Effect of Sucrose, Trehalose, Hypotaurine, Taurine, and Blood Serum on Survival of Frozen Bull Sperm

Chen, Y.; Foote, R.H.; Brockett, C.C.

doi:10.1006/cryo.1993.1042

Cited by 143 publications

(78 citation statements)

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“…Therefore, the addition of a cryoprotectant to the cells is important. Among the cryoprotectants used, trehalose, which is a sugar that is utilized in cell cryopreservation, is significant (10)(11)(12)(13)(14)(15). Trehalose is utilized in many fields, such as in the pharmaceutical industry (contained in various commercially available therapeutic products), the food industry (in confectionary products) and in the cosmetics industry (for example, in bath oils, creams and lotions) (23).…”

Section: Discussionmentioning

confidence: 99%

Effect of trehalose on cryopreservation of pure peripheral blood stem cells

Martinetti¹,

Colarossi²,

Buccheri³

et al. 2017

Biomedical Reports

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Abstract. Stem cells are an important tool for the study of hematopoiesis. Despite developments in cryopreservation, post-thaw cell death remains a considerable problem. Cryopreservation protocol should limit cell damage due to freezing and ensure the recovery of the functional cell characteristics after thawing. Thus, the use of cryoprotectants is essential. In particular, the efficacy of trehalose has been reported for clinical purposes in blood stem cells. The aim of the current study was to establish an efficient method for biological research based on the use of trehalose, to cryopreserve pure peripheral blood stem cells. The efficacy of trehalose was assessed in vitro and the cell viability was evaluated. The data indicate that trehalose improves cell survival after thawing compared with the standard freezing procedure. These findings could suggest the potential for future trehalose application for research purposes in cell cryopreservation. IntroductionStem cells in biological research provide a significant source of information and, thus, contribute to the development of novel therapeutic strategies. In particular, the ability to cultivate stem cells and human hematopoietic progenitor cells in vitro is the fundamental basis for investigating hematopoiesis. An improved understanding of the mechanisms that regulate cell proliferation and differentiation during the stages of hematopoiesis would further elucidate the molecular characteristics of diseases (which are characterized by excessive expansion or a functional defect of certain immature blood components), and facilitate the identification of substances that are able to specifically protect healthy cells from the action of cytotoxic drugs.Hematopoietic stem and progenitor cells (HSPCs) may be isolated from peripheral blood (PB), bone marrow (BM) or umbilical cord blood (CB) (1).Although significant improvements to cell cryopreservation procedures have been achieved (2), the improvement of current cryopreservation protocols that lead to a high mortality rate after thawing for the crystal formations that arise during freezing is a primary goal. Specifically, fast cooling forms intracellular ice crystals, which results in cell destruction and slow cooling forms ice crystals in the extracellular space, with consequent cellular dehydration. Selection of a cryoprotectant, as well as a suitable freezing rate serves to protect cells from these adverse effects (3,4).Cryoprotectants are divided into two classes: Penetrating and nonpenetrating (5). The penetrating cryoprotectants include glycerol and 1,2-propanediol and dimethyl sulfoxide (Me 2 SO); the latter is commonly used for HSPC cryopreservation (6).The non penetrating cryoprotectants comprise polyvinyl pyrrolidone, trehalose, fructose, sucrose and glucose.Trehalose is a non-toxic disaccharide of glucose that preserves the structural integrity of the cells during freezing and thawing (7). Specifically, trehalose is found in numerous organisms, such as nematodes and yeasts, which are capable of surviving du...

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Section: Discussionmentioning

confidence: 99%

Effect of trehalose on cryopreservation of pure peripheral blood stem cells

Martinetti¹,

Colarossi²,

Buccheri³

et al. 2017

Biomedical Reports

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“…Also Dolezalova et al (2015) in previous study confirmed these results. However, Chen et al (1993) obtained a successful protocol for freezing bull semen commercially cooling sperm by the rate of 15°C/min from +5 to -100°C, followed by transfer to liquid nitrogen. In general, semen cryopreservation protocols use freezing curves ranging from 10 to 100°C/min, resulting in good survival rates after thawing.…”

Section: Discussionmentioning

confidence: 99%

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Doležalová¹,

Stádník²,

Biniová³

et al. 2016

Czech J. Anim. Sci.

