Sperm Cryopreservation

Todorovic, Biljana Popovic; Verheyen, Greta; Vloeberghs, Veerle; Tournaye, Herman

doi:10.1007/978-3-030-47767-7_36

Cited by 3 publications

(2 citation statements)

References 112 publications

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“…The rapid freezing method is based on the direct contact of samples with liquid nitrogen vapor for at least 10 minutes, in this method the sperm are mixed with cryoprotective agent and placed in cryotubes that will be exposed to nitrogen vapors. After the vapor exposure phase; the samples are immersed in liquid nitrogen ( 140 , 141 ). The addition of CPAs to the samples seeks to minimize osmotic damage and prevent intracellular and extracellular ice crystals from forming, however, CPAs have cytotoxic characteristics ( 142 ).…”

Section: Current Strategies To Reestablish Testicular Functionalitymentioning

confidence: 99%

Biomaterials for Testicular Bioengineering: How far have we come and where do we have to go?

et al. 2023

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Traditional therapeutic interventions aim to restore male fertile potential or preserve sperm viability in severe cases, such as semen cryopreservation, testicular tissue, germ cell transplantation and testicular graft. However, these techniques demonstrate several methodological, clinical, and biological limitations, that impact in their results. In this scenario, reproductive medicine has sought biotechnological alternatives applied for infertility treatment, or to improve gamete preservation and thus increase reproductive rates in vitro and in vivo. One of the main approaches employed is the biomimetic testicular tissue reconstruction, which uses tissue-engineering principles and methodologies. This strategy pursues to mimic the testicular microenvironment, simulating physiological conditions. Such approach allows male gametes maintenance in culture or produce viable grafts that can be transplanted and restore reproductive functions. In this context, the application of several biomaterials have been proposed to be used in artificial biological systems. From synthetic polymers to decellularized matrixes, each biomaterial has advantages and disadvantages regarding its application in cell culture and tissue reconstruction. Therefore, the present review aims to list the progress that has been made and the continued challenges facing testicular regenerative medicine and the preservation of male reproductive capacity, based on the development of tissue bioengineering approaches for testicular tissue microenvironment reconstruction.

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Section: Current Strategies To Reestablish Testicular Functionalitymentioning

confidence: 99%

Biomaterials for Testicular Bioengineering: How far have we come and where do we have to go?

et al. 2023

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“…Thus, it is more urgent to search for more affordable methods of assessing the quality of spermatozoa, including the analysis of the morphological specifi cities of their structure, which would be used as an instrument for cryopreservation effi ciency screening. The methods of spermatozoa cryopreservation are known to be suboptimal (Todorovic et al, 2022). It was proven regarding many species of animals that the spermatozoa survival rate after thawing depended on their initial morphofunctional characteristics (Dorado et al, 2010;Casas et al, 2009;Núñez-Martínez et al, 2007;Pavlovych et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Morphological and morphometric characteristics of fresh and cryopreserved spermatozoa of saanen bucks

Bogdaniuk

Petrushko

2023

Agric. sci. pract.

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Background. Cryopreservation of spermatozoa with the best morphofunctional characteristics may allow their use for assisted reproductive technologies in goat breeding throughout the year to effectively increase livestock. Aim. To evaluate the influence of seasons on morphological and morphometric characteristics of fresh and cryopreserved goat sperm. Methods. The ejaculate samples of three sexually mature bucks of the Saanen breed were obtained during the breeding and non-breeding seasons. To assess the morphological characteristics, the smears of sperm were stained and then visualized under a light microscope with a magnification of ×1,000. For morphometric measurements, sperm micrographs were taken and analyzed using the ImageJ software. The cryopreservation of sperm was performed in a medium with 10 % glycerol and 20 % egg yolk. The samples were thawed in a water bath, then the morphological and morphometric characteristics of the cells were evaluated. Results. After the cryopreservation, the number of sperm with abnormal morphological structures was significantly higher compared to fresh sperm, both in the breeding and non-breeding seasons (p ≤ 0.05). After the cryopreservation, the length and width of the head decreased significantly (p ≤ 0.05) compared to the corresponding sperm sizes before the cryopreservation. Analyzing the morphological char- acteristics of the tail part, we revealed the fact of its reduction after the cryopreservation, probably due to twisting, loop formation, and detachment of its part. Conclusions. It was found that in the non-breeding season, the number of spermatozoa with damaged morphological structures increases, compared to the breeding season, which must be taken into account when using reproductive technologies. The morphological and morphometric characteristics of Saanen buck spermatozoa change after the cryopreservation, regardless of the season, which indicates the need to choose the fertilization tactics that will ensure the selection of spermatozoa without any morphological abnormalities.

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Semen cryopreservation and storage in liquid nitrogen: Impact on chromatin compaction

Tamburrino,

Traini,

Ragosta

et al. 2024

Andrology

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BackgroundSperm cryopreservation is a consolidate option for long‐term male fertility preservation. The freezing/thawing procedure causes detrimental effects to spermatozoa, including damage to viability, motility, membrane composition, and DNA, whereas the effect on sperm chromatin compaction is less studied.ObjectivesThe primary aim of this study was to investigate the impact of cryopreservation on sperm chromatin compaction. Furthermore, the effect of cryopreservation on sperm parameters (motility, viability, chromatin compaction, and DNA fragmentation) was also assessed in relation to the storage time in liquid nitrogen.Materials and MethodsSemen samples, collected from 126 (92 normozoospermic and 34 oligozoospermic) patients undergoing routine semen analysis in the Andrology Laboratory of Careggi University Hospital of Florence, were frozen by conventional fast vapor freezing method. Sperm motility, viability, kinematic parameters (by computer‐aided sperm analysis [CASA]), chromatin compaction (by staining with both aniline blue [AB] and Chromomycin A3 [CMA3]), and sperm DNA fragmentation (sDF, by TUNEL/Propidium Iodide [PI]) were evaluated before freezing and after thawing at different timepoints.ResultsAfter 7 days of storage, a significant decline in sperm motility, viability, and kinematics parameters, as well as a significant increase in the percentage of sperm positivity to CMA3, AB, and sDF, were observed. It is noteworthy that while motility and viability decreased in almost all subjects, the increase in CMA3 and AB positivity was observed in 68.0% and 79.2% of samples, respectively. A progressive deterioration of sperm motility and viability, less evident for chromatin structure, was observed at longer times of storage (28 and 180 days).DiscussionOur results indicate that freezing/thawing procedures can alter chromatin structure. A reduction in protamine content and/or a modification in chromatin assembly can be hypothesized. Furthermore, the length of storage in liquid nitrogen appears to progressively affect sperm parameters, although it should be confirmed in larger cohort of subjects.ConclusionCurrent sperm cryopreservation protocols need to be improved with new strategies and personalized procedures aimed to minimize the damage.

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Sperm Cryopreservation

Cited by 3 publications

References 112 publications

Biomaterials for Testicular Bioengineering: How far have we come and where do we have to go?

Biomaterials for Testicular Bioengineering: How far have we come and where do we have to go?

Morphological and morphometric characteristics of fresh and cryopreserved spermatozoa of saanen bucks

Semen cryopreservation and storage in liquid nitrogen: Impact on chromatin compaction

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