• Title/Summary/Keyword: 동결 시간

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The Use of Styrofoam Box for Chikso (Korean Brindled Cattle) Semen Cryopreservation with Liquid Nitrogen (칡소 동결 정액 생산을 위한 스티로폼상자와 액체질소 이용 방법)

  • Kim, Sung Woo;Ko, Yeoung-Gyu;Lee, Jae-Yeong;Kim, Chan-Lan;Hwang, In-Sul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.4
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    • pp.490-496
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    • 2020
  • A styrofoam box is used as a simple and easy freezing method to preserve animal semen as a livestock genetic source. This study optimized the methods of freezing chikso brindled cattle semen. To test the freezing box, the motility of spermatozoa was compared between two box sizes (length×width×heigh) with the dimensions of 23.5×30.5×22.5 cm and 25.5×46.5×26.5 cm. The motility of thawed sperm from brindled Korean bulls was used to confirm the efficiency of the freezing boxes. The box having a larger inner space with larger horizontal and height measurements supported better motility after thawing (60.4±5.3% vs 67.2±3.1%) with 10 min of exposure time in liquid nitrogen vapor. The optimized freezing space is estimated to be an essential element for successful freezing results and the larger box could be used for production of more than 60 frozen semen straws. These properties are also helpful to optimize the cryopreservation techniques that would control the quality and quantity of semen straws according to different animal species.

Effects of Cooling Rate and Equilibration Time on the Survival and Development of Frozen-thawed Bovine Immature Oocytes (동결속도와 평형시간이 소 미성숙 난자의 동결 융해후 생존율에 미치는 영향)

  • 양병철;양보석;성환후;임기순;최선호;장원경;진동일;임경순
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.19-28
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    • 2001
  • The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

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Effects of Cryopreservation on the Seed Germination and Growth Properties of Seedlings of Maackia amurensis (초저온 저장이 다릅나무 종자의 발아와 유묘의 생장특성에 미치는 영향)

  • 한심희;김찬수;장석성;이현주
    • Korean Journal of Plant Resources
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    • v.17 no.2
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    • pp.75-81
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    • 2004
  • This study was conducted to investigate effects of cryopreservation by vitrification on the seed germination rate and growth and physiological properties of seedlings of Maackia amurensis. Cryopreservation significantly decreased the germination rate of seeds of M. amurensis, but the reduction of germination rate was mitigated by the treatment of cryoprotectant (plant vitrification solution, PVS2) before plugging into liquid nitrogen and fast thawing rate after cryopreservation. Long-term PVS2 exposure decreased seed germination rate, whereas cryopreservation time didn't have influence on seed germination rate. In addition, growth and physiological properties of seedlings were not affected by PVS2 exposing time and cryopreservation time. Therefore cryopreservation could be widely used as a technique of long-term ex situ conservation without any damage and deterioration of cells or tissues of the forest seeds. However, in order to increase the effect of cryopreservation, we have to develope the lower toxic cryoprotectant and suitable techniques to the structural or chemical properties of a variety of seeds.

Effect of Freezing on Hydration Rate and Sprouting Characteristics of Soybeans (대두의 수화속도와 콩나물의 발아특성에 대한 동결처리 효과)

  • Rhim, Jong-Whan;Oh, Bong-Yun;Kim, Dong-Han;Ham, Kyung-Sik
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.660-665
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    • 2003
  • Effect of freezing on the hydration rate and growth characteristics of bean sprout of two domestic cultivars of soybean, Eunha beans and Taegwang beans, was investigated. Freezing of the soybeans at $-80^{\circ}C$ affected characteristics of the hydration and growth of soybean sprout of the beans. Hydration rate of the frozen beans depended on the hydration temperature. It decreased up to 50% at lower than $60^{\circ}C$ of hydration temperature, but it did not decrease at higher than $60^{\circ}C$. Activation energies of frozen Eunha and Taegwang beans were 39.79 and 39.25 kJ/mol, respectively. Kinetic compensation effect between activation energy values and the pre-exponential factor for the hydration of soybeans with or without freezing was observed. Germination rate and thickness of the bean sprout increased by freezing, however, yield and weight were not affected by freezing.

