• Title, Summary, Keyword: cryoprotectant

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The Effect of Dimethyl-Sulfoxide and Sucrose as a Cryoprotectant on the Adenosine Triphosphate and Ultrastructure of Bovine Oocytes Matured In Vitro

  • Tsuzuki, Yasuhiro;Hisanaga, Mio;Ashizawa, Koji;Fujihara, Noboru
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.10
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    • pp.1353-1359
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    • 2001
  • The present study was undertaken to assess the influence of dimethyl-sulfoxide plus sucrose solution as a cryoprotectant on the adenosine triphosphate (ATP) content, the ultrastructure and the embryonic development of bovine oocytes matured in vitro. We measured the amount of ATP in cumulus cells enclosed oocytes (CO) or denuded oocytes (DO) equilibrated with or removed from the cryoprotectant (1.5 M DMSO + 0.25 M sucrose + 20% fetal bovine serum in physiological saline). As a result, the ATP contents in both CO and DO, equilibrated with the cryoprotectant, were significantly lower (p<0.05) than that of the each control group. However, ATP content of DO was recovered to the level of the control group ailer removal of the cryoprotectant, but failed to restore for CO. In the observation of the ultrastructure by a transmission electron microscope, all of the mitochondria in the ooplasm of CO and DO equilibrated with the cryoprotectant were swollen with peripherally located cristae following decreased electron density. However, a large proportion of these swollen mitochondria were restored to the normal shape which can be observed usually in the control group after removal from the cryoprotectant. To the contrary, the morphology of many mitochondria of the cumulus cells in CO were not recovered to that of the control group after removal of the cryoprotectant. CO with removed cryoprotectant had significantly lower embryonic development up to the blastocysts stage (p<0.05) after in vitro fertilization compared with that in the control group. These results suggest that the addition and removal of a cryoprotectant has a negative effect for the ATP content of cumulus enclosed oocytes. One of the factor(s) causing the lower embryonic development after removal of cryoprotectant, may be associated with ATP metabolism.

Effect of Substitution of Chicken Breast for Alaska Pollack on Physico-chemical Characteristics and Quality in Surimi-like Materials Contained Different Cryoprotectants

  • Jin, Sang-Keun;Choi, Young-Jun;Hur, Sun-Jin
    • Food Science of Animal Resources
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    • v.31 no.2
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    • pp.215-223
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    • 2011
  • Surimi-like samples were divided into four groups (C, surimi-like material made from Alaska Pollack with all cryoprotectant ingredients; T1, surimi-like material made from chicken breast with sugar and a sorbitol-free cryoprotectant; T2, surimi-like material made from chicken breast with a sugar-free cryoprotectant; T3, surimi-like material made from chicken breast with all cryoprotectant ingredients). Water and protein content were lower in Alaska Pollack surimi-like material (C) than those in chicken breast surimi-like material. Centrifuge loss and cooking loss were higher in C than those in chicken breast surimi-like material. Lipid oxidation (thiobarbituric acid reactive substances) was lower in T3 than others during storage. In a sensory evaluation, overall acceptability was significantly higher in C than those in other samples during storage. As a result, we found that the raw material composition (Alaska Pollack or chicken breast) had a large influence on the physico-chemical characteristics and quality of surimi-like materials, whereas cryoprotectant composition may have less influence on the physico-chemical characteristics and quality of surimi-like materials.

The Effect of Cryoprotectants on the Properties of Pacific Sand Lance Ammodytes personatus Girard Surimi During Frozen Storage

  • Yoo, Byung-Jin
    • Fisheries and aquatic sciences
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    • v.17 no.3
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    • pp.291-298
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    • 2014
  • We investigate the effects of cryoprotectant mixtures on the quality of sand lance surimi (SLS) during storage at $-30^{\circ}C$. We monitored freeze-induced denaturation of myofibrillar protein in SLS and examined the texture profile of SLS gel. Freeze-induced denaturation was assessed by evaluating SLS $Ca^{+2}$-ATPase activity. SLS gels prepared with sorbitol or sucrose and a mixture of both as cryoprotectant. Higher concentrations of cryoprotectants resulted in significantly higher residual SLS $Ca^{+2}$-ATPase activity at the same storage time (P < 0.05). Residual $Ca^{2+}$-ATPase activity of SLS prepared with sorbitol was higher than that of sucrose when cryoprotectant concentration and storage period were same. A blend of sorbitol and sucrose resulted in a stronger cryoprptective effect of SLS myofibrillar protein than did sorbitol or sucrose alone. The presence of a phosphate compound in SOP (3% sorbitol + 0.2% phosphate compound) resulted in higher SLS $Ca^{2+}$-ATPase activity than that of did 5% sorbitol. The hardness, brittleness, and elasticity values and a folding test of the SLS gels were significantly affected by cryoprotectant concentrations and the storage time. Preference scores and acceptance for texture in a sensory evaluation of the SLS gels increased with increasing sorbitol or sucrose concentration.

