• Title/Summary/Keyword: 동결보존

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The Effect of Modified Cryopreservation Method on Viability of Frozen-thawed Blastodermal Cells on the Korean Native Chicken(Ogolgye Breed) (한국재래닭(오골계)종 배반엽세포에 있어서 동결 방법의 개선이 융해 후 생존율에 미치는 영향)

  • Kim, Hyun;Kim, Dong-Hun;Park, Soo-Bong;Choi, Seong-Bok;Ko, Yeoung-Gyu;Kim, Jae-Hwan;Do, Yoon-Jung;Park, Hae-Geum;Kim, Sung-Woo
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.65-70
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    • 2012
  • For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at $-7^{\circ}C$ to $-35^{\circ}C$ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen-thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/0.5 ml, and 43.6 % frozen at 20 embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).

레토르트 파우치 해조면의 품질안정화를 위한 최적살균조건

  • 이용우;염동민;여생규;이동호;김동수
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.136-137
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    • 2001
  • 젤리형 해조면류의 유통 및 저장 중에 면가락이 건조되거나 동결하였을 경우 수분이 조직으로부터 분리되어 면의 굵기가 가늘어지면서 딱딱해져 식용하기에 부적당하게 된다. 따라서 이러한 문제를 해결하기 위해서는 먼저 포장을 할 때 적당량의 보존수를 첨가하여 저온 유통을 하여야 한다. 보존수의 첨가 후 저온 유통이 잘 유지될 경우에는 저장 6개월까지는 품질안정성에 문제는 없다. (중략)

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Freezing-and-Thawing Resistance and Strain Characteristics of Recycled Concrete (재생콘크리트의 동결융해저항성과 변형특성)

  • 김광우;이봉학;도영수
    • Magazine of the Korea Concrete Institute
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    • v.4 no.4
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    • pp.115-122
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    • 1992
  • 폐쇄 폐콘크리트를 재활용한 재생콘크리트의 강도특성을 천연골재를 사용한 일반콘크리트와 비교하였다. 동결융해 처리수의 압축강도와 3점휨 재하시험하의 변형율을 측정하였다. 재생콘크리트는 동결융해 처리 후 압축강도 보존율이 더 높았다. 재생콘크리트는 또한 높은 변형율과 처짐에 민감함을 보였으나 파괴와 관련된 다른 성질들은 일반콘크리트와 유사하거나 더 좋은 것으로 나타났다. 그러므로 폐콘크리트를 구조용 콘크리트 제조에 재 사용이 가능할 것으로 보여진다. 그러나 실제 사용을 위하여는 콘크리트에 있어서 중요한 성질인 압축강도가 더 증진되어야 하며 최대 변형율도 보다 자세히 점검되어야 한다.

Study on the Freezing of Bovine Embryos II. Bovine embryos survival after freezing and thawing (우수정란의 동결보존에 관한 연구 II. 동결보존후 융해난자의 생존성)

  • 김정익;양부근;남상헌;이상영;임석기;고광두
    • Korean Journal of Animal Reproduction
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    • v.9 no.1
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    • pp.36-39
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    • 1985
  • This experiment was carried out to investigate the morphology of bovine embryos thawed after deep freezing at -196$^{\circ}C$ and the development of frozen-thawed embryos after in vitro culture in Ham's F-10 medium with 10% NBCS. The results obtained were summarized as follows: 1. The propotion of embryos which a, pp.ared mophologically normal was averaged 77.5% (79/102). 2. The morphologically normal rate of frozen-thawed blastocyst (78.6%) was higher than that of morula (76.7%), but there was no significant difference. 3. Normal development was observed in 20 of 68 embryos cultured for 24-72hr in medium and overall survival rate was 29.4%. 4. Survival rate fo blastocyst (33.3%) was higher than that of morula (25.7%).

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Comparison of AndroMed and Triladyl Extender for Freezing of Jeju Native Black Bull Semen (제주흑우 정액 동결을 위한 AndreoMed와 Triladyl 희석제의 동결성 비교)

