Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects

Lê, Minh Tâm; Nguyen, Thi Thai Thanh; Nguyen, Thanh Tung; Nguyen, Van Toan; Nguyen, Thi Tam An; Nguyen, Vu Quoc Huy; Cao, Ngọc Thành

doi:10.1016/j.ejogrb.2019.01.001

Cited by 46 publications

(37 citation statements)

References 24 publications

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“…In a study by Le et al [34], morphology in spermatozoa was better preserved after devitrification with less defects observed when compared to conventional freezing. In our study, morphological changes in spermatozoa increased during vitrification and storage at the cost of intact acrosomes and sperm tails.…”

Section: Discussionmentioning

confidence: 95%

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: 95%

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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“…Pabon et al warmed vitrified human spermatozoa micropills (5-10 μl each) in 500 μl medium prewarmed and maintained at 44°C for 5 s and observed descent postthaw motility and mitochondrial activity [33]. Another report noted above warmed vitrified human sperm samples (30 μl pellets) in a 37°C water bath for 5 min resulting in motile and fertile sperm [32], indicating the flexibility of warming procedure.…”

Section: Vitrified Specimen Warmingmentioning

confidence: 98%

“…With 33 human semen samples, an experiment-controlled study found both slow cryopreservation and vitrification had similar results, but the latter was faster, easier and associated with lower toxicity and cost [28]. Le et al recently carried out a direct comparison between vitrification and slow cryopreservation [32]. They used 105 human fresh semen samples, exclusive of cryptozoospermia and azoospermia, divided them into washed and unwashed halves, and each group was split into two aliquots: one group cryopreserved by conventional freezing while the other by vitrification.…”

Section: Is Vitrification Superior To Conventional Freezing?mentioning

confidence: 99%

“…They used 105 human fresh semen samples, exclusive of cryptozoospermia and azoospermia, divided them into washed and unwashed halves, and each group was split into two aliquots: one group cryopreserved by conventional freezing while the other by vitrification. It was demonstrated that conventional freezing resulted in higher motility and viability, while spermatozoa undergoing vitrification were healthier regarding morphology and had fewer defects of sperm head, midpiece and tail [32]. Very recently, Pabon et al [33] used 47 human sperm samples to compare the efficiency of vitrification and conventional freezing protocols.…”

Section: Is Vitrification Superior To Conventional Freezing?mentioning

confidence: 99%

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Human sperm vitrification: the state of the art

Tao

Sanger

Saewu

et al. 2020

Reprod Biol Endocrinol

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Sperm cryopreservation has been widely used in assisted reproductive technology (ART) and has resulted in millions of live births. Two principal approaches have been adopted: conventional (slow) freezing and vitrification. As a traditional technique, slow freezing has been successfully employed and widely used at ART clinics whereas the latter, a process to solidify liquid into an amorphous or glassy state, may become a faster alternative method of sperm cryopreservation with significant benefits in regard to simple equipment and applicability to fertility centers. Sperm vitrification has its own limitations. Firstly, small volume of load is usually plunged to liquid nitrogen to achieve high cooling rate, which makes large volume sample cryopreservation less feasible. Secondly, direct contact with liquid nitrogen increases the potential risk of contamination. Recently, new carriers have been developed to facilitate improved control over the volume and speed, and new strategies have been implemented to minimize the contamination risk. In summary, although sperm vitrification has not yet been applied in routine sperm cryopreservation, its potential as a standard procedure is growing.

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“…The technique has already been widely explored and found its uses on embryos and oocytes and lately also for spermatozoa. [16][17][18][19][20][21]. Different studies have described individual protocols using different carriers and methods for standardization of the vitrification procedure.…”

Section: Techniques With Modified Protocolsmentioning

confidence: 99%

Sperm Cryopreservation: Principles and Biology

2020

J. Infertil. Reprod. Biol.

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Cryopreservation is a widely used method to preserve sperm prior to any cytotoxic therapy or testicular surgical intervention to be used later for assisted conception. Despite of multiple modifications in cryopreservation protocols, the yield of post-thaw good quality sperm has not been improved much. There is little data regarding factors affecting cryopreservation techniques and outcomes. Present review focuses on the basic biology of cryopreservation, its current protocols and effects on sperm proteomic and epigenomic modifications.

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Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects

Cited by 46 publications

References 24 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

Human sperm vitrification: the state of the art

Sperm Cryopreservation: Principles and Biology

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