New permeable cryoprotectant-free vitrification method for native human sperm

Aizpurúa, Jon; Medrano, Llanos; Enciso, María; Sarasa, Jonás; Romero, Alejandro; Fernandez, Maria Aparecida; Gómez‐Torres, María José

doi:10.1093/humrep/dex281

Cited by 53 publications

(46 citation statements)

References 26 publications

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“…Vitrification has been successfully used in freezing of oocytes and embryos with the benefit of being a quick process that does not require expensive equipment [8]. Furthermore, by directly plunging samples into liquid nitrogen, solidification of living cells is achieved without intracellular ice crystallization [32]. However, spermatozoa are osmotically fragile cells and cannot withstand the lethal effect of permeable cryoprotectants at high concentrations needed for successful vitrification [22].…”

Section: Discussionmentioning

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: Discussionmentioning

confidence: 99%

“…This procedure was first described by Isachenko et al [23] for human semen and soon after also for canine sperm vitrification [8]. Recently, studies using human sperm vitrification protocols showed superior results in motility, viability, DNA structure, acrosome and morphology when compared to conventional freezing protocols [11,32,33].…”

Section: Discussionmentioning

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

View full text Add to dashboard Cite

show abstract

“…Recently published results from the use of the permeable cryoprotectant-free vitrification protocol (Easy-Sperm®, iGLS S.L., Alicante, Spain) on normozoospermic sperm samples have shown improved preservation of sperm structure and function compared to conventional slow freezing. Sperm motility, vitality, DNA integrity, acrosome and cytoskeleton structure were significantly better preserved in vitrified samples [1].…”

Section: Accepted Manuscriptmentioning

confidence: 93%

Section: Accepted Manuscriptmentioning

confidence: 99%

“…Recently, the development of a new permeable cryoprotectant-free vitrification protocol (Easysperm®, iGLS S.L., Alicante, Spain) for human spermatozoa provided an enhanced and reliable alternative for sperm cryopreservation [1]. This novel method has been tested in normozoospermic samples rendering better preservation of sperm quality on all the parameters studied (motility, vitality, DNA structure, acrosome reaction, morphology and cytoskeleton) compared to slow freezing [1]. At our centre (IVF-Spain Alicante, Spain) we have successfully used this protocol to cryopreserve and use for fertilisation a sperm sample from a couple whose female partner (39 years old) had ovarian failure and was recommended egg donation treatment with his partner´s (40 years old) sperm.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

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