Cryopreservation of Mouse IVF/IVC Blastocysts by Vitrification (체외수정된 생쥐 배반포기배의 초자화 동결)

  • Kim, Seun-Eui;Uhm, Sang-Jun;Kim, Eun-Young;Yoon, San-Hyun;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.1
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    • pp.41-49
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    • 1996
  • 본 연구는 체외수정에 의해 생산된 생쥐 배반포기배를 vitrification 방법으로 동결보존하였을때 높은 생존율을 얻기 위한 적정조건을 검토하고자 실시하였다. 배반포기배를 생산하기 위하여, B6CBA F1 (C57BL/6, (표현불가)${\times}CBA/N$, (표현불가)) 계통의 생쥐 미수정란에 $1{\times}10^6$ spermatozoa/ml 농도의 정자로서 수정을 유도하였으며, 이후 $37^{\circ}C$, 5% $CO_2$배양기내에서 96시간동안 체외배양하였다. 배양 4일째의 배반포기배는 발달상태에 따라 early, middle 그리고 hatching blastocysts로 구분하였다. 본 실험에 사용된 동결보존액은 30% Ficoll과 0.5mol의 sucrose가 첨가된 mDPBS 용액에 40%의 ethylene glycol를 첨가한 EFS 40 (Zhu et al., 1993) 이었고, 수정란은 $25^{\circ}C$의 상온에서 먼저 20% ethylene glycol에 노출된 후 EFS 40 용액으로 옮겨 액체질소에 침지하는 2단계 동결법에 의해 동결보존되었으며, 급속융해하여 다음과 같은 결과를 얻었다. 1. 체외수정율과 배양 4일째 배반포기까지의 배발달율은 각각 89.4%와 86.1%였다. 2. 20% ethylene glycol에서 5분간 평형된 후 EFS 40 용액에 냉동보존된 후 융해된 난자의 생존율은 20% ethylene glycol에 0, 1, 3분간 평형된 난자의 생존율에 비해 유의하게 높았다. 3. 배반포기배를 20% ethylene glycol에서 5분간, EFS 40 용액에 1분간 차례로 노출한 다음 체와배양하였던 바, 배양 24시간째 생존율은 $82.9%{\sim}88.4%$ 였다. 본 연구 결과, 체외수정, 배양된 생쥐 배반포기배는 20% ethylene glycol과 EFS40에 대한 노출만으로는 난자의 생존성에 나쁜 영향을 미치지 않는 것으로 미루어 보아 배반포기배의 초자화 동결이 가능함을 시사하였다. 따라서 동결 융해 후 높은 생존율은 상온에서 난자를 2단계 즉, 20% ethylene glycol에 5분간 평형시킨 후 EFS 40 용액에 노출하여 1분내에 LN2에 직접 침지하는 간편한 동결방법으로 얻을 수 있었다.

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Numerical Investigation into Behavior of Retaining Wall Subject to Cycles of Freezing and Thawing (동결-융해 반복작용에 노출되는 옹벽의 거동에 관한 수치해석 연구)

  • Yoo, Chung-Sik
    • Journal of the Korean Geotechnical Society
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    • v.29 no.1
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    • pp.81-92
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    • 2013
  • This paper presents the results of a numerical investigation into the behavior of retaining wall subject to cycles of freezing and thawing due to seasonal temperature change. The thermo-hydro-mechanical coupled finite element modeling strategy was first established to simulate the wall behavior. A series of finite element analyses were then performed on a range of conditions representing seasonal temperature change characteristics. The results indicated that the average freezing temperature and the number of cycles of freezing and thawing were the primary influencing factors for the wall behavior. Also revealed was that the duration of freezing period does not significantly affect the wall displacement and the lateral earth pressure, and that the earth pressure on the wall does not significantly change due to the freezing and thawing action suggesting that the increase in the wall displacement during the freezing and thawing action may be attributed to degradation of backfill due to the freezing and thawing action.

Cryopreservation of Recipient Oocytes Collected from Korean Native Cattle: Incidence of the Zona Hardening of Recipient Oocytes Collected from Korean Native Cattle at the Different Stages of Cryopreservation (복제 한우 생산을 위한 수핵난자의 동결에 관한 연구 : 상이한 동결과정 중 한우 수핵난자에서 일어나는 투명대 경화)

  • 이병천;박종임;임정묵;이은송;노상호;황우석
    • Journal of Embryo Transfer
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    • v.14 no.2
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    • pp.99-106
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    • 1999
  • 핵 이식에 공여되는 수핵난자의 효과적인 동결보존을 위하여 한우 성숙난자를 1.0 M dimethylsulfoxide(DMSO) 또는 1.0 M glycerol이 함유된 동결보호제를 이용하여 처리하거나, 동결보호제 처리 후 완만동결법을 이용한 동결융핼르 시행하여 상기 실험처리로 야기되는 투명대 경화현상을 관찰하였다. 도축장 유래의 난소에서 미성숙난자를 채취한 후 10% 소 태아혈청을 함유한 TCM-199을 이용하여 22∼24 시간 동안 체외성숙배양을 이해하였다. 배양후 작출된 성숙난자를 각각의 동결보호제로 처리, 혹은 처리 후 동격융해한 후 protease를 이용하여 투명 대의 경화현상 발생의 빈도를 조사하였다. 또한 동격란을 동결정액을 이용한 체외수정에 공여한 후 정자 침입농도능을 조사하였다. 동결보호제로 처리한 난자에 있어서 보호제의 종류와 관계없이 투명대 경화현상이 유의적 (P<0.05) 으로 증가하였으나 이후의 동결융해 처리에 의한 추가적인 경화현상의 발생은 증가하지는 않았다. 또한 투명대 경화현상의 발생양상을 동결보호제 처리 후 10분 간격으로 측정한 결과 DMSO의 경우 처리후 10분, glycerol의 경우 처리 후 20분 후부터 유의적으로 차를 발견할 수 없었으며, 수정율 및 난자 1개당 침입한 정자의 수는 동결란에서 유의적으로 증가하지 않았다. 본 연구의 결과 동결난자의 투명대 경화현상은 동결보호제 처리과정에서 이미 일어나지만, 이러한 투명대 경화현상이 난자의 동결보존 후 수정능에는 현자한 영향을 미치지 않는다는 사실이 규명되었다.

