected during the operation is recommended for patients with epididymal obstructions. [source] Cryopreservation of fish sperm: applications and perspectivesJOURNAL OF APPLIED ICHTHYOLOGY, Issue 5 2010E. Cabrita Summary Cryopreservation is of interest not only for fish farming but also for the conservation and genetic improvement of resources. This technique has been well established in some freshwater fish species mainly, salmonid, sturgeons and carps, however, only in the last decade research was focused in marine fish species. The benefits of sperm cryopreservation include: (i) synchronization of gamete availability of both sexes, (ii) sperm economy; (iii) simplification of broodstock management, (iv) transport of gametes from different fish farms, and (v) germplasm storage for genetic selection programs or conservation of species. These issues would certainly benefit the aquaculture industry. The tremendous impact that biotechnology is having in aquaculture has been particularly obvious in recent years. Several species are being used as research models not only for aquaculture development applications but also for medical research. Sperm cryopreservation can give an important contribution in the germ storage of all transgenic lines. However, in all applications in fish sperm, cryopreservation needs to overcome a lack in standardization of methodologies and procedures, a correct assay of seminal quality and the development of tools to characterize cryoinjury. Many efforts have recently been made in the study of DNA using different approaches such as the comet assay (single cell gel electrophoresis), TUNEL (terminal deoxynucleotidyl transferase-nick-end-labelling), SCSA (sperm chromatin structure assay) and the analysis of specific DNA sequences using RT-PCR, since DNA damage may impair fertility or embryo development. Cryopreservation of gametes would certainly benefit from a higher concern on male improvement, basically through nutrition or selection of resistant stocks (e.g. stress resistant individuals or highly adapted to captivity) producing gametes of higher quality. There is a huge window of opportunities for improve the resistance of cells to cryopreservation through diet supplementation of certain compounds such as amino acids (taurine and hypotaurine), vitamins (Vit. E and C) and lipids or through a direct supplementation of the extender media. An equilibrium of those compounds will improve spermatozoa and seminal plasma composition protecting cells against oxidative stress (lipid peroxidation, protein oxidation, DNA fragmentation, enzyme protection) that is gaining each day more importance in cryodamage research. [source] Physico-biochemical parameters and protein profiles of sperm from beluga Huso husoJOURNAL OF APPLIED ICHTHYOLOGY, Issue 5 2010P. Li Summary Basic physico-biochemical parameters and protein profiles of sperm from beluga (Huso huso), have been evaluated. The results show a stable spermatozoan velocity (ranging from 136.23 ± 5.63 to 105.78 ± 5.27 ,ms,1) and motility during 2 min post activation, as well as a long duration of the overall spermatozoa motility period (up to 5 min). Mean values were determined for seminal plasma protein concentration (0.29 ± 0.16 mg ml,1), spermatozoa concentration (0.28 ± 0.27 × 109 spz ml,1), seminal plasma osmolality (51.33 ± 4.91 mOsmol kg,1), the pH (8.49 ± 0.01) and Na+ (18.97 ± 3.65 mm), K+ (2.83 ± 1.36 mm), Ca2+ (0.19 ± 0.06 mm), Mg2+ (0.49 ± 0.23 mm) and Cl, (6.33 ± 0.58 mm) concentrations. Moreover, in seminal plasma, five protein bands with molecular weights (MW) of 71, 49, 46, 34, 29 kDa were identified. In spermatozoa, about 100 spots with molecular weights varying from 26.5 to 107 kDa and iso-electric points ranging from 5 to 9.5 were found. The observed physiological and biochemical properties, together with protein patterns, should be considered for the development of methods for controlled reproduction and sperm cryopreservation for the highly endangered beluga sturgeon. [source] The current status of sperm cryopreservation of the endangered Probarbus jullieni (Sauvage) in MalaysiaJOURNAL OF APPLIED ICHTHYOLOGY, Issue 5 2010P. C. Chew Summary The objective of this study was to develop a cryopreservation method in Probarbus jullieni sperm, an endangered riverine fish species in Southeast Asia, including the optimization of an extender solution (14 extender formulations were tested) and selecting a cryoprotectant (five types of agents and methanol were used at concentrations (v/v) of 5, 7.5, 9, 10, 12, 15 and 20%). The semen to diluent ratios tested