Evaluation of l-glutamine for cryopreservation of boar spermatozoa

Mercado, E. de; Hernández, M.; Sanz, E.; Rodríguez, A.; Gómez, E.; Vázquez, J.M.; Rodríguez‐Martínez, H.; Roca, Jordi

doi:10.1016/j.anireprosci.2008.11.014

Cited by 40 publications

(18 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Many substances have been tested to determine whether they enhance the resistance of spermatozoa to the stress of freezing and thawing in several species, including the pig (Roca et al 2004, 2005b, Breininger et al 2005, Bathgate et al 2006, Corcuera et al 2007, De Mercado et al 2009, Hu et al 2009). Pentoxifylline, a methylxanthine derivate, is an inhibitor of phosphodiesterase.…”

Section: Introductionmentioning

confidence: 99%

Pentoxifylline added to freezing or post-thaw extenders does not improve the survival or in vitro fertilising capacity of boar spermatozoa

Gil¹,

Hernández²,

Roca³

et al. 2010

Reproduction

View full text Add to dashboard Cite

This study evaluated whether pentoxifylline added to freezing and thawing extenders influenced the function of boar spermatozoa. In Experiment 1, pooled ejaculated sperm-rich fractions were frozen in 0.5 ml straws after dilution in extender supplemented with pentoxifylline to a final concentration of 0, 2, 4, 8, 16 or 32 mM. The addition of 4, 8, 16 and 32 mM pentoxifylline to the freezing extender significantly decreased the progressive and total motility of spermatozoa. The percentage of viable spermatozoa with intact acrosomes as well as the penetration rate and the efficiency of fertilisation were significantly lower in pentoxifylline-treated groups compared with the untreated control. In Experiment 2, a pool of three straws with 'good' post-thaw sperm quality parameters and another three straws with 'poor' sperm quality were diluted in extender with 0, 1, 2, 4, 8, 16 or 32 mM pentoxifylline. Post-thaw samples with both 'good' and 'poor' sperm quality with 0, 2, 4, 8 and 16 mM were used to assess IVF parameters. The addition of pentoxifylline to post-thaw extender did not improve the post-thaw motility or viability of spermatozoa compared with the control. The in vitro penetration was higher (P!0.05) than the control for oocytes fertilised with spermatozoa that were thawed and incubated in extender with 4, 8 and 16 mM pentoxifylline. However, no differences were observed in the efficiency of fertilisation. We conclude that pentoxifylline, as a supplement added to the freezing extender, has a deleterious effect and that it does not improve the survival or in vitro fertilising efficiency of frozen-thawed boar spermatozoa when added after thawing.

show abstract

Section: Introductionmentioning

confidence: 99%

Pentoxifylline added to freezing or post-thaw extenders does not improve the survival or in vitro fertilising capacity of boar spermatozoa

Gil¹,

Hernández²,

Roca³

et al. 2010

Reproduction

View full text Add to dashboard Cite

show abstract

“…CFDA+/PI-stained sperm were classified as having an intact plasma-membrane. Sperm acrosomal-membrane integrity was evaluated using a FITC-peanut agglutinin (FITC-PNA; Sigma-Aldrich)/PI fluorescent staining following a modification [6,11] and analyzed using flow cytometry. Analyzed sperm were classified into the following categories: viable acrosome-intact (PNA-/PI-); viable acrosome-reacted (PNA+/PI-); dead acrosome-reacted (PNA+/PI+) and total acrosome-reacted (PNA+).…”