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한우의 개체별 체외수정란 생산

  • 임여정;정연길;석상현;박성백;송해범
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.139-139
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    • 2003
  • 최근, 한우의 수정란 체외이식두수가 급격히 늘어나고 있으나, 모축의 등록번호를 대부분 확인 할 수 없으며, 확인이 가능할지라도 개체별 수정란의 생산이 거의 이루어지지 않고 있다. 본 실험에서는 모축의 등록번호가 확인된 개체의 난소로부터 채취한 난포란만을 체외성숙, 수정, 배양하여 배 발생율을 조사하고, 동결성적을 조사하였다. 도축된 한우의 난소를 개체별로 채취하여 2$0^{\circ}C$생리식염수에 넣어 3시간 이내에 실험실로 운반하고, 난포란을 채취하였다 개체별로 채취한 난포란은 9~47개/난소였고, 평균 14.3개/난소였다. 개체별 난포란은 IVMD 101 (일본, 펩티트연구소)에서 22물24시간 동안 체외 성숙시킨 후, IVF 100(일본, 펩티트연구소)으로 2회 세정한 후, 각각의 배양액 50${\mu}\ell$ 소적에 5개씩 5~6시간 수정시켰다. 수정란을 저 산소 배지 IVD 101 (일본, 펩티트연구소)로 7~9일간 배양하여 배 발생율을 조사하고 동결하였다. 평균 수정율은 81%, 배반포의 발생율은 35% 였으며, 동결 가능한 수정란은 평균 4개/두 정도 얻을 수 있었다. 이상의 결과로 한우의 개체가 확인된 난소로부터 수정란생산이 가능하고, 육질등급이 우수한 한우의 수정란생산이 가능해졌으며, 앞으로 고 능력 젖소의 난소에서도 고능력 수정란의 생산이 가능할 것으로 사료된다.

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Effects of Salt Concentration on the Rehydration Characteristics of Freeze Dried Mook (재수화용액의 염농도에 따른 동결건초 도토리 묵의 재수화 특성)

  • 윤광섭;황정섭;정헌식;양경미
    • Food Science and Preservation
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    • v.8 no.3
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    • pp.313-319
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    • 2001
  • In order to produce the high quality rehydrated acorn Mook(Korean tradition gel flood) that enhance acceptability, the optimum condition was investigated for the rehydration process of dried Monk as salt concentration(0, 1, 2%), temperature(20, 70, 80, 90$^{\circ}C$) and time. The estimation of moisture gain, rehydration efficiency was analyzed statistically. The surface color md seniory evaluation were undertaken to evaluate the rehydrated Mook quality The optimum rehydration time was decided to 15 minutes and it takes 3 minutes for the cooling tilde. The moisture gain increased as the rehydration temperature increase. And the moisture gain and moisture gain rate were higher at 1% salt solution than other concentration. As the rehydration efficiency, surface color and sensory properties of rehydrated Mook, 1% salt treatment was superior.

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Changes of Volatile Odor Components in Kimchi by Freeze-drying (동결건조에 의한 김치의 휘발성냄새성분의 변화)

  • Ko, Young-Tae;Kang, Jung-Hwa
    • Korean Journal of Food Science and Technology
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    • v.34 no.4
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    • pp.559-564
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    • 2002
  • Volatile odor components of ripened and ripened/freeze-dried kimchi were analyzed by gas chromatograph. As ripening temperature of kimchi increased, pH of kimchi decreased, viable cell count of lactic acid bacteria of kimchi increased up to ripening temperature of $15^{\circ}C$, and sensory properties of kimchi gradually decreased. Allyl mercaptan, methyl allyl sulfide, dimethyl disulfide, diallyl sulfide, diallyl disulfide, and ethanol were detected in ripened kimchi and ripened/freeze-dried kimchi. The amounts of allyl mercaptan, methyl allyl sulfide, diallyl sulfide, and ethanol increased as the ripening temperature increased, while those of dimethyl disulfide and diallyl disulfide decreased. Freeze-drying for 24 hr removed most of the above-mentioned volatile odor components, which were further removed by freeze-drying for 48 hr.