• Title/Summary/Keyword: Cryoprotectants

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Effect of Cryoprotectants on the Quality Characteristics of Chicken Breast Surimi Manufactured by pH Adjustment during Freezing Storage (pH 조절법으로 제조한 닭가슴살 수리미의 저장 중 품질특성에 미치는 냉동변성방지제 첨가 효과)

  • Jin, Sang-Keun;Kim, Il-Suk;Choi, Yeung-Joon;Yang, Han-Sul;Park, Gu-Boo
    • Food Science of Animal Resources
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    • v.27 no.3
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    • pp.320-328
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    • 2007
  • This study was conducted to determine the effect of pH adjustment and the addition of cryoprotectants on the quality characteristics of chicken breast surimi. We prepared surimi from Alaska pollack, as a the control, by two time washing times and the addition of cryoprotectants. Different preparations of surimi were manufactured by adjusting to pH 11.0 and the addition of different addition cryoprotectants during frozen storage (T1 : 5% sorbitol and 0.3% polyphosphate, T2: 4% sugar, 5% sorbitol and 0.3% polyphosphate, and T3: 2% salt, 4% sugar, 5% sorbitol and 0.3% polyphosphate). The moisture content was significantly lower in the control and T3 samples. The crude protein content was increased with storage times. The crude protein was higher in the control. The water-holding capacity, myofibrillar protein and shear force were significantly higher in T3 than other surimi samples. All gel characteristics were significantly higher in the control and T3 than other surimi samples. pH 11.0 adjusted chicken breast surimi had greater lightness than the control, and T3 samples had the highest lightness and whiteness. Sensory evaluations were significantly higher in the control and T3 than the other samples. The gel, and physical characteristics and sensory evaluation of T3 were similar to the control. T3 samples had superior color and pH than the control Alaska pollack surimi.

Studies on the Survival and In Vitro Fertilization Rates of Frozne-Thawed Porcine Embryos and Immature Oocytes (돼지 수정란 및 미성숙 난자의 동결융해 후의 생존율에 관한 연구)

  • 김상근;이명헌;남윤이
    • Korean Journal of Animal Reproduction
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    • v.22 no.2
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    • pp.187-194
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    • 1998
  • In order to determine suitable conditions for rapid freezing of porcine embryos, the kind and concentration of cryoprotectants, sucrose concentrations, equilibration time and thawing temperature in freezing medium were examined in relation to the survival of frozen-thawed oocyte and embryos. The results obtained are as follows : 1. The suitable concentrations of cryoprotoctant in the freezing medium which consisted of TCM-199+20% FCS were 1.5M for glycerol, 2.0M for DMSO, 2.5M for ethylene glycol, and 2.0M for propanediol. The sucrose concentration of 0.25M in the medium was found to optimal because the survival rate was markedly higher at this concentration when compared to the others. The survival rate was relatively high when the frozen embryos were thawed at 30$^{\circ}C$ in the freezing medium containing 2.5M cryoprotectants. The equilibration periods of 2.0 and 5.0 minutes revealed the higher survival in the media containing 1.5 or 2.1M glycerol when compared to 10 and 15 minutes. 2. The fertilization rates of frozen-thawed follicular oocytes which matured in vitro for 1, 12, 24 and 48 hours were 6.7~26.7% depending on the maturation time, and the rates were relatively high for those matured for a short period of time. The survival rates of frozen-thawed oocytes which matured in vitro for certain periods and fertilized were 10.0~30.0% depending on the maturation time.

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Preservation of Marine Heterotrophic Bacteria by Using a Deep-freezing Method

  • Park, Shin-Hye;Lee, Hyun-Sang;Lee, Hong-Kum
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.240-243
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    • 2001
  • The effect of cryoprotectants and suspending solutions on the preservation of marine heterotophic bacteria was investigated. Six halotolerant and four halophilic bacterial isolates suspended in either distilled water or artificial seawater were preserved in glycerol and dimethylsulfoxide at -70$\^{C}$, respectively. After one year of preservation, the recovery rates on the appropriate agar plates were estimated. The survival rate was found to be dependent on the strain tested, regardless of the preservation conditions tested.

