Boar spermatozoa cryopreservation in low glycerol/trehalose enriched freezing media improves cellular integrity

Gutiérrez-Pérez, Oscar; Juárez-Mosqueda, Marı́a de Lourdes; Uribe‐Carvajal, Salvador; Ortega, María Elena Trujillo

doi:10.1016/j.cryobiol.2009.02.003

Cited by 79 publications

(53 citation statements)

References 54 publications

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“…The optimal concentrations of trehalose in the diluting solution and glycerol in the freezing solution were 200 mM and 2% (v/v), which differed, to some extent, from the findings of gutierrez-Perez et al [12] but led to slightly higher motility after freezing and thawing. The optimal equilibration time was 3 h, similar to the findings of Guthrie and Welch [11] but different from those of Pursel et al [31].…”

Section: Optimization and Screening Of Cryopreservation Schemes For Scontrasting

confidence: 50%

“…Jiang et al [17] used 9% LDL and 2% glycerol in combination and obtained a motility of 49.33% for frozen-thawed spermatozoa; gutierrez-Perez et al [12] obtained a motility of 42.25% using 250 mM trehalose and 1% glycerol in combination. The three substances protect the cell membrane by different but nonconflicting mechanisms.…”

Section: Effects Of the Combination Of Ldl Trehalose And Glycerol Omentioning

confidence: 99%

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A Study on Optimizing the Cryopreservation Methods for Bama Miniature Pig Semen

et al. 2012

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Abstract:In this study, the cryopreservation methods for Bama miniature pig semen were investigated and optimized. First, using an orthogonal experimental design, the semen freezing-thawing procedure for Bama miniature boars was optimized based on analysis of the effects of concentrations of LDL (LC, parameter A), trehalose (TC, parameter B) and glycerol (GC, parameter C), the equilibration time at 15°C (ET, parameter D), and the thawing method (TM, parameter E) on sperm motility. The results showed that the effects of the parameters could be arranged as A>C>B>D>E. The LDL concentration and final glycerol concentration had exceedingly significant effects on the motility of thawed spermatozoa (P<0.01), and the effects of the trehalose concentration, equilibration time at 15°C, and the thawing method were not significant (P>0.05). Scheme 2 (A 3 B 4 C 2 D 3 E 1 ) gave a motility of 52.26% after thawing. Then, using sperm motility, acrosome integrity, plasma membrane integrity, and DNA injury rate as indicators, four combinations, on the basis of scheme 2, were designed to analyze the protective effects of different combinations of LDL, glycerol, and trehalose; the results showed that combination of 9% LDL, 200 mM trehalose, and 2% glycerol (i.e., combination 4) demonstrated significantly better protective effects than the other combinations (P<0.05), further verifying that scheme 2 was the best for cryopreservation of Bama miniature boar semen. In this way, a method with favorable performance was established for cryopreservation of semen of Bama miniature boars.

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Section: Optimization and Screening Of Cryopreservation Schemes For Scontrasting

confidence: 50%

Section: Effects Of the Combination Of Ldl Trehalose And Glycerol Omentioning

confidence: 99%

A Study on Optimizing the Cryopreservation Methods for Bama Miniature Pig Semen

et al. 2012

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“…Furthermore, it protects the boar sperm from dehydration and intracellular ice formation during the semen freezing process [27]. Different types of sugar, e.g., trehalose, lactose, fructose, have been used in the freezing extender of boar semen [15,18].Recently, special interest on researches concerning with the energy utilization of the boar sperm have been noticed [13,15,18,20]. In general, the boar sperm is able to utilize a variety of substrates to obtain energy.…”

mentioning

confidence: 99%

“…Recently, special interest on researches concerning with the energy utilization of the boar sperm have been noticed [13,15,18,20]. In general, the boar sperm is able to utilize a variety of substrates to obtain energy.…”

mentioning

confidence: 99%

Cryopreservation of Boar Semen by Egg Yolk-Based Extenders Containing Lactose or Fructose is Better Than Sorbitol

