Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification

Pradieé, J.; Esteso, M.C.; Castaño, C.; Toledano-Dı́az, A.; Guerra, R.; Santiago-Moreno, J.

doi:10.1111/and.12629

Cited by 33 publications

(31 citation statements)

References 34 publications

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“…In humans, sucrose alone adequately preserves semen viability and motility [24]. Sucrose alone has also been demonstrated to support successful vitrification and ultimately fertilization in wild goats [23], mouflons [10], and stallions [25]. However, our results are in agreement with other studies in which some protein source is necessary to support survival during the initial cold shock of cryopreservation.…”

Section: Discussionsupporting

confidence: 90%

“…One way of avoiding damage caused by crystal formation is semen vitrification [5], which has been studied in different animal species and in humans as an alternative to conventional freezing [1,[5][6][7][8][9][10]. Vitrification has been used widely for embryo storage [3] and has been a scope of research in the last decade for sperm cryopreservation [11].…”

Section: Introductionmentioning

confidence: 99%

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Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function

Pipan

Casal

Šterbenc

et al. 2020

Animals

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Simple Summary: Soy lecithin and sucrose were used at different concentrations to develop and compare different vitrification methods for the cryopreservation of canine semen. All of the sperm quality characteristics were effectively preserved after devitrification when vitrification extenders containing soy lecithin at 1% and 0.25 M sucrose were used. The results suggest that vitrification is effective, fast, and simple for cryopreservation of canine semen. Furthermore, the use of soy lecithin in lieu of animal proteins (e.g., egg yolk) facilitates semen shipping to countries with strict import requirements.Abstract: A challenge in freezing semen for short and long-term availability is avoiding damage to intact spermatozoa caused by the freezing process. Vitrification protocols provide better results through less manipulation of semen and shorter freezing time compared to slow freezing techniques. Our research was aimed at improving vitrification methods for canine semen. Semen quality was determined in 20 ejaculates after collection. Each ejaculate was divided into eight aliquots, each with a different extender. The control extender contained TRIS, citric acid, fructose, and antibiotics. Soy lecithin and sucrose were added to the control extender at different concentrations to make up the test extenders and final concentration of 50 × 10 6 spermatozoa/mL. From each group, a 33 µL (1.65 × 10 6 spermatozoa) suspension of spermatozoa was dropped directly into liquid nitrogen and devitrified at least one week later and evaluated as before. Soy lecithin at 1% and 0.25 M sucrose added to the base vitrification media effectively preserved all sperm qualities. Our results demonstrate the effectiveness of our methods. Vitrification media containing sucrose and soy lecithin cause a minimal decline in quality of canine semen after devitrification. Furthermore, extenders used in our research did not contain egg yolk, which was replaced by soy lecithin, thus allowing for ease of shipping to other countries with strict requirements.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function

Pipan

Casal

Šterbenc

et al. 2020

Animals

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“…However, vitrified ram sperm had a lower quality than sperm cryopreserved by conventional method. A similar situation was reported in mouflon sperm vitrified with sucrose [160]. On the contrary, vitrification of Iberian Ibex (Capra pyrenaica) sperm yielded a similar post-thaw quality and in vitro fertilization rate than sperm cryopreserved by conventional method [161].…”

Section: Molecular Aspects Of Those Novel Strategies To Reduce Sperm supporting

confidence: 75%

Sperm Cryodamage in Ruminants: Understanding the Molecular Changes Induced by the Cryopreservation Process to Optimize Sperm Quality

Peris-Frau¹,

Soler²,

Iniesta‐Cuerda³

et al. 2020

IJMS

139

118

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Sperm cryopreservation represents a powerful tool for livestock breeding. Several efforts have been made to improve the efficiency of sperm cryopreservation in different ruminant species. However, a significant amount of sperm still suffers considerable cryodamage, which may affect sperm quality and fertility. Recently, the use of different “omics” technologies in sperm cryobiology, especially proteomics studies, has led to a better understanding of the molecular modifications induced by sperm cryopreservation, facilitating the identification of different freezability biomarkers and certain proteins that can be added before cryopreservation to enhance sperm cryosurvival. This review provides an updated overview of the molecular mechanisms involved in sperm cryodamage, which are in part responsible for the structural, functional and fertility changes observed in frozen–thawed ruminant sperm. Moreover, the molecular basis of those factors that can affect the sperm freezing resilience of different ruminant species is also discussed as well as the molecular aspects of those novel strategies that have been developed to reduce sperm cryodamage, including new cryoprotectants, antioxidants, proteins, nanoparticles and vitrification.

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“…Some non-permeable additives with cryoprotective activity, such as human serum albumin and sucrose, have been successfully used in the kinetic vitrification of human sperm [16]. This modified method was subsequently used to successfully cryopreserve dog [17] and fish sperm [18], and more recently Iberian ibex (Capra pyrenaica) [6] and European mouflon (Ovis musimon) [19] sperm. Vitrification occurs-or not-depending on the cooling rate, the viscosity of the solution, and the volume to be preserved [20].…”

Section: Introductionmentioning

confidence: 99%

“…The plasmalemma and acrosomal membranes are major sites of cryopreservation-induced sperm damage [24]; this is certainly true for wild ruminant sperm vitrified in pellets [19]. The size of the sperm head can also be affected.…”

Section: Introductionmentioning

confidence: 99%

Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages

et al. 2020

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Sperm cryopreservation by ultra-rapid cooling based on dropping small volumes of sperm suspension directly into liquid nitrogen, has been successful in some wild ruminant species, including the Iberian ibex (Capra pyrenaica). In ultra-rapid cooling, the contents of these droplets are expected to enter a stable, glass-like state, but to the best of our knowledge no information exists regarding the presence or absence of ice formation in the extracellular milieu when using this technique. Different modifications to the extracellular milieu likely inflict different types of damage on the plasmalemma, the acrosome and mitochondrial membranes. The aims of the present work were: 1) to examine the physical state of the extracellular milieu after cryopreservation at slow and ultra-rapid cooling rates-and thus determine whether ultra-rapid cooling vitrifies the extracellular milieu; and 2) to compare, using conventional sperm analysis techniques and scanning and transmission electron microscopy, the damage to sperm caused by these two methods. Sperm samples were obtained by the transrectal ultrasound-guided massage method (TUMASG) from anesthetized Iberian ibexes, and cryopreserved using slow and ultra-rapid cooling techniques. Sperm motility (22.95 ± 3.22% vs 4.42 ± 0.86%), viability (25.64 ± 3.71% vs 12.8 ± 2.50%), acrosome integrity (41.45± 3.73% vs 27.00 ± 1.84%) and mitochondrial membrane integrity (16.52 ± 3.75% vs 4.00 ± 0.65%) were better after slow cooling (P<0.001) than after ultrarapid technique. Cryo-scanning electron microscopy (Cryo-SEM) suggested that the vitrified state was not achieved by ultra-rapid cooling, and that the ice crystals formed were smaller and had more stretchmarks (P<0.001) than after slow cooling. Scanning electron microscopy revealed no differences in the types of damage caused by the examined techniques, although transmission electron microscopy showed the damage to the plasmalemma and mitochondrial membrane to be worse after ultra-rapid cooling. In conclusion ultra-rapid cooling provoked more membrane damage than slow cooling, perhaps due to the extracellular ice crystals formed.

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Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification

Cited by 33 publications

References 34 publications

Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function

Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function

Sperm Cryodamage in Ruminants: Understanding the Molecular Changes Induced by the Cryopreservation Process to Optimize Sperm Quality

Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages

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