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Studies on the In viiro Culture and Freezing of Mouse Embryos (마우스 수정란의 체외배양 및 동결보존에 관한 연구)

  • Lee Bong-Soo;Chang kyung-Jin
    • Journal of the korean veterinary medical association
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    • v.27 no.3
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    • pp.169-175
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    • 1991
  • This study was performed to investigate the viability of 4-cell, 8-cell and morula embryos after In vitro Culture and to compare the cryoprotective effectiveness of various cryoprotectants such as glycerol, DMSO and ethylene glycol in mouse. The results w

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Studies on the Survival and In Vitro Fertilization Ratesof Frozen-Thawed Bovine Immature Oocytes (소 동결 미성숙 난자의 생존율과 체외수정율에 관한 연구)

  • 김상근;유상식;김용섭
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.207-211
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    • 1998
  • 본 연구는 소 미성숙 난자의 동결에 있어서 미성숙 난자의 1, 2, 4, 8 시간 성숙배양한 다음 동결 융해 후 체외배양하였을 때 체외수정율과 생존율을 조사하고저 수행하였다. 미성숙 난자들 1∼8시간 성숙배양후 동결융해 및 체외수정시켰을 때 수정율은 12.8∼30.9%로서 단시간의 체외성숙 배양이 높게 나타났으며(23.8∼28.8%), 또한 체외배양시 생존율은 11.8∼29.9%를 나타냈다.

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Cryopreservation of Porcine Embryos using Glycerol, Egg Yolk and Trehalose (Glycerol, 난황 및 Trehalose를 이용한 돼지유정란의 동결)

  • 장원경;박수봉;이명식;김태현;박용윤;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.43-48
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    • 1995
  • The purpose of this study was to examine the suvival rates of cryopreserved porcine embryos collected on Day 6 (Day 0=onset of estrus) with various cryprotectants. Eighty two embryos at different stages, ranged from expanded blastocyst to hatched blastocyst, were allocated to 6 experimental groups in different combinations of cryoprotectants glycerol, egg yolk and/or trehalose. Porcine embryos were cryopreserved using conventinal slow freezing precedures. The embryos were equilibrated with one of the freezing solutions, cooled from 25 to -7$^{\circ}C$ at 1$^{\circ}C$/min, seeded at -7$^{\circ}C$ frozen to -36$^{\circ}C$ at 0.5$^{\circ}C$/min, and then plunged into liquid nitrogen. The frozen embryos were thawed by immersion in 37$^{\circ}C$ water and the cryoprotectants were removed by dilution with 0.5M sucrose solution. Embryonic survival was estimated from the normal development of embryos for 12, 24 and 48hrs culture. Then the embryos were stained and their cell nuclei was counted. The survival rates of morphological embryos were significantly higher in group I (10% glycerol) or group IV (10% glycerol+10% egg yolk+0.5M trehalose) than those in other groups, although the nuclei number was quite higher in embryos treated with 10% glycerol and 0.25M trehalose at expended blastocyst. However, hatched blastocysts showed higher viability and nuclei number treated with either egg york or trehalose, but the survival rates after 48hrs of culture were quite low. These results indicate that egg yolk and trehalos as a supplement to freezing solutions can be useful to the cryopreservation ofn porcine embryos.

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Effects of the Improvement of Vitrification Solution and FDA-test on the Embryo Survival and Conception Rate by Ultrarapid Freezing II. Effects of the Addition Level of Non-permeable Cryoprotectants (Ficoll, sucrose) in Vitrification Solution and Equilibration Time on the Survival of Vitrofied Mouse Embryos (초급속동결에 있어서 Vitrification Solution 개발과 FDA 생사판정이 수정란의 배양과 이식후 착상에 미치는 영향 II. Vitrification Solution내의 비수과성 물질(Ficoll, sucrose)과 평형시각이 초급속동결 융해후 Mouse Morulae의 생존율에 미치는 영향)

  • 김중계;강민수;장덕지;고경래;양병철
    • Korean Journal of Animal Reproduction
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    • v.16 no.4
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    • pp.317-323
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    • 1993
  • This study was carried out to study effects of the addition level of acetamide and non-permeable cryoprotectants(Ficoll, sucrose) in VS(20% glycerol+10% ethyleneglycol) and equilibration time on the survival of vitrified mouse morulae. The results are summarized as follows: 1. When 10, 15 and 20% of acetamide were added to the new vitrification solution(20G 10E), FDA-scores of embryos were 4.4(control), 4.4(10%), 3.6(15, 20%), respectively. The addition of acetamide did not affect the survival of forzen-thawed morulae(P<0.05). 2. The survival rate betwen 5 min(3.5) and 10 min(4.6), 10 min(4.6) and 20 min(3.2) of equilibration in 10% sucrose, and 20 min(3.2) and 5 min(4.0), or 10 min(4.3) in 20% sucrose were significantly different(P<0.05). The highest survival(4.6) rate was obtained in mouse morulae equilibrated in VS(20G 10E) containing 10% sucrose for 10 minutes. 3. FDA-score of morulae frozen in the new vitrification solution containing 0, 10, 20 and 30% Ficoll was 4.5, 4.2, 4.4 and 4.6, respectively and had no significant effect among concentrations of Ficoll(P>0.05). The development rate after culture(24h) was 89%(20% Ficoll) and 93%(30% Ficoll), respectively.