2012

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ABSTRACT. The present study determined the effect of different types of sugars (lactose, fructose, glucose and sorbitol) used in egg yolkbased extender on the post-thawed boar semen quality. Twenty-two ejaculates from 6 fertility-proven Yorkshire boars were cryopreserved by liquid nitrogen vapor method. Sperm motility, viability, acrosome integrity and intact functional plasma membrane were determined at 0, 2 and 4 hr after thawing. It was found that the lactose-based extender resulted in a higher percentage of post-thawed sperm motility, viability, intact acrosome and functional plasma membrane than sorbitol-based extender (P<0.05) and fructose-based extender yielded a higher post-thawed sperm motility and viability than sorbitol-based extender (P<0.05). It could be concluded that sorbitol was not an effective sugar for the cryopreservation in boar semen. Boar semen cryopreservation greatly facilitates the distribution of valuable and desirable genes because the frozen semen can be transported over a long distance; and a stock of frozen semen enables the use of boar that is dead or ill [3]. However, the use of frozen-thawed (FT) boar semen is limited due to the low fertility outcomes compared to the extended fresh semen [9]. During cryopreservation, both physical and chemical factors including the rapid change in temperature or thermal stress, oxidative stress and osmotic stress lead to the damage of the sperm plasma membrane [19]. In general, the egg yolk-based freezing extender of boar semen consists of cryoprotectant, sugar, buffer and antibiotic [16]. Glycerol is the most common cryoprotectant used in the egg yolk-based semen extender for cryopreservation of the boar semen [14] and egg yolk is regarded as a non-permeable cryoprotectant. Sugar is mainly an energy source of the boar sperm. Furthermore, it protects the boar sperm from dehydration and intracellular ice formation during the semen freezing process [27]. Different types of sugar, e.g., trehalose, lactose, fructose, have been used in the freezing extender of boar semen [15,18].Recently, special interest on researches concerning with the energy utilization of the boar sperm have been noticed [13,15,18,20]. In general, the boar sperm is able to utilize a variety of substrates to obtain energy. Generally, monosaccharides are more common energy source used in extended fresh boar semen than disaccharides [20]. However, lactose (disaccharide) is commonly used for the cryopreservation of boar semen by many researchers [8,9,16,17]. In our previous studies [10,17] [24]. The stallion's cryopreserved sperms utilize sorbitol in the semen extender better than fructose and glucose [24]. In the boar, information associated with the effect of sugar types used in egg yolk-based semen extender on the FT semen qualities is limited. Therefore, the objective of the present study was to determine the effect of different sugar types (lactose, fructose, glucose and sorbitol) used in egg yolk-based semen extender on the sperm motility, sperm viability, acrosome integrity a...

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“…Buhr et al (2001) mostraram que nenhuma concentração de glicerol maximiza os parâmetros funcionais do esperma suíno criopreservado. Desta forma busca-se encontrar crioprotetores para a substituição do glicerol, a exemplo disso, a trealose (HU et al, 2009;GUTIÉRREZ-PÉREZ et al, 2009) e a dimetilacetamida (BIANCHI et al, 2008) mostram resultados promissores para a espécie suína, sugerindo desta forma sua utilização.…”

Section: Glicerolunclassified

Differences of Swine Sperm Cell and Cryoprotectants: Review

Domenico

Pedroso²,

Teixeira

2015

Nucleus Animalium

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RESUMO: Na espécie suína o emprego de sêmen congelado leva a índices reprodutivos insatisfatórios, quando comparados aos obtidos com sêmen refrigerado em inseminação artificial, pois o processo de criopreservação produz danos celulares conhecidos como crioinjúrias, que afetam a qualidade estrutural e funcional dos espermatozoides. As lesões ocasionadas têm sido atribuídas à mudança de temperatura, toxicidade de crioprotetores e estresse osmótico. Além das alterações da membrana plasmática que levam à maior sensibilidade da célula ao processo de criopreservação, ainda possui uma baixa defesa antioxidante citoplasmática e no plasma seminal favorecendo a danos oxidativos in vivo. Para as células sobreviverem ao processo de criopreservação é necessário a presença de agentes crioprotetores durante o resfriamento, congelamento e descongelamento. Os agentes crioprotetores são divididos entre externos ou não penetrantes e internos ou penetrantes.Palavras-chave: Inseminação artificial. Criopreservação. Crioprotetores. DIFFERENCES OF SWINE SPERM CELL AND CRYOPROTECTANTS: REVIEW SUMMARY:In swine the use of frozen semen leads to poor reproductive rates compared to those obtained with cooled semen in artificial insemination because the cryopreservation process produces cellular damage known as cryoinjuries, affecting the structural and functional quality of sperm. The injuries caused have been attributed to temperature change, cryoprotectants toxicity and osmotic stress. Besides the changes of the plasma membrane leading to increased sensitivity of the cell to the cryopreservation process, still has a low antioxidant defense cytoplasmic and seminal plasma favoring oxidative damage in vivo. For cells survive the cryopreservation process cryoprotectors agents during cooling, freezing and thawing is necessary. The cryoprotective agent are divided between external or internal or non-penetrating and penetrating.Keywords: Artificial insemination. Cryopreservation. Cryoprotectors. INTRODUÇÃOA suinocultura brasileira, a exemplo de outras cadeias produtivas do agronegócio, cresceu significativamente nos últimos anos. Esse crescimento deve-se principalmente ao melhoramento nutricional e genético dos animais. A base do melhoramento genético em suínos está relacionada à inseminação artificial, porém nesta espécie essa biotecnologia é apenas viável com sêmen refrigerado (MAPA, 2014). No qual possui algumas desvantagens em relação ao sêmen congelado, como o curto período de utilização do sêmen, a inviabilização da criação de bancos de sêmen e do comercio internacional de doses (ALONSO, 2009).A conservação do sêmen suíno através do congelamento, sem alteração dos resultados de fertilidade, é um dos objetivos da pesquisa na área da reprodução (FERNÁNDEZ-SANTOS et

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Boar spermatozoa cryopreservation in low glycerol/trehalose enriched freezing media improves cellular integrity

Cited by 79 publications

References 54 publications

A Study on Optimizing the Cryopreservation Methods for Bama Miniature Pig Semen

A Study on Optimizing the Cryopreservation Methods for Bama Miniature Pig Semen

Cryopreservation of Boar Semen by Egg Yolk-Based Extenders Containing Lactose or Fructose is Better Than Sorbitol

Differences of Swine Sperm Cell and Cryoprotectants: Review

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