Effects of Various Cryoprotective Agents and Membrane-Stabilizing Compounds on Bull Sperm Membrane Integrity after Cooling and Freezing

Leeuw, Frans L.; Leeuw, A.M. de; Daas, J.H.G. den; Colenbrander, B.; Verkleij, Arie J.

doi:10.1006/cryo.1993.1005

Cited by 170 publications

(97 citation statements)

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“…Extenders containing soybean lecithin could be an alternative to the conventional extenders that include egg yolk [5,18] . De Leeuw et al has noted that bull sperm survive freezing more effectively in egg yolk-containing diluents than in soybean lecithin [19] . In this study, the freeze-thaw process negatively affected sperm motility and acrosome integrity (P<0.001).…”

Section: Discussionmentioning

confidence: 99%

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Üstüner¹,

Alçay²,

Nur³

et al. 2014

Kafkas Univ Vet Fak Derg

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The aim of the current study was to evaluate the effect of egg yolk and different soybean lecithin concentrations on the efficiency of ram semen cryopreservation and to test the fertilizing ability of frozen-thawed ram semen. Ejaculates with a thick consistency, rapid wave motion (3-5), and >70% initial motility were pooled. Pooled semen were then divided into four groups and diluted at 1/5 (semen/extender) with 1%, 3%, 6% lecithin (L1, L3 and L6) or 20% egg yolk (EY20) using the two-step dilution method. As expected, the results of the current study showed that both motility and the rates of defective acrosomes in sperm were negatively affected by the cryopreservation procedure (P<0.001). The motility values of at 5°C and post-thawed semen in the EY20 group were significantly higher than those in the L1, L3 and L6 groups (P<0.05). There were no differences in motility rates among the lecithin groups at the dilution, cooling, equilibration or post thawing stages (P>0.05). The results of in vitro fertilization, as assessed by the rate of blastocyst formation, were more successful in the EY20 group than those noted in different lecithin groups. In conclusion, freezing ram semen with an extender containing egg yolk could yield better post-thaw sperm parameters and embryonic development compared to lecithin containing extenders. Keywords: Cryopreservation, Lecithin, Egg yolk, Ram semen, IVF Tris-Bazlı Sulandırıcılarda Yumurta Sarısı ve Soya Lesitininin Eritme Sonrası Koç Spermasının Kalitesi ve in vitro Fertilite Üzerine Etkisi ÖzetBu çalışmanın amacı, yumurta sarısı ve farklı lesitin konsantrasyonlarının koç spermasının dondurulabilirliği üzerine etkisini değerlendirmek ve dondurup çözdürülen koç spermasının fertilizasyon yeteneğini tespit etmektir. Kitle hareketi (3-5) ve >%70 motiliteye sahip ejakülatlar birleştirildi (pooling). Pooling yapılan sperma dört gruba bölündü ve %1, %3, %6 (L1, L3 ve L6) lesitin veya %20 yumurta sarısı (EY20) içeren sulandırıcılar ile 1/5 (sperma/sulandırıcı) oranında iki aşamalı sulandırma yöntemi kullanılarak sulandırıldı. Bu çalışmanın sonucunda; motilite ve akrozomal bozukluk oranlarının dondurma prosedüründen olumsuz yönde etkilendiği tespit edildi (P<0.001). EY20 grubunun 5°C'de ve eritme sonrası motilite değerleri L1, L3 ve L6 gruplarına göre yüksek bulundu (P<0.05). Sulandırma, soğutma, ekilibrasyon ve eritme sonrası aşamalarda motilite oranları bakımından lesitin grupları arasında farklılık saptanmadı (P>0.05). Blastosist oranları bakımından değerlendirilen in vitro fertilizasyon sonuçlarına göre, EY20 grubunun farklı lesitin gruplarından daha başarılı olduğu tespit edildi. Sonuç olarak, yumurta sarısı içeren sulandırıcı ile dondurulan koç spermasının lesitin gruplarına göre eritme sonrası spermatolojik özellikler ve embriyonik gelişim bakımından daha üstün olduğu tespit edildi.

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Section: Discussionmentioning

confidence: 99%

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Üstüner¹,

Alçay²,

Nur³

et al. 2014

Kafkas Univ Vet Fak Derg

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“…Unlike this report, our in vitro fertilization rate for C57BL/6 mice using a combination of 0.2 M raffinoseC0.1 M glycerol can be related to the better protective effect of raffinoseglycerol-skim milk combination on sperm membrane compared to 0.3 M raffinose-skim milk. Glycerol has previously been shown to be effective in protection against freezing membrane injuries in commonly used species of livestock (Parks & Graham 1992, Curry 1995 and a combination of sucrose and glycerol successfully protects embryos from various mammalian species and also bull sperm (Honadel & Killian 1988, De Leeuw et al 1993) during cryopreservation. Classic cryobiology studies suggest the use of glycerol or any permeating additive to more efficiently protect cells from freezing injuries through colligative or solution effects (Mazur 1970).…”

Section: Effects Of Cryopreservation On Sperm Qualitymentioning

confidence: 99%

Effects of cryopreservation on sperm quality, nuclear DNA integrity, in vitro fertilization, and in vitro embryo development in the mouse

