Stallion sperm freezing with sucrose extenders: A strategy to avoid permeable cryoprotectants

Consuegra, C.; Crespo, F.; Bottrel, M.; Ortiz, Isabel; Dorado, J.; Díaz-Jiménez, María; Pereira, B.; Hidalgo, M.

doi:10.1016/j.anireprosci.2018.02.013

Cited by 25 publications

(15 citation statements)

References 46 publications

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“…Our results showed the extender containing only non‐permeable agents was able to protect plasma membrane and DNA integrity in both groups, as no differences were found in comparison to extender containing glycerol. In the same way, a combination of sucrose and BSA resulted in high plasma membrane and DNA integrity percentages after stallion sperm cryopreservation in previous reports (Consuegra et al., ; Hidalgo et al., ). Discrepancy in motility results between fertile and subfertile groups regarding the use of glycerol might be explained because the effectiveness of non‐permeable cryoprotectants could be subjected to the initial sperm quality of the sample.…”

Section: Discussionsupporting

confidence: 67%

“…However, permeable cryoprotectants used are known to exert a toxic effect in the sperm cell (Fahy, ), mostly due to the intrinsic toxicity and osmotic stress (Macías García et al., ). Recently though, sperm freezing avoiding permeable cryoprotectants has been successfully developed in fertile stallions (Consuegra et al., ) and donkeys (Diaz‐Jimenez et al., ). Additionally, the use of non‐permeable cryoprotectants has shown to provide protection against cryoinjuries without toxic effects (Swain & Smith, ).…”

Section: Introductionmentioning

confidence: 99%

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Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Díaz-Jiménez

Dorado

Pereira

et al. 2019

Reprod Domestic Animals

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Sperm from fertile donkeys have been successfully frozen in absence of permeable cryoprotectants. The aim of this study was to determine whether this cryopreservation method is suitable for subfertile donkeys in comparison to conventional sperm freezing with glycerol. Ejaculates were collected from four Andalusian Donkeys: three fertile and one subfertile. Semen was frozen with an extender containing glycerol (GLY), or adding instead sucrose 0.25 molar and 1% bovine serum albumin (SUC) as non-permeable cryoprotectants. After thawing, samples were assessed for total (TM, %) and progressive (PM, %) sperm motility by CASA, plasma membrane integrity (PMI, %) by epifluorescence microscopy and DNA integrity (DFI, %) by SCSA. Results (mean ± SD) were compared between extenders in fertile and subfertile donkeys using the Student's t test. No differences between GLY and SUC treatments were found in the fertile group for the sperm parameters assessed. In subfertile donkey ejaculates, GLY resulted in significantly higher values than SUC for TM (25.5 ± 3.1 vs.19.6 ± 1.9) and PM (13.3 ± 5.1 vs. 4.0 ± 1.2), respectively. In conclusion, considering all the sperm parameters assessed, sperm freezing in absence of permeable cryoprotectants may not be still an option for cryopreservation of subfertile donkey sperm. K E Y W O R D S cryoprotectant, donkey, fertility, freezing, sperm | 103 DIAZ-JIMENEZ Et Al. | 105 DIAZ-JIMENEZ Et Al. ACK N OWLED G M ENTS The authors are grateful to 'Centro Rural Malpica' for providing animals and facilities. This study was funded by MINECO grant AGL2013-42726-R. M. Diaz-Jimenez is supported by a FPU fellowship from Spanish MECD ('Ministerio de Educación, Cultura y Deporte', Spain). CO N FLI C T O F I NTE R E S T None of the authors have any conflict of interest to declare. AUTH O R CO NTR I B UTI O N S M Hidalgo and M Diaz-Jimenez contributed to all sections. J Dorado and I Ortiz contributed to the study design, preparation and final approval of the manuscript. C Consuegra and B Pereira performed the experiments. All the authors were involved in revision and approval of the final version of the manuscript.