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ABSTRACT:The objective was to determine effects of equilibration length and freezing curve type as well as their interactions on motility and live spermatozoa proportion in bull sperm after thawing. The ejaculates of 6 sires were repeatedly collected. Fresh semen was diluted with one extender and divided into 3 groups equilibrated for 30, 120, and 240 min. Subsequently, half straws of each group were frozen using standard 3-phase or 2-phase freezing curve differing in the rate of temperature decrease. Th e spermatozoa motility (M) was evaluated immediately after thawing and at 30, 60, 90, and 120 min of thermodynamic test (TDT). Live spermatozoa proportion was evaluated after thawing and at the end of TDT. Average of spermatozoa motility (AM), decrease of spermatozoa motility (MD), average proportion of live spermatozoa (ALS), and decrease of live spermatozoa proportion (DLS) through the TDT were calculated. Significant inter-sire differences in AM (0.45-17.0%; P < 0.05-0.01), MD (0.76-12.57%; P < 0.05-0.01), and ALS (0.99-23.8%; P < 0.01) were detected. The longest equilibration ensured the highest M during TDT and AM (+2.72 and +4.58%; P < 0.05-0.01), however higher MD (+4.06%; P < 0.01) compared to standard length as well. Straws freezed using 2-phase curve achieved higher M through TDT, AM (+7.3%; P < 0.01) as well as ALS (+11.77%; P < 0.01). The 2-phase curve presented higher M compared to the 3-phase freezing curve within all equilibration lengths. Significant differences in AM, MD,; P < 0.05-0.01) between equilibration length vs freezing curve interactions were determined. Results document the importance of equilibration length, freezing curve, and their interaction effect on live spermatozoa proportion and sperm motility after thawing as well as necessity of individual conditions for bulls semen processing and insemination doses production.

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“…The beneficial effects of the trehalose on the postthawing viability of mammalian sperm cells have been reported in many studies (6). In some reports, trehalose conferred better cryoprotection and post-thawing fertilizing ability of ram, bull and mouse sperm due to diminished death and damage of sperm (9). Molinia et al (20) demonstrated that the motility of frozen-thawed ram spermatozoa was higher in the presence of trehalose than that of glucose when glycerol is not incorporated in the diluent.…”

Section: Discussionmentioning

confidence: 99%

The role of different trehalose concentrations and cooling rates in freezing of ram semen

Ongun;BUCAK

2009

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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Summary:The aim of this study was to determine the influence of different cooling rates and trehalose concentrations on standard semen parameters after the freezing-thawing of ram spermatozoa. Ejaculates collected using artificial vagina from 4 Akkaraman rams were evaluated and pooled at room temperature. Semen samples diluted with a Tris-based extender containing different concentrations (50, 100 and 150 mM) of trehalose and no additives (control) were cooled to 5°C and frozen at slow (a rate of 0.5°C/min from 5°C to -25°C; -50°C/min from -25°C to -130°C), and fast (a rate of 10°C/min from 5°C to -25°C; -50°C/min from -25°C to -130°C) cooling rates and plunged into liquid nitrogen. Frozen straws were thawed individually at 37°C for 20 sec in a water bath for evaluation. In conclusion, 50 and 100 mM trehalose and slow cooling rate were found to significantly (p<0.05) improve the freezability of ram spermatozoa which were assessed regarding post-thawing sperm motility, morphologic sperm abnormalities, viability and membrane integrity. But, 100 mM trehalose concentration and slow cooling rate were observed superior than other treatment groups.Key words: Fast freezing, ram semen, semen parameters, slow freezing, trehalose. Koç spermalarının dondurulmasında farklı trehaloz konsantrasyonu ve soğutma oranlarının rolüÖzet: Bu çalışmanın amacı dondurma/çözdürme sonrası farklı soğutma oranları ve trehaloz konsantrasyonlarının standart spermatolojik parametreler üzerine etkisini saptamaktır. Sun'i vagina kullanılarak 4 Akkaraman koçtan toplanan ejakülatlar oda sıcaklığında değerlendirildi ve karıştırıldı. Farklı konsantrasyonlarda (50, 100 ve 150 mM) trehaloz içeren ve içermeyen (control) tris bazlı solusyonla sulandırılmış sperma örnekleri 5°C'a soğutuldu ve yavaş (dakikada 0.5°C ile 5°C'dan -25°C'a; dakikada -50°C ile -25°C'dan -130°C'a) ve hızlı (dakikada 10°C ile 5°C'dan -25°C'a; dakikada -50°C ile -25°C'dan -130°C'a) soğutma oranlarında donduruldu ve sıvı azota daldırıldı. Dondurulmuş payetler değrlendirme için bireysel olarak su banyosunda 37°C'da 20 saniyede çözdürüldü. Sonuç olarak, çözdürme sonrası spermatozoa motilitesi, morfolojik spermatozoa anomalileri, canlılığı ve membrane bütünlüğü değerlendirilen koç spermatozoasının dondurulabilirliğini, 50 ve 100 mM trehaloz ile yavaş dondurma oranının önemli ölçüde (p<0.05) geliştirdiği bulundu. Fakat, 100 mM trehaloz konsantrasyonu ve yavaş soğutma oranının diğer tedavi gruplarından daha üstün olduğu gözlendi. Anahtar sözcükler: Hızlı dondurma, koç sperması, sperma parametreleri, yavaş dondurma, trehaloz..

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Effect of Sucrose, Trehalose, Hypotaurine, Taurine, and Blood Serum on Survival of Frozen Bull Sperm

Cited by 143 publications

References 0 publications

Effect of trehalose on cryopreservation of pure peripheral blood stem cells

Effect of trehalose on cryopreservation of pure peripheral blood stem cells

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

The role of different trehalose concentrations and cooling rates in freezing of ram semen

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