were as follow: 1 : 1, 1 : 2, 1 : 3, 1 : 4, 1 : 5, 1 : 7, 1 : 9, 1 : 14, 1 : 19, 1 : 24 and 1 : 49. Vapour exposure duration was set at 5, 10, 15 and 20 min while the distance between sample and liquid nitrogen (LN2) during the vapour exposure was designed at 3, 3.5, 4, 5 and 6 cm. Further, the time frame for thawing was set at 6, 7, 8, 10, 20 and 30 s. The optimum protocol was by using CF-HBSS (pH 7.5, osmolality 285 ± 10 mOsmol kg,1) in combination with methanol at 9% (v/v); sperm to diluents ratio between 1 : 3 to 1 : 5; vapour exposure for 5 min or 10 min, with samples placed at 3.5 cm or 4 cm above LN2 and thawing at 40°C for 7 s. The mean of pre-frozen and post-thaw sperm motility was 80.1 ± 13.6% (n = 43) and 49.6 ± 16.4% (n = 43) respectively. The reproductive characteristics of P. jullieni during its spawning season were addressed in present work. Cryopreserved sperm was found to have lower fertilization ability (4.2 ± 2.5%, n = 1050) and hatching rate (1.6 ± 1.2%, n = 1050) compared with fresh sperm (fertilization 77.7 ± 6.2%, n = 1050; hatching 64.7 ± 7.7%, n = 1050). The resulted problems and constraints encountered in the process of sperm cryopreservation of the species studied were also reported in this paper. [source] Improved Cryopreservation of Sperm of Paddlefish (Polyodon spathula)JOURNAL OF THE WORLD AQUACULTURE SOCIETY, Issue 4 2006ÁKos Horváth Experiments were performed to improve protocols for sperm cryopreservation of paddlefish (Polyodon spathula), a species for which there has been limited study. The first experiment was conducted to investigate the effects of two extenders (modified Tsvetkova's extender: mT and modified Hanks' balanced salt solution: mHBSS) in combination with methanol (MeOH) and dimethyl sulfoxide in two concentrations (5 and 10%) on the postthaw motility and fertilization rates of cryopreserved sperm. The highest postthaw motility (85 ± 5%) was observed when sperm were frozen using mT extender with 10% MeOH as cryoprotectant. Extenders (P = 0.0018) and cryoprotectants (P = 0.0040) each had a significant effect on the postthaw motility of paddlefish sperm. The highest fertilization (80 ± 3%) was found when eggs were fertilized with sperm frozen with mT extender in combination with 10% MeOH. However, there was no significant difference among fertilization rates when MeOH was used as a cryoprotectant in either concentration or in combination with either mT or mHBSS extenders. In the second experiment, 4000 eggs were fertilized with the pooled contents of five straws of thawed sperm (total volume of 1.25 mL) using mT extender in combination with 5% MeOH, and hatch rates as high as 79 ± 5% were observed. A third experiment was also conducted to clarify the role of MeOH concentration; however, no significant difference was found among fertilization and hatch rates when either 5 or 10% MeOH was used as a cryoprotectant. These results suggest that MeOH is a safe and reliable cryoprotectant for freezing of paddlefish sperm and obtaining viable postthaw sperm for consistent fertilization and hatch rates. Further, this experimental protocol is relatively simple and applicable for commercial hatchery production of paddlefish. [source] Comparative studies with six extenders for sperm cryopreservation in the cynomolgus monkey (Macaca fascicularis) and rhesus monkey (Macaca mulatta)AMERICAN JOURNAL OF PRIMATOLOGY, Issue 1 2006Yahui Li Abstract Ejaculated spermatozoa from cynomolgus monkeys and rhesus monkeys were frozen in straws with six different extenders (TTE, DM, mDM, LG-DM, G-DM, and TCG) containing glycerol. Sperm motility and head membrane and acrosomal integrity were evaluated after freezing and thawing, and the cryoprotective effects were compared among the extenders and the two species studied. The results showed that sperm motility and motility recovery with the six extenders were comparable for the cynomolgus and rhesus monkeys. There was no significant difference in sperm motility and head membrane integrity among the six extenders in either the cynomolgus or rhesus monkeys (P>0.05). However, a slightly but statistically lower percentage of acrosomal integrity was found with TCG in both species compared to the other extenders (P<0.05). These findings demonstrate that TTE, DM, mDM, LG-DM, G-DM, and TCG are equally suitable extenders for the cryopreservation of spermatozoa from cynomolgus and rhesus monkeys. Am. J. Primatol. 