Effect of glycerol, n, n-dimethylformamide and n-methyl-2-pyrrolidone on rabbit sperm stored at 4 °C and 16 °C

Domingo, Paula; Olaciregui, Maite; González, Noelia; Blás, Ignacio de; Gil, L.

doi:10.21451/1984-3143-ar2018-0100

Cited by 4 publications

(3 citation statements)

References 26 publications

(40 reference statements)

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“…In BoviFree ® and BioXcell ® , the rapid decrease in sperm motility and disruptions in plasma and acrosomal membrane integrity could also be caused by the presence of glycerol. This compound exerts a positive effect on cryopreserved spermatozoa [ 53 , 54 ], but it can damage cell membranes during liquid storage [ 55 ]. In spermatozoa, glycerol can damage various cell structures by inducing osmotic and non-osmotic changes [ 56 , 57 , 58 ].…”

Section: Discussionmentioning

confidence: 99%

The Effect of Different Extenders on the Quality Characteristics of European Red Deer Epididymal Sperm Stored at 5 °C

Dziekońska

Neuman

Burdal

et al. 2022

Animals

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The aim of this study was to evaluate the effect of different extenders on the quality of European red deer epididymal sperm stored at 5 °C. Epididymal spermatozoa were collected post mortem from 10 stags and diluted with three extenders (Bovidyl®, BoviFree®, and BioXcell®) and stored at 5 °C. Sperm motility (TMOT), motility parameters (system CASA), plasma membrane integrity (SYBR-14+/PI−), acrosomal membrane integrity (FITC-PNA−/PI−), mitochondrial activity (JC-1/PI), viability, and apoptotic-like changes (YOPRO/PI) were evaluated. The analyses were conducted on the first and successive days of storage (D1–D7). The applied extender, storage time, and the interactions between these factors significantly (p < 0.001) affected most of the analyzed parameters whose values were highest in sperm samples stored in Bovidyl®, regardless of storage time. In Bovidyl®, BoviFree®, and BioXcell® extenders, TMOT values were estimated at 83%, 63%, and 59%, respectively, on D3. The extenders significantly influenced DNA integrity on D7. The percentage of dead sperm increased from D3. The quality of stored sperm cells was significantly influenced by the extenders’ biochemical composition. BoviFree® and BioXcell® contain glycerol which could contribute to deteriorating the quality of spermatozoa stored at 5 °C. Sperm cells stored in the egg yolk-based extender (Bovidyl®) were characterized by the highest viability and functionality.

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Section: Discussionmentioning

confidence: 99%

The Effect of Different Extenders on the Quality Characteristics of European Red Deer Epididymal Sperm Stored at 5 °C

Dziekońska

Neuman

Burdal

et al. 2022

Animals

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“…Recently, Domingo et al, [105] studied comparison of different semen extenders and cryoprotectant agents to enhance cryopreservation of rabbit spermatozoa and found that the addition of dimethylformamide (DMF) to INRA 96® exerts a protective effect on the membrane of spermatozoa improving seminal quality [105]. Although many efforts have been made to optimize cryopreservation extenders and protocols for rabbit sperm, many questions remain unanswered.…”

Section: Ratmentioning

confidence: 99%

Cryopreservation Methods and Frontiers in the Art of Freezing Life in Animal Models

Aljaser¹

2022

Veterinary Medicine and Science

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The development in cryobiology in animal breeding had revolutionized the field of reproductive medicine. The main objective to preserve animal germplasm stems from variety of reasons such as conservation of endangered animal species, animal diversity, and an increased demand of animal models and/or genetically modified animals for research involving animal and human diseases. Cryopreservation has emerged as promising technique for fertility preservation and assisted reproduction techniques (ART) for production of animal breeds and genetically engineered animal species for research. Slow rate freezing and rapid freezing/vitrification are the two main methods of cryopreservation. Slow freezing is characterized by the phase transition (liquid turning into solid) when reducing the temperature below freezing point. Vitrification, on the other hand, is a phenomenon in which liquid solidifies without the formation of ice crystals, thus the process is referred to as a glass transition or ice-free cryopreservation. The vitrification protocol applies high concentrations of cryoprotective agents (CPA) used to avoid cryoinjury. This chapter provides a brief overview of fundamentals of cryopreservation and established methods adopted in cryopreservation. Strategies involved in cryopreserving germ cells (sperm and egg freezing) are included in this chapter. Last section describes the frontiers and advancement of cryopreservation in some of the important animal models like rodents (mouse and rats) and in few large animals (sheep, cow etc).

