Cryosurvival of Human Spermatozoa Frozen in Eight Different Buffer Systems

Weidel, Louise; Prins, Gail S.

doi:10.1002/j.1939-4640.1987.tb02417.x

Cited by 68 publications

(26 citation statements)

References 15 publications

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“…Zwitterion buffers such as Tes, Hepes and Pipes have pKa values closer to the optimal freezing pH of 7.0 and vary less than Tris with temperature changes (Good ef al, 1966). Comparisons have been made between diluents based on Tris-citrate and zwitterion buffers for freezing semen of boars , bulls Parrish and Foote, 1980;Garcia and Graham, 1989a), humans (Prins and Weidel, 1986;Weidel and Prins, 1987) and turkeys , but not rams.…”

Section: Introductionmentioning

confidence: 99%

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

Molinia

Evans²,

Maxwell³

1994

Reprod. Nutr. Dev.

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Summary ― Diluents based on the zwitterion buffers Tes, Hepes and Pipes titrated to pH 7.0 with NaOH or Tris were compared with Tris-citrate diluents by assessment of post-thawing motility and acrosome integrity of frozen ram spermatozoa. Varying buffer osmolalities between 300 and 360 mosmol had no effect but there was a decrease (P < 0.001 ) in motility between 360 and 420 mosmol. There was a quadratic effect (P < 0.001 ) on motility of increasing glucose concentration in the diluent (30, 85, 140, 195 and 250 mM) with a maximum at 85 mM. At this sugar concentration, motility was higher (P < 0.001 ) for diluents containing glucose and fructose than lactose, sucrose or trehalose. Inclusion of egg yolk (optimum at 13.5% v/v; P < 0.05), centrifugation of the diluents (P < 0.01 use of low dilution rates (3 or 6-fold; P < 0.05) and freezing in pellets rather than minitubes or straws (P < 0.001 ) improved both the motility and acrosome integrity of spermatozoa in the zwitterion diluents. It is concluded that zwitterion buffers are superior to Tris-citrate in ram semen freezing diluents. zwitterion buffer / motility / acrosome integrity / frozen ram spermatozoa Résumé ― Effet de l'addition de tampons amphotères à des dilueurs sur la congélabilité des spermatozoïdes de bélier. Nous avons comparé l'efficacité de dilueurs à base de tampons amphotères TES, HEPES et PIPES ajustés à pH 7,0 avec NaoH ou tris, à celle de milieux à base de tris-citrate, en évaluant la motilité des spermatozoïdes de bélier et l'intégrité de leur acrosome après congélation. Entre 300 et 360 milliosmoles, l'osmolarité des tampons n'a aucun effet. Au-delà, et jusqu'à 420 milliosmoles, on observe une baisse significative (P < 0, 001) de la motilité. On obtient un effet biphasique sur la motilité en augmentant la concentration de glucose dans le dilueur (30, 85, 140, 195, 250 mM) avec un maximum pour 85 mM. À cette concentration, la motilité est plus élevée avec le milieu contenant du glucose et du fructose qu'avec ceux contenant du lactose, du sucrose ou du tréhalose (P < 0,001). L'addition de jaune d'œuf (optimum à 13,5% vlv; P < 0,05), la centrifugation des dilueurs (P < 0,01), l'utilisation d'un taux de dilution peu élevé (3 ou 6 fois ; P < 0,05) et la congélation en pastilles plutôt qu'en minitubes ou en minipaillettes (P < 0,001) améliorent la motilité et l'intégrité de l'acrosome des spermatozoïdes congelés dans des milieux contenant un tampon amphotère. Cette étude montre donc que les tampons amphotères sont plus efficaces que le tampon tris-citrate pour les dilueurs utilisés pour la congélation du sperme de bélier.bélier / mobilité l acrosome / sperme congelé / dilueur / tampon * Current address:

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Section: Introductionmentioning

confidence: 99%

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

Molinia

Evans²,

Maxwell³

1994

Reprod. Nutr. Dev.