Yıldız¹,

Ottaviani²,

Law³

et al. 2007

Reproduction

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Efficient freezing, archiving, and thawing of sperm are essential techniques to support large scale research programs using mouse models of human disease. The purpose of this study was to investigate the effects of variable combinations and concentrations of cryoprotectants on sperm-assessment parameters of frozen-thawed mouse sperm in order to optimize cryopreservation protocols. Sperm was frozen using combinations of 3% skim milkC0.2 or 0.3 M nonpermeating raffinose with either permeating glucose, fructose, propylene glycol, ethylene glycol, glycerol, or sodium pyruvate in CD-1, C3FeB6F1/J, B6129SF1, C57BL/6NCrIBR, 129S/SvPaslco, and DBA/2NCrIBR mice. Sperm-assessment parameters included progressive motility, plasma membrane integrity (SYBR-14CPI), in vitro fertilization rate, and in vitro embryo development rate to blastocyst. DNA content analysis of sperm was measured by the sperm chromatin structure assay (SCSA). 0.3 M raffinose with 0.1 M fructose significantly improved post-thaw sperm-assessment parameters for CD-1, C3B6F1, B6129SF1 mice (P!0.05-0.01), whereas 0.2 M raffinose with 0.1 M glycerol or 0.1 M fructose enhanced sperm assessment values for C57BL/6 and 129S mice (P!0.01), compared to 0.3 M raffinose alone. DNA fragmentation during cryopreservation was significantly increased in all strains evaluated when compared with fresh control sperm in a strain-dependent manner (P!0.01). Supplementation with permeating glycerol or fructose to the cryoprotectant (CPA) solution showed a significant protective effect to DNA integrity when cryopreserving sperm from C57BL/6 and 129S mice. Damage to sperm DNA significantly decreased the rate of in vitro embryo development to blastocyst in C57BL/6 mice. The type of monosaccharide sugar or polyols, CPA molarity, and combination of permeating and nonpermeating cryoprotectant are significant factors for improving progressive motility, plasma membrane integrity, DNA integrity, in vitro fertilization rate, and in vitro embryo development rate to blastocyst in cryopreserved mouse sperm. Reproduction (2007) 133 585-595

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“…Graham & Foote 1987, Parks & Lynch 1992, De Leeuw et al 1993, White 1993, Zeron et al 2002, Amirat et al 2005 have detailed the beneficial effects of adding lipids, cholesterol or fatty acids to biomembranes before cooling, we are unaware of any previous studies on the effect of adding CLC or other fatty acids to subzero water transport properties of cells. Although it seems reasonable to expect 'alterations' in the bovine sperm membrane transport properties in the presence of CLC, a recent study by Purdy et al (2005) found that CLC treatment did not significantly alter membrane fluidity of bovine sperm after suprazero temperature changes.…”

Section: Effect Of Extracellular Ice On Bovine Sperm Membrane Transpomentioning

confidence: 99%

Effect of glycerol and cholesterol-loaded cyclodextrin on freezing-induced water loss in bovine spermatozoa

Li¹,

Saenz

Godke

et al. 2006

Reproduction

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Recent experimental data show that incubating bovine sperm with cholesterol-loaded cyclodextrin (CLC) before cryopreservation increases the percentages of motile and viable cells recovered after freezing and thawing, compared with control sperm. In the present study, we report the effect of incubating bovine sperm with CLC on the subzero water transport response and the membrane permeability parameters (reference membrane permeability (L pg ) and activation energy (E Lp )). Water transport data during freezing of bovine sperm cell suspensions were obtained at a cooling rate of 20 8C/min under three different conditions: 1. in the absence of cryoprotective agents (CPAs); 2. in the presence of 0.7 M glycerol; and 3. in the presence of 1.5 mg/ml CLC and 0.7 M glycerol. With previously published values, the bovine sperm cell was modeled as a cylinder of length 39.8 mm and radius 0.4 mm, with osmotically inactive cell volume (V b ) of 0.61 V o , where V o is the isotonic cell volume. By fitting a model of water transport to the experimentally obtained data, the best-fit water transport parameters (L pg and E Lp ) were determined. The predicted best-fit permeability parameters ranged from L pg 5 0.02 to 0.036 mm/min-atm and E Lp 5 26.4 to 42.1 kcal/mol. These subzero water transport parameters are significantly different from the suprazero membrane permeability values (obtained in the absence of extracellular ice) reported in the literature. Calculations made of the theoretical response of bovine spermatozoa at subzero temperatures suggest that the optimal cooling rate to cryopreserve bovine spermatozoa is 45-60 8C/min, agreeing quite closely with experimentally determined rates of freezing bovine spermatozoa.

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Effects of Various Cryoprotective Agents and Membrane-Stabilizing Compounds on Bull Sperm Membrane Integrity after Cooling and Freezing

Cited by 170 publications

References 0 publications

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Effects of cryopreservation on sperm quality, nuclear DNA integrity, in vitro fertilization, and in vitro embryo development in the mouse

Effect of glycerol and cholesterol-loaded cyclodextrin on freezing-induced water loss in bovine spermatozoa

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