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“…Despite the fact that glycerol has become a popular CPA for cryopreservation of almost all domestic animals' sperm, it is not suitable for rabbit sperm due to poor sperm plasma membrane permeability and a high activation energy requirement. (Domingo et al, 2019). On the other hand, studies have shown that dimethyl sulfoxide (DMSO) and/or amides are the cryoprotectants of choice for rabbit semen (Mocé et al, 2010), as they can stabilise the sperm membrane during freezing/thawing processes (Mocé and Vicente, 2009).…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of rabbit semen: impacts of permeable and non-permeable mixture of cryoprotectant, male group individuality, freezing rate, semen package size and antioxidant bovine serum albumin on rabbit semen freezability

et al. 2022

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In the present study, three experiments were designed to identify the most appropriate technique for freezing rabbit semen. Experiment 1 aimed to determine the optimal levels of dimethyl sulfoxide (DMSO) contents in freezing medium and their effects on individual bucks. Semen ejaculates for each buck (n=15 bucks) were mixed and split into three portions for extension with a freezing medium containing varying concentrations of DMSO (0.75, 1.0, and 1.4 M). Diluted semen samples were packaged in 0.25 mL straws and suspended above liquid nitrogen (LN) for 10 min, then dipped in LN. A few days after freezing, post-thaw semen evaluation was assessed, and according to the results, six bucks and an extender containing 0.75 M of DMSO were used for experiments 2 and 3. In experiment 2, the pooled semen from 6 bucks was divided into two portions for packaging in two straw sizes (0.25 and 0.50 mL). Each straw size was divided into five groups and suspended at different heights above LN (2, 4, 6, 8, and 10 cm) for 10 minutes before being preserved in LN. In experiment 3, the pooled semen was divided into four portions for dilution with freezing medium containing different concentrations of bovine serum albumin (BSA; 0, 2.5, 5.0, and 7.5 mg/mL). Semen samples were packaged in a 0.50 mL straw and suspended 10 min, 4 cm above LN for freezing. Pre-freezing and post-thawing, semen samples were evaluated for semen quality. Results showed that the extender containing 0.75 M DMSO had higher significant values for post-thaw sperm motility, longevity, acrosome integrity and sperm plasma membrane permeability. Bucks’ individuality had significant effects on post-thaw motility, acrosome and sperm plasma membrane integrity. A significant interaction was recorded between DMSO concentrations and bucks’ individuality on sperm longevity. Semen package sizes had no significant effects on the evaluated parameters. Semen was frozen at 2 and 4 cm above LN had significantly better post-thaw quality. BSA at concentrations 5 and 7.5 mg/mL improved recovery rates of acrosome integrity and sperm membrane permeability. DMSO 0.75 M and freezing 4 cm above LN seem to be more adequate for rabbit semen cryopreservation. The appropriate level of DMSO differs between bucks, as the post-thaw sperm longevity is affected. BSA enhanced acrosome and sperm membrane integrity. Results obtained will need further investigation to be confirmed in the field.

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Effect of glycerol, n, n-dimethylformamide and n-methyl-2-pyrrolidone on rabbit sperm stored at 4 °C and 16 °C

Cited by 4 publications

References 26 publications

The Effect of Different Extenders on the Quality Characteristics of European Red Deer Epididymal Sperm Stored at 5 °C

The Effect of Different Extenders on the Quality Characteristics of European Red Deer Epididymal Sperm Stored at 5 °C

Cryopreservation Methods and Frontiers in the Art of Freezing Life in Animal Models

Cryopreservation of rabbit semen: impacts of permeable and non-permeable mixture of cryoprotectant, male group individuality, freezing rate, semen package size and antioxidant bovine serum albumin on rabbit semen freezability

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