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“…A variety of cryoprotective media have been designed in an attempt to overcome the cellular damage caused by cryopreservation [29,30]. LDL is added to the freezing extender instead of egg yolk because it better preserves the characteristics of sperm in bulls [16,17] and boars [18,19].…”

Section: Discussionmentioning

confidence: 99%

Characteristics of Okinawan Native Agu Pig Spermatozoa After Addition of Low-Density Lipoprotein to Freezing Extender

Yamauchi

Lay

Azuma

et al. 2009

Journal of Reproduction and Development

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Abstract. Technical refinement of boar sperm cryopreservation is indispensable for effective breeding of the rare Okinawan native pig, the Agu. The objective of the present study was to determine whether addition of low-density lipoprotein (LDL) extracted from hen egg yolk to the freezing extender improves the characteristics of cryopreserved Agu spermatozoa. Ejaculated Agu sperm frozen in extender supplemented with 2, 4, 6, 8 or 10% LDL instead of egg yolk was thawed, and the post-thaw sperm characteristics were evaluated. Treatment with 4-8% LDL during cooling and freezing significantly increased the intracellular cholesterol content, as compared to that of sperm frozen in extender containing 20% egg yolk (P<0.05). Higher potential resistance to cell damage from cryoinjury was also observed in sperm frozen in extender supplemented with LDL: the integrities of plasmalemma and DNA, mitochondrial activity and proteolytic activity of the acrosomal content in the post-thaw sperm were superior to those of sperm that were not treated with LDL. Moreover, the percentages of total motile sperm and the extent of rapid progressive motility at 1 and 3 h after incubation were markedly higher in sperm treated with 4 or 6% LDL, and these sperm also had more ATP. However, LDL did not inhibit in vitro sperm penetrability, even though the cholesterol content of post-thaw sperm was higher after treatment with LDL. These findings indicate that addition of 4-6% LDL instead of egg yolk to the freezing extender improves the post-thaw characteristics of Agu sperm by protecting sperm against cold shock damage during cryopreservation. Key words: Boar spermatozoa, Cell damage, Cryopreservation, Low-density lipoproteins (LDL) (J. Reprod. Dev. 55: [558][559][560][561][562][563][564][565] 2009) ecently, the male reproductive ability (i.e., semen volume, sperm quality and sperm concentration) of the Okinawan native pig, the Agu, has markedly decreased because of repeated inbreeding within a minority of the closed population for the past 20 years, which has resulted in a marked reduction in reproductive efficiency under natural mating conditions. Consequently, longterm preservation and storage of Agu sperm for artificial insemination (AI) has become a subject of interest because it permits insemination of a large number of females in a short period of time, avoids transmission of disease and conserves valuable germplasm stocks. However, the poor post-thaw survival of boar sperm has limited the success of AI with frozen sperm and has prevented its widespread acceptance in commercial breeding [1,2].Boar sperm are extremely sensitive to the damaging effects of cold shock [3]. The susceptibility of sperm to cryopreservation differs across species: rooster, human, monkey, rabbit and dog sperm are more resistant to cold shock than bull, ram, horse and boar sperm [3][4][5][6]. According to previous reports [4,6], the ratio of proteins to phospholipids in the plasma membrane is lowest in rooster sperm, at 0.46, moderate in bull and stallion sperm ...

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“…Tris:Citrate-This buffer combination has been used to freeze semen from a variety of domestic species including ram [44,107], boar [41], bull [43] and turkey [42], though post-thaw motility and acrosome integrity was often lower than that obtained from other zwitterionic buffers studied, like HEPES, TES and PIPES [44,107]. The buffer was also used to successfully freeze human spermatozoa [108,109]. In early attempts to achieve fertilization in vitro in hamster, Bavister examined the use Tris-Citrate buffer (25 mM).…”

Section: Buffers In Ivfmentioning

confidence: 99%

Biological pH buffers in IVF: help or hindrance to success

Will

Clark

Swain

2011

J Assist Reprod Genet

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Purpose Minimizing environmental stress helps maintain cellular homeostasis and is a crucial component in optimizing embryo development in vitro and resulting ART success. One stressor of particular interest is pH. Biologic buffers, such as HEPES and MOPS, are valuable tools for stabilizing pH. The objective of this manuscript is to summarize efficacy and impact of various pH buffers used during IVF lab procedures Methods Keyword searches were performed using Pubmed and Medline and relevant literature reviewed. Results Various pH buffers have been used with varying degrees of success for gamete and embryo processing in a variety of animal species, as well as in human. Conclusion Though biologic buffers off a means to improve pH stability, not all buffers may be appropriate for use with gametes and embryos. Specific buffers may have undesired effects, and these may be buffer, species, cell type or concentration dependent. Continued research is needed to further refine and improve the use of biologic buffers for use in human ART.

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Cryosurvival of Human Spermatozoa Frozen in Eight Different Buffer Systems

Cited by 68 publications

References 15 publications

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

Characteristics of Okinawan Native Agu Pig Spermatozoa After Addition of Low-Density Lipoprotein to Freezing Extender

Biological pH buffers in IVF: help or hindrance to success

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