  • Cho, Sang-Rae;Cho, In-Chel;Han, Sang-Hyun;Kang, Tae-Young;Cho, Won-Mo;Chae, Hyun-Seok;Kim, Nam-Young;Park, Yong-Sang;Kang, Yong-Jun;Kim, Young-Hoon;Ko, Yeoung-Gyu;Kim, Hyun-Jong;Ko, Moon-Suck
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.155-159
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    • 2013
  • This study was carried out to investigate synthetic extender for semen cryopreservation of Jeju Native Black Bull. The semen was collected using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by Tris-Egg yolk extender and contrifuged in 1,500 rpm for 15 minutes. The supernatant was removed. The pellect was diluted to final sperm concentration of $2{\times}10^8/ml$ by doubling in every 30 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 4 hours at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm for 10 minutes. The height and duration affect the freezing speed by temperature. The frozen straw was plunged to $LN_2$. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. Frozen-thawed sperm were evaluated sperm viability, membrane integrity and acrosome integrity. Post-thawed sperm viability has been significantly higher (p<0.05) in fresh sperm ($93.27{\pm}1.62%$) than frozen-thawed sperm ($73.34{\pm}3.27%$). However, there were no significant differences between fresh and frozen-thawed dead cell rate ($7.35{\pm}2.63$ vs, $13.71{\pm}2.85$). In sperm motility, between Triladyl and AndroMed Extender, there was no significant different ($72.86{\pm}2.83$ vs, $81.47{\pm}2.48$), similarly, the dead cell rates was similar ($18.41{\pm}3.42%$ and $17.26{\pm}4.25$). The results of our study suggest that AndroMed to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Jeju Native Black bull semen.

Studies on In Vitro Fertilization of Follicular Oocytes of Canine using Fresh and Frozen Epididymal Sperm (개 신선 및 동결 정소상체 정액을 이용한 체외수정에 관한 연구)

  • Kim S. K.;Lee B. K.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.301-306
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    • 2004
  • 본 연구는 개의 신선 및 동결 정소상체 정액과 정소상체 정액을 saline가 tris-buffer액으로 희석하고 원심분리에 의해 정장성분을 제거한 정액의 성상과 및 동결보존 시의 생존성 및 난포란과 정소상체 정자의 체외수정 후 체외수정율과 분할율에 대해 조사하였다. 개 정소상체 정액을 saline으로 희석한 정액을 20분간 배양했을 때 정자농도는 3.50$\pm$0.80${\times}$l0$^{6}$ cells/$m\ell$, 정자의 활력은 72.45$\pm$4.55%, 기형정자 수는 7.40$\pm$1.20%로 나타났으며, 대조군인 사출정액의 정자농도는 5.45$\pm$0.50${\times}$$10^{6}$cells/$m\ell$, 정자의 활력은 92.55$\pm$4.65%, 기형정자 수는 3.25$\pm$0.45%와 비교할 때 정자농도와 활력은 낮았으며, 기형정자 수는 많았다. 개 정소상체 정액과 tris-buffer로 희석한 정액을 20분간 배양했을 때 정자농도는 3.80$\pm$0.36${\times}$$10^{6}$ cells/$m\ell$, 정자활력은 78.45$\pm$3.50%, 기형정자 수는 5.54$\pm$0.85%였으며, 희석하지 않은 정소상체 정액의 성상에 비해 약간 높게 나타났다. Tris-buffer로 희석한 개 정소상체 정액을 동결 융해했을 때 생존율은 57.50$\pm$4.20%, 활력은 52.70$\pm$5.50%였으며, 희석하지 않은 대조군의 생존을 74.50$\pm$6.25%와 활력 78.50$\pm$5.20%에 비해 현저히 높게 나타났다. 개 난포란과 신선 몇 동결 정소상체 정자와 체외수정시켰을 때 체외수정율은 각각 63.10$\pm$6.45%, 49.50$\pm$4.28% 및 42.84$\pm$5.90%, 22.30$\pm$5.60%로서 신선 정자에 비해 동결 정소상체 정자로 수정시킨 군의 분할율이 유의하게 낮았다.

Developmental Rate of Vitrified Porcine Oocytes and Its Application to NT Embryos Constructed by Microinjection of Fibroblast Cells into Vitrified Oocytes (돼지 동결 난포란과 이를 이용한 핵이식 배의 체외발생에 관한 연구)

