• Asian-Australasian Journal of Animal Sciences
• /
• v.14 no.10
• /
• pp.1353-1359
• /
• 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.

• Food Science of Animal Resources
• /
• v.31 no.2
• /
• pp.215-223
• /
• 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.

• Fisheries and Aquatic Sciences
• /
• v.17 no.3
• /
• pp.291-298
• /
• 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.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.32 no.4
• /
• pp.476-480
• /
• 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.

• Journal of Embryo Transfer
• /
• v.27 no.1
• /
• pp.51-55
• /
• 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.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.33 no.6
• /
• pp.597-599
• /
• 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.

• Korean Journal of Food Science and Technology
• /
• v.20 no.4
• /
• pp.594-599
• /
• 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.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.5 no.5
• /
• pp.372-378
• /
• 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%.

• Journal of Aquaculture
• /
• v.21 no.1
• /
• pp.54-59
• /
• 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.

• Clinical and Experimental Reproductive Medicine
• /
• v.25 no.3
• /
• pp.261-268
• /
• 1998

The present study was to assess the effect of ultrarapid freezing on the development of 1-cell mouse zygote using cryoprotectants, DMSO (dimethyl sulfoxide) or PROH (1,2-propanediol). We investigated the effect of the type and concentration of cryoprotectant, and of the temperature and time of prefreezing equilibration on their capacity to develop to the blastocyst stage in vitro. The concenration, the equilibration temperature, and the exposure time seemed to serve as an important factor in ultrarapid freezing of 1-cell mouse zygotes. In addition to the exposure time and the concentration of cryoprotectant appeared to playa key role in the development of the embryo. In general, the development of the embryo was more effective at $3^{\circ}C$ than $23^{\circ}C$ and 4.5 M than 3 M for 3 to 5 minutes. At $23^{\circ}C$ the development of the embryo was stimulated by DMSO while at $3^{\circ}C$ it was stimulated by PROH. Thus it has been suggested that there exists a correlation between the concentration of cryoprotectants and exposure time in the development of the embryo. In conclusion, we found that for ultrarapid freezing of mouse 1-cell embryos in DMSO, or PROH-based solution, viability shown optimum depending on the cryoprotectant, the concentration of the cryoprotectant and on the temperature and the duration of equilibration.