Survival Rates of Trochophores from Pearl Oyster, Pinctada fucata martensii and Pacific Oyster, Crassostrea gigas Immersed in Pour Kinds of Cryoprotectant (4종류의 동해 방지제에 침지 한 진주조개, Pinctada fucata martensii와 참굴, Crassostrea gigas 담륜자의 생존율)

  • CHOI Youn Hee;CHANG Young Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.476-480
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    • 1999
  • Experiments were carried out to evaluate the tolerance of trochophores for pearl oyster, Pinctada fucata martensii and Pacific oyster, Crassostrea gigas using different concentrations of cryoprotectants : dimethyl sulfoxide (DMSO), ethylene glycol, glycerol and 1,2-propanediol. Each cryoprotectant with different concentrations was exposed for 5, 10, 15 and 20 minutes of immersion time. Survival rates were increased with decreased concentrations of cryoprotectant and decreased immersion time, and these differed from types of cryoprotectant. Survival rates of Pacific oyster trochophores were higher in DMSO and ethylene glycol, while those of pearl oyster trochophores were higher in glycerol and 1,2-propanediol. In case of trochophores from Pacific oyster, when 0.2 M sucrose was added in each cryoprotectant the survival rates were increased significantly.

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The Effects of Cryoprotectants on Motility and Viability Kinetics of Liquid Boar Semen at $4^{\circ}C$ (액상 보존액 내 동결 보호제가 $4^{\circ}C$에 보관된 액상 돼지 정액의 운동성과 생존성에 미치는 효과)

  • Jeon, Yu-Byeol;Park, Kwang-Woo;Kwak, Seong-Sung;Jeong, Seung-A;Yoon, Jun-Chul;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.27 no.1
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    • pp.51-55
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    • 2012
  • The objective of this study was to investigate the motility and kinematics of boar sperm that while stored at 4C. The samples of fresh boar semen were place into an extender, Androhep, and stored at $4^{\circ}C$. In three of these samples, cryoprotectants were added. The sperm's motilities and kinematics were evaluated by using microscope (${\times}400$) and the viability status was evaluated by using with eosin staining method. The 5 sample groups are; Goup A:Androhep (extender), stored at $17^{\circ}C$. Group B:Androhep (extender), stored at $4^{\circ}C$. Group C:Androhep (extender), + 3% glycerol (cryoprotectant), stored at $4^{\circ}C$. Group D:Androhep (extender), + 3% DMSO (cryoprotectant), stored at $4^{\circ}C$. Group E:Androhep (extender), + 3% ethylene glycol (cryoprotectant), stored at $4^{\circ}C$. In group A, the sperm's motility was reduced. On day one the sperm's motility was ($85.7{\pm}2.3$) and day 5 the motility was ($43.9{\pm}3.3$). In group B, C and D the sperm's motility were reduced to 0 on day 5. In group E the sperm's percentage of motility decreased. On day one the sperm's motility was ($42.0{\pm}0.5$) and day 5 the motility was ($2.3{\pm}0.3$). When comparing cryoprotectant in samples of boar sperm there is a slight improvement in the results when the use of Androhep Lite (extender), + 3% ethylene glycol (cryoprotectant), stored at $4^{\circ}C$ are used. Based on these results, ethylene glycol can protect sperm from heat shock at $4^{\circ}C$, but not satisfactory level. However, it showed the possibilities of liquid semen preservation at $4^{\circ}C$ by using cryoprotectant.

Development of Cell Line Preservation Method for Research and Industry Producing Useful Metabolites by Plant Cell Culture

  • Cho, Ji-Suk;Chun, Su-Hwan;Lee, Song-Jae;Kim, Ik-Hwan;Kim, Dong-Il
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.5
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    • pp.372-378
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    • 2000
  • The cell culture of Angelica gigas Nakai producing decursin derivatives and immunostimulating polysaccharides was preserved in liquid nitrogen after pre-freezing in a deep freezer at -70$^{\circ}C$ for 480 min. The effects of the cryoprotectant and pretreatment before cooling were investigated to obtain the optimal procedure for cyropreservation. When compared to mannitol, sorbitol, or NaCl with a similar osmotic pressure, 0.7 M sucrose was found to be the best osmoticum for the cryopreservation of A. gigas cells. In the pre-culture medium, the cells in the exponential growth phase showed phase showed the best post-freezing survival after cryopreservation. A mixture of sucrose, glycerol, and DMSO was found to be an effective cryoprotectant and a higher concentration of the cryoprotectant provided better cell viability. When compared with the vitrification, the optimum cryopreservation method proposed in this study would seem to be more effective for the long-term storage of suspension cells. The highest relative cell viability established with the procedure was 89%.