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Section: Discussionsupporting

confidence: 67%

Section: Introductionmentioning

confidence: 99%

Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Díaz-Jiménez

Dorado

Pereira

et al. 2019

Reprod Domestic Animals

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“…Serum albumin has been traditionally added to the vitrification extender for sperm cryopreservation in several species. Thus, human serum albumin has been employed at 1% in human [29]; and BSA at 0.5% in rabbit [12]; 1% in equine [35], dog [19], fish [28], donkey [36] and wild goat [17]; and 2% in ram [37]. This molecule has shown to reduce oxidative stress [38] and to protect the sperm membrane against cryodamage, although the exact mechanism is still not clear [39].…”

Section: Discussionmentioning

confidence: 99%

Vitrification of Donkey Sperm: Is It Better Using Permeable Cryoprotectants?

Hidalgo

Díaz-Jiménez

Consuegra

et al. 2020

Animals

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Vitrification by direct exposure of sperm to liquid nitrogen is increasing in popularity as an alternative to conventional freezing. In this study, the effect of permeable cryoprotectant agents for donkey sperm vitrification was compared to an extender containing non-permeable cryoprotectants. First, three different concentrations of sucrose (0.1, 0.2, and 0.3 molar, M) and bovine serum albumin, BSA (1, 5, and 10%) were compared. Secondly, the concentration of non-permeable agents producing the most desirable results was compared to an extender containing glycerol as permeable agent. Vitrification was performed by dropping 30 μL of sperm suspension directly into LN2 and warming at 42 °C. Sperm motility (total, TM; and progressive, PM) and plasma membrane integrity, PMI (mean ± SEM) were statistically compared between treatments. Sucrose 0.1 M showed a significantly higher percentage of total sperm motility (21.67 ± 9.22%) than sucrose 0.2 M (14.16 ± 4.50%) and 0.3 M (8.58 ± 6.22%); and no differences were found in comparison to the control (19.71 ± 10.16%). Vitrification with sucrose 0.1 M or BSA 5% obtained similar results for TM (21.67 ± 9.22% vs. 19.93 ± 9.93%), PM (13.42 ± 6.85% vs. 12.54 ± 6.37%) and PMI (40.90 ± 13.51% vs. 37.09 ± 14.28); but both showed higher percentages than glycerol (TM = 9.71 ± 4.19%; PM = 5.47 ± 3.17%; PMI = 28.48 ± 15.55%). In conclusion, donkey sperm vitrification in spheres using non-permeable cryoprotectants exhibited better sperm motility and viability parameters after warming than sperm vitrification using extenders containing permeable cryoprotectants.

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“…In an appropriate concentration, sucrose can improve the cell dehydration and effectively reduce intracellular ice formation in a shorter period, but high sucrose concentration can reduced sperm motility because high viscosity of the medium might inhibit normal sperm motility [13].…”

Section: Discussionmentioning

confidence: 99%

DMSO and sucrose combination as a cryoprotectant of Najawa carp (Cyprinus carpio L.) sperm cryopreservation

2020

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This study was designed to evaluate a DMSO (Dimethyl Sulfoxide) and sucrose combination as a cryoprotectant for cryopreservation of Najawa carp (Cyprinus carpio L.) sperm. The experiment was conducted according to the Completely Randomized Design (CRD) with four treatments 10% DMSO + 0.2 M sucrose, 10% DMSO + 0.1 M sucrose, 5% DMSO + 0.2 M sucrose, 5% DMSO + 0.1 M sucrose. Each treatment consisted of three replications. The parameter observed were sperm progressive motility of fresh sperm, diluted sperm before freezing, and diluted sperm after freezing 1 day; 7 days; 14 days; 21 days; 28 days. The data were analyzed by ANOVA. The result showed that there was no significant difference between treatments (Sig. > 0.05). The highest relative protection was 89.51% carried out by 5% DMSO + 0.1 M sucrose.

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Stallion sperm freezing with sucrose extenders: A strategy to avoid permeable cryoprotectants

Cited by 25 publications

References 46 publications

Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Vitrification of Donkey Sperm: Is It Better Using Permeable Cryoprotectants?

DMSO and sucrose combination as a cryoprotectant of Najawa carp (Cyprinus carpio L.) sperm cryopreservation

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