mentioning

confidence: 99%

Evaluation of Different Cryoprotectants (CPAs) in Boar Semen Cryopreservation

Kim

Lee

et al. 2011

J. Vet. Med. Sci.

View full text Add to dashboard Cite

ABSTRACT. This study was performed to evaluate the use of dimethylacetamide (DMA) and dimethyl sulfoxide (DMSO) in boar sperm cryopreservation. Semen from eight boars was cryopreserved following treatment with 3, 5, and 7% DMA and DMSO, and 3% glycerol (control). After thawing, sperm conventional parameters and membrane integrities were evaluated. There were no significant differences among different DMA concentrations in all evaluations. Membrane intactness were higher in 5% and 7% DMSO than 3% DMSO (P<0.05). Sperm motility of 5% DMSO was lower than that of 3% glycerol (P<0.005), and membrane intactness were lower in 5% DMA and DMSO than 3% glycerol (P<0.05). DMA and DMSO didn't improve sperm quality and glycerol remains the most useful for boar sperm cryopreservation. Glycerol (at a concentration of 3%) is the most commonly used cryoprotectant (CPA) for boar sperm [3], but it has toxic effects as well as contraceptive effects on sperm [8]. Its detrimental effects have stimulated the study of alternative CPAs such as N,N-dimethylacetamide (DMA) or dimethyl sulfoxide (DMSO) [2,10]. Due to its highly hydrophilic nature and low molecular weight, DMA reduces the formation of intracellular ice crystals and increases membrane permeability, thus decreasing osmotic damage [2]. The penetration of DMSO is also rapid, due to its lower molecular weight relative to glycerol [10] and DMSO has been used to successfully cryopreserve sperm [9]. However, there are few studies on the uses of DMA and DMSO for freezing boar semen. Therefore, the objective of this study was to evaluate the effectiveness of DMA and DMSO as possible replacement for glycerol in boar semen cryopreservation.Three CPAs were used in this study: glycerol, DMA, and DMSO (all from Sigma-Aldrich, St. Louis, MO, U.S.A.). Glycerol was used at a final concentration of 3% as a control, and the others at final concentrations of 3, 5, and 7%. We first compared the three concentrations of DMA and DMSO (n=5), and the concentrations showing the best result for each CPA were then compared with 3% glycerol (n=8).Ejaculate from 8 Duroc boars was collected and processed according to the straw freezing procedure [1] with some modification. Sperm-rich fractions were extended in Beltsville Thawing Solution (BTS) and were slowly cooled to 15°C for 3 hr. After centrifuging, the semen pellet was resuspended in lactose-egg yolk (LEY) extender. After further cooling to 5°C for 90 min, LEY-extended semen was aliquot for treatment with each CPAs and then two parts LEY-extended semen were mixed with one part freezing extenders (LEY extender containing 1.5% Equex STM [Nova Chemical Sales, Scituate Inc., MA, U.S.A.] and the CPAs described above, v/v). The semen was loaded into 0.5-ml straws (IMV, L'Aigle, France), and held in liquid nitrogen vapor 3 cm above the liquid for 20 min. The straws were then plunged into the liquid nitrogen. After thawing the straws at 37°C for 20 sec [5], thawed semen was evaluated for sperm motility [13], morphological defect, viability by eosin-nigrosin st...

show abstract

“…On the male, the efforts are centralized in the identification of differences between "good" and "bad" boar freezers (Watson, 1995;Thurston et al, 2001; and in the optimization of the cryopreservation protocols including new packaging systems (Eriksson and Rodriguez-Martinez, 2000), optimization of freezing and thawing velocities (Thurston et al, 2003; or modifications of freezing extenders (He et al, 2001, Roca et al, 2003, de Mercado et al, 2009). On the female, the effect of the timing of insemination with respect to the ovulation on fertilization rate has been studied (Bolarin et al, 2006;Larsson, 1976;.…”

Section: Applications and Limitations Of Frozen-thawed Sperm Dosesmentioning

confidence: 99%

“…To achieve these objectives, several fronts have been tackled to try to include most of the factors that may have an impact on the fertilizing ability of FT sperm (male, female and artificial insemination techniques). The strategies have focused on improving the artificial insemination (AI) technique (Roca et al, 2003), establishing optimal timing between insemination and ovulation , developing new packaging systems for the sperm (Eriksson et al, 2001), optimizing the freezing-thawing protocols or modifying the composition of the freezing diluents to reduce the damage during the cryopreservation process (de Mercado et al, 2009;Groβfeld et al, 2008;Yamaguchi et al, 2009). Although the advances towards increasing the fertilizing ability of FT sperm are remarkable, the use of this type of sperm commercially is still utopian.…”

Section: Literature Citedmentioning

confidence: 99%

Effect of the Pre-Freezing Treatment of Boar Sperm With Cholesterol-Loaded Cyclodextrins on the Functionality and the Fertilizing Ability of the Sperm After Cryopreservation

Almenar¹

View full text Add to dashboard Cite

Evaluation of l-glutamine for cryopreservation of boar spermatozoa

Cited by 40 publications

References 32 publications

Pentoxifylline added to freezing or post-thaw extenders does not improve the survival or in vitro fertilising capacity of boar spermatozoa

Pentoxifylline added to freezing or post-thaw extenders does not improve the survival or in vitro fertilising capacity of boar spermatozoa

Evaluation of Different Cryoprotectants (CPAs) in Boar Semen Cryopreservation

Effect of the Pre-Freezing Treatment of Boar Sperm With Cholesterol-Loaded Cyclodextrins on the Functionality and the Fertilizing Ability of the Sperm After Cryopreservation

Contact Info

Product

Resources

About