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Selection of Cryoprotectant for the Cryopreservation of Trochophores and Early D-shaped Larvae of Surf Clam, Spisula sachalinensis (북방대합, Spisula sachalinensis 담륜자와 초기 D상 유생의 냉동보존을 위한 동해방지제의 선택)

  • KIM Young Sin;CHOI Youn Hee;LEE Jeong Yong;CHANG Young Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.597-599
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    • 2000
  • This study was performed to find out the cryoprotectants for cryopreservation of trochophores and early D-shaped larvae of surf clam, Spisula sachalinensis. Dimethyl sulfoxide (DMSO), ethylene glycol, 1,2-propanediol and methanol were used as cryoprotectant Each cryoprotectant was made to 1.0 M, 2.0 M, 3.0 M with dilution of 0.2 M fructose and 0.2 M sucrose. The trochophoers and early D-shaped larvae were immersed in each preparation for 10 minutes to reach equilibration and cryopreserved in liquid nitrogen. Survival rates of post-thawed trochophores and early D-shaped larvae in 2.0 M DMSO with 0.2 M sucrose were the highest as $91.4{\%}\;and\;78.9{\%}$, respectively.

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Freezing Preservation of Liquid Egg by Freezing Point Depression (빙점강하에 의한 액란의 냉동저장에 관한 연구)

  • Lee, Young-Chun;Lee, Kyung-Hae
    • Korean Journal of Food Science and Technology
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    • v.20 no.4
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    • pp.594-599
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    • 1988
  • Methods by which liquid egg could be stored in liquid state at frozen storage temperature$(-15^{\circ}C)$ with selected cryoprotectants and enzyme treatment were investiated, and quality changes in samples during storage were examined. The concentration of cryoprotectants (45% fructose and 55% glucose) to be added to egg yolk and whole egg to store them at $-15^{\circ}C$ in unfrozen state were 45.2% and 70.3%, respectively. Changes in consistency, precipitation of protein and microstructure of egg samples during storage indicated that adding cryoprotectants to liquid egg could effectively inhibit development of gelation during storage at $-15^{\circ}C$. Treating liquid egg with 0.15% papain could inhibit gelation during storage to some extent.

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Diluents and Cryoprotectants for Cryopreservation of Filefish Thamnaconus modestus Sperm

  • Le, Minh-Hoang;Lim, Han-Kyu;Min, Byung-Hwa;Son, Maeng-Hyun;Lee, Jung-Uie;Chang, Young-Jin
    • Journal of Aquaculture
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    • v.21 no.1
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    • pp.54-59
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    • 2008
  • The present study aimed to find the best diluent and cryoprotectant for sperm cryopreservation of filefish Thamnaconus modestus. Two kinds of artificial seminal plasma(ASP1, ASP2), 0.3 M glucose and marine fish Ringer's solution(MFRS) were employed as diluent. Dimethyl sulfoxide(DMSO) and methanol as cryoprotectant were selected for sperm cryopreservation. Sperm was diluted at the ratio of 1:3 with diluents containing cryoprotectants and adjusted for final concentration at 5%, 10%, 15% and 20%. Mixed milt was frozen at liquid nitrogen vapor after equilibration for 5 min. The highest motility($40.5{\pm}2.8%$) and swimming speed($81.5{\pm}4.1{\mu}m/s$) of frozen/thawed sperm were observed in ASP1 diluent containing 10% DMSO and in ASP2 containing 15% DMSO, respectively. Results showed that cryopreservation with ASP as diluent and DMSO as cryoprotectant could be adopted for long term storage of filefish sperm.

Screening of Lactic Acid Bacteria for the Development of Probiotics and the Effect of Cryoprotectant Agents (생균제 개발을 위한 유산균의 선별 및 동결건조 보호제의 효과)

  • 임유범;백남수;김영만
    • The Korean Journal of Food And Nutrition
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    • v.14 no.5
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    • pp.441-445
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    • 2001
  • For the developement of probiotics, three strains haying psychorophilic and salt tolerant characteristics were isolated from Kimchi. Among the isolated strains, MGl9, MG89 and MG208 were identified as Lactobacillus brevis, Enterococcus faecium and Lactobacillus plantarum, respectively. The relationship between cryoprotectant and the viability of freeze dried strains has been studied. The most effective cryoprotectant for MGl9 was found to be 10% skim milk contained 1% soluble starch haying the survival rate of 71.4%. In MG89, 10% skim milk contained 1% sucrose and 10% skim milk contained l% fructose in MG208 were showed effective cryoprotectant having the survival rate 68.9% and 64.7%. respectively These results suggest that these isolated strains can play an important role as probiotics in aquaculture.

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