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A Comparative Study on Fresh and Frozen Embryo Transfer after Superovulation in Black Bengal Goats(Capra-hircus)

  • Mishra, O.P.;Pandey, J.N.;Gawande, P.G.
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.91-100
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    • 2002
  • The experiment was divided into two phases. In phase-I fresh embryos were transferred and in Phase-II frozen embryos were transferred. Embryos were collected by using Dulbecco's phosphate buffered saline. In phase-I total of 65 ova were collected out of 107 ovulation in 18 goats. Recovery of ova was 60.74%, of which 51 (78.46%) was fertilized. Sixteen embryos were transferred to 10 recipient goats and kidding was observed in 6 goats, that produced 10 kids. Thus, 62.50% embryo survival and 60% kidding were achieved in phase-I. In phase-II of the experiment, 17 regular cyclic Black Bengal goats were used. The main purpose was to study the viability of caprine embryos after cryopreservation. In this phase the embryos were collected and frozen using Bio-cool freezers. A two step addition of cryoprotectants (5% glycerol and 10% glycerol) and three-step dilution of cryoprotectants with 1mole (M) sucrose was used. Embryos were preserved for 10 to 45 days. Out of 27 embryos preserved, 18 were recovered after freezing and thawing (37$^{\circ}C$ water bath) with 33.33% embryonic loss. Seventeen frozen and thawed embryos were transferred in 9 recipient goats, out of which kidding was observed in 6 goats and 7 kids were produced, giving a 66.66% kidding and embryo survival of 41.17%. The technique utilized for fresh and frozen embryo transfer can be successfully utilized to produce goats of superior genetic merits. The protocol used for addition of cryoprotectant, freezing, thawing and dilution was found suitable for caprine embryo freezing.

A Modified Cryopreservation Method of Psychrophilic Chlorophyta Pyramimonas sp. from Antarctica

  • Hong, Sung-Soo;Lee, Soo-Young;Kim, Young-Nam;Kang, Sung-Ho;Kim, Hak-Jun
    • Ocean and Polar Research
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    • v.33 no.3
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    • pp.303-308
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    • 2011
  • Polar psychrophiles which thrive under extreme conditions such as cold temperature, high salinity, and high dose ultraviolet light, emerge as novel targets for biotechnology. To prevent genetic drift and the possibility of contamination by subculturing, cryopreservation was employed for two psychrophilic microalgae, Porosira sp. (KOPRI AnM0008) and Pyramimonas sp. (KOPRI AnM0046), which have anti-freeze activities. Five cryoprotectants (dimethyl sulphoxide, ethylene glycol, glycerol, methanol and propylene glycol) showed toxicity at 20-30% (v/v). The optimal cryoprotectant concentration and equilibration time were less than 20% and 10 min, respectively. Cryopreservation was carried out in the presence of cryoprotectants either by direct freezing in liquid nitrogen ($LN_2$) or controlled freezing using a controlled rate freezer followed by storage in the $LN_2$ tank. As a result, Pyramimonas sp. (KOPRI AnM0046), a psychrophilic chlorophyta was revived. Cryopreserved Porosira sp. was not revived from either freezing protocols probably due to the silicic cell wall and its relatively large cell size. In the case of Pyramimonas sp. (KOPRI AnM0046), the controlled freezing method showed higher revival yield than the direct freezing method.

The Effect of Equilibration Temperature and Exposure Time on the Ultrarapid Freezing of 1-cell Mouse Zygote (생쥐 1-세포기배의 초급속 동결에 있어서 평형 온도와 노출시간의 영향)

  • Chung, Duk-Soo;Kim, Hyung-Kuk;Park, In-Kook
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.3
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    • pp.261-268
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    • 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.

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