  • Lee, M.-H;Lee, B.-K;Kim, S.-K
    • Journal of Embryo Transfer
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    • v.20 no.3
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    • pp.207-215
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    • 2005
  • 본 연구는 돼지 난포란의 동결보존 후 생존성과 난자의 활성화 처리에 따른 체외발생율과 이를 이용한 핵 이식배의 체외발생율을 조사하였다. 활성화 처리된 배는 $5\%$ FBS가 첨가된 NCSU 23 배양액으로 $38.5^{\circ}C$, $5\%\;CO_2$$95\%$ air의 조건으로 배양하였다. 1. 난포란을 EDS와 $5\%$ PVP로 동결 후 $10\%$ FBS가 첨가된 NCSU 23 배양액으로 $0{\~}10$시간 배양했을 때 체외발생율은 $36.0\%$로서 대조군인 비동결 난포란의 체외발생율 $46.0\%$에 비해 낮았다. 2. Ethanol과 cyclojexamide로 처리 후 42 및 46시간 배양한 배의 분할율은 각각 $33.3\%$, $36.0\%$$27.1\%$, $30.0\%$로서 대조군의 $8.8\%$, $11.4\%$에 비해 높게 나타났다. 3. 동결 및 비동결 난포란을 이용한 핵이식 배의 융합율과 발생율 간에는 유의한 차이가 없었다. 4. Ethanol과 cyclojexamide로 활성화 처리한 난자를 이용하여 재구축한 핵 이식배의 발생율은 $2.8\%$, $5.3\%$$1.5\%$, $2.9\%$로서 대조군의 $0.0\%$, $0.0\%$에 비해 높은 발생율을 나타냈다.

Comparison on Vitrification of Mouse Oocytes and Embryos Using Closed Pulled Straws (CPS), Conventional Straws and Open Pulled Straws (OPS) (CPS, 기존 Straws, OPS 방법을 이용한 마우스 성숙난자 및 수정란의 유리화 동결 비교)

  • Seok H.B.
    • Journal of Embryo Transfer
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    • v.21 no.1
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    • pp.53-58
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    • 2006
  • This study was conducted to comparing on vitrification of mouse oocytes and embryos using CPS, conventional straws and CPS by evaluating in morphological survival for oocytes, and embryonic cleavages and blastocyst formation for embryos. The morphological survival in vitro after thawing of vitrified oocytes using CPS (75%) and conventional straws (72%) were significantly higher (p<0.05) than that using OPS (68%). The blastocyst formation rates of vitrified embryos using CPS (48.6%) and unfrozen control embryos (56.0%) were significantly higher (p<0.05) than those of conventional straws (43.4%) and OPS (37.7%). The rates of morula formation were also higher to control, CPS, conventional straws and OPS in orderly. These results show that CPS has the advantages of achieving a high survival and safety preservation.

Improved Ultrastructural Preservation of Retinal Cells in Drosophila melanogaster (초고압동결장치를 이용한 초파리 레티나 세포의 향상된 미세구조)

  • Mun, Ji-Young;Park, Se-Jin;Han, Sung-Sik
    • Applied Microscopy
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    • v.37 no.3
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    • pp.175-183
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    • 2007
  • The Drosophila retinal cell is widely used to study cell development and cell signaling processes. In the past decades, conventional chemical fixation had been used to study the structure of retinal cells in Droscphila. Rapid freezing methods are superior to chemical fixation methods due to their fixation speed. Some Drosophila tissues, such as the eyes, should not be freezed due to their surrounding cuticle layer. Therefore, in the case of the Drosophila retina, the benefits of high pressure freezing and freeze substitution (HPF-FS) had not been verified. In this study, a retinal cell from Drosophila melanogaster had been studied by using the HPF-FS method. Compared to chemical fixation, the preservation of the cytoplasm in the HPF-FS sample was improved on the whole. The HPF-FS cell membranes were smoother than that of chemical fixation. In addition, HPF-FS preserved the mitochondria structures very well. These results of the present study suggest that HPF-FS is superior to other fixation methods for the preservation of the retinal cell structure.

Conservation Treatment for Wooden Frame of Armor Excavated From Im-dang, Gyeong-san (경산 임당출토 목제 갑옷틀 보존)

  • Yi Yong-Hee;Kim Soo-Chul
    • 한국문화재보존과학회:학술대회논문집
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    • 2005.11a
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    • pp.246-251
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    • 2005
  • 경산 임당유적에서 출토된 목제 갑옷틀은 수침된 상태로 출토된 목제유물로 재질이 매우 취약하여, 대기 중에 노출되면 급속도로 건조가 진행되어 곧바로 수축 변형이 일어나 유물로써 가치를 잃어버리게 된다. 특히 목제 갑옷틀은 오랜 매장기간동안 목질부의 분해가 많이 진행되어 재질이 취약할 뿐 아니라 원통목을 가공한 것이어서 건조 과정에서 수축변형이 발생우려가 높다. 따라서 목제 갑옷틀은 건조변형이 적고 치수안정화가 우수한 처리법을 적용하여야 한다. 따라서 목제 갑옷틀은 t-butanol 용액을 용제로 PEG#4000 40%로 함침 전처리 후 진공 동결건조법으로 보존처리 하였다.

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