68:39,49, © 2006 Wiley-Liss, Inc. [source] Effect of amino acids on cryopreservation of cynomolgus monkey (macaca fascicularis) spermAMERICAN JOURNAL OF PRIMATOLOGY, Issue 4 2003Yahui Li Abstract The effects of three amino acids (proline, glutamine, and glycine) added to the freezing medium Tes-Tris-egg yolk (TTE) for cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa were studied. This is the first report on the effects of amino acids on nonhuman primate sperm cryopreservation. The addition of 5mM proline, 10mM glutamine, and 10 or 20mM glycine each significantly improved post-thaw sperm motility and membrane and acrosome integrity compared with the control (TTE alone). However, a significant decrease in motility and membrane/acrosome integrity was observed when amino acid concentrations increased to 60mM for proline and glutamine, and 80mM for glycine. The results suggest that adding a limited amount of amino acids to the freezing media is beneficial for freezing cynomolgus monkey sperm. Am. J. Primatol. 59:159,165, 2003. © 2003 Wiley-Liss, Inc. [source] Bacterial risk and sperm cryopreservationANDROLOGIA, Issue 5 2004R. Levy Summary. Prior to sperm cryopreservation, French guidelines only recommend viral screening for serological status towards human immunodeficiency virus, hepatitis B and C viruses and Treponema palidum. The probability of semen infection by other bacterial pathogens is not taken into consideration by the current recommendations. The objective of the present study was to evaluate this risk and a strategy to reduce it prospectively. Ninety-six patients consulting for sperm cryopreservation underwent a semen culture simultaneously to cryopreservation. The patients were classified into three groups following semen culture results: negative culture (group 1, 77/96, 80.2%), positive culture with saprophytic agents (group 2, 9/96, 9.4%) and positive culture with pathogen agents (group 3, 10/96, 10.4%). For six patients of the latter group showing a genital infection with Ureaplasma urealyticum, a discontinuous gradient selection performed on the cryopreserved sample was efficient to discard bacteria. These data emphasize the usefulness to cultivate semen simultaneously to cryopreservation and demonstrate the ability to remove some microbial agents from semen before its use in assisted reproductive techniques. [source] Effect of storage time and cryoprotectant concentrations on the fertilization rate and hatching rate of cryopreserved sperm in red seabream (Pagrus major Temminck & Schlegel, 1843)AQUACULTURE RESEARCH, Issue 9 2010Qing Hua Liu Abstract This study examined the effects of storage time and cryoprotectant concentrations on the post-thaw sperm of red seabream, Pagrus major. Sperm treated with 12%, 15%, 18% and 21% DMSO were cryopreserved for 10, 30, 60 and 360 days, and fertilization and hatching rates were analysed. For all groups, there were no differences in the fertilization rates and hatching rates between sperm cryopreserved for <60 days and fresh sperm (98.8±0.8%, 96.4±1.3%). However, for sperm cryopreserved for 360 days, both fertilization rates (88.6±3.0% to 7.0±1.9%) and hatching rates (79.4±7.2% to 3.3±0.8%) decreased drastically. Furthermore, the cryoprotectant concentrations affected sperm quality significantly (P<0.05). When cryopreserved for 360 days, sperm treated with 15% DMSO obtained the best results compared with other concentrations. We suggest that 15% DMSO may be an effective cryoprotectant for long-term sperm cryopreservation of red seabream. [source] Cryopreservation of sperm in marine fishAQUACULTURE RESEARCH, Issue 3 2000M. Suquet Since the first work of Blaxter in 1953, fish sperm cryopreservation has been attempted on about 30 marine species. The present paper reviews the techniques used and the results published in these species. Particular attention is paid to the handling procedure of sperm before freezing, the problems of semen ageing and semen contamination with urine. The quality of frozen,thawed semen was evaluated using previously standardized biotests, such as a two-step motility activation technique adapted for the different species and fertilization assays using a discriminating insemination technique. Most extenders used in marine fish are saline or sugar solutions. From the investigated cryoprotectants, dimethyl sulphoxide (DMSO) generally leads to the best results. Cooling rates range from 8 °C to 99 °C min,1; the thawing rate is generally high. Compared with freshwater species, a high percentage of spermatozoa survives cryopreservation. Therefore, and because of the simplicity of the techniques, the cryopreservation of marine fish sperm is suited for application in aquaculture. [source] | |||||||||||||