• Title/Summary/Keyword: 동결방법

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MICROVENOUS ALLOGRAFTS IN RABBIT FEMORAL ARTERIES : EXPERIMENTAL STUDY OF FREEZE-DRIED ALLOVEINS (동종 정맥 이식을 이용한 가토 대퇴동맥 결손부 수복(동결건조 동종정맥이식의 실험적 연구))

  • Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.13 no.4
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    • pp.355-369
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    • 1991
  • 미세수술을 이용한 재건외과 분야에서 빈번히 혈관이식이 요구된다. 이러한 경우 자가정맥이 가장 널리 사용되고 있으며 그의 신뢰성도 인정되어 있다. 그러나 정맥 채취에 따르는 부가적인 수술이 요구되며 혈관 공여부에 또다른 결손을 초래한다. 동결건조동종정맥은 이러한 점을 보완하고 자가정맥을 대체할 수 있는 잠재성이 있다. 이에 동결건조동종정맥의 효율성을 알아보고자 2.5cm 길이의 가토대퇴정맥을 $-65^{\circ}C$, 200 mtorr의 음압으로 동결건조시킨 다음 대퇴동맥 결손부에 동종이식하고(n=24), 신선한 가토대퇴정맥 동종이식군(n=24)과 자가정맥이식군(n=24)을 1주 간격으로 4주간 비교 관찰하였다. 2주 개존율은 동결건조동종정맥 이식군, 100%;동종정맥이식군, 50%; 자가정맥이식군, 100%이었으며 4주 개존율은 동결건조동종정맥이식군, 83.3%;동종정맥이식군, 50%;자가정맥이식군, 100%로서 동결건조처치만으로 동종정맥이식의 생존율을 증가시켰다. 미세임파구세포독성검사에서는 동결건조정맥의 항원성이 상당히 낮아져있음을 알 수 있었다. 그러나 동결건조정맥의 내막세포화가 주사전자현미경 및 광학현미경 소견상 자가정맥보다 지연됨이 관찰되었다. 이러한 결과를 종합해 볼 때 동결건조동종정맥은 아무런 처치를 하지않은 신선 동종정맥보다 현저한 장점이 있었지만, 자가정맥이식을 대신할 수 있는 보다 더 좋은 대체방법이라는 견지에서는 임상 적용이 어려울 것으로 사료되었다.

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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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Chromosome and Spindle Configuration of Mouse Oocytes after Vitrification at the Mature Stage (마우스 성숙난자의 유리화 동결법에 따른 동결 융해 후의 염색체와 방추사의 분석)

  • ;;;;Gary B. Anderson
    • Korean Journal of Animal Reproduction
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    • v.25 no.3
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    • pp.287-292
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    • 2001
  • Selection of oocyte cryopreseivation method is a prerequisite factor for developing an effective bank system. To develop an effective vitrification method, we examined whether damages in spindle and chromosome morphology induced by vitrification. Intact cumulus-enclosed oocytes were vitrified with DPBS with 5.5 M ethylene glycol and 1.0 M sucrose, and loaded onto eletron microscopic copper grid for storing in liquid nitrogen. Intact vitrified and thawed oocytes were immunostaining for tubulin and karyotying for chromosome. Vitrfied and thawed oocytes had a higher rate of chromosome (32.8% vs. 19.6%) and spindle (32.3% vs. 20.2%) abnormalities compared with fresh oocytes. Mouse oocytes after vitrification at the mature stage showed increased incidence of chromosomal and spindle abnormalities.

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Extraction Characteristics of Rubi Fructus in Relation to Drying Methods and Extraction Solutions (복분자의 포제방법에 따른 추출물 특성)

  • Chung, Hun-Sik;Hwang, Sung-Hee;Youn, Kwang-Sup
    • Food Science and Preservation
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    • v.12 no.5
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    • pp.436-441
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    • 2005
  • This study was observed the effects of drying methods on the extraction characteristics of Rubi fructus (fruits of Rubus coreanus). Extraction yields of soluble solids and total sugar were high in the Rubi fructus dried by freeze drying, followed by infrared drying and sun drying. Extraction yield of phenolic compounds and DPPH radical-scavenging activity of extracts were in the following order; the Rubi fructus dried by freeze drying, the Rubi fructus dried by infrared drying, the Rubi fructus dried by sun drying. L value was the highest in the Rubi fructus dried by freeze drying, and a and b value were lowest in the fruit dried by freeze drying. These results suggest that freeze drying has an beneficial effect to enhance the quality of Rubi fructus. Water and ethanol extractions was more effective in the extraction of soluble solids and the antioxidative components.

Changes of Physicochemical and Sensory Characteristics in the Codonopsis lanceolata Saengsik, Uncooked Food by Different Drying Methods (건조방법 달리하여 제조한 더덕 생식의 이화학적 및 관능적 특성 변화)

  • Jin, Tie-Yan;Quan, Wu-Rong;Wang, Myeong-Hyeon
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.721-725
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    • 2008
  • Changes in the physicochemical and sensory characteristics of the uncooked food, Codonopsis lanceolata saengsik, were investigated to determine an efficient drying method, one of the most important manufacturing processes in the preparation of C. lanceolata saengsik. No changes in the proximate compositions of all samples were observed during hot-air drying at 50 and $60^{\circ}C$ and freeze-drying. The L value in the freeze-dried sample was higher than that in the hot-air dried samples, whereas the b value in the freeze-dried sample was reduced. Dietary fiber content in the hotair dried samples were higher than that in the freeze-dried sample, whereas the total phenolic compounds and crude saponin contents were lower than those in the freeze-dried sample. The highest overall acceptability values in the sensory test for color, flavor, taste, and overall acceptability were 5.63, 5.45, 5.75, and 5.85, respectively. In conclusion, the freezedrying method was the most favorable of the tested method for the manufacture of C. lanceolata saengsik.

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.

Sperm Cryopreservation of Pacific Cod Gadus macrocephalus (대구(Gadus macrocephalus)의 정자 동결보존)

  • Do, Yong Hyun;Cho, Jae Kwon;Lee, Hee Jung;Min, Byung Hwa
    • Korean Journal of Ichthyology
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    • v.31 no.4
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    • pp.195-200
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    • 2019
  • An experiment was performed to obtain cryopreservation techniques of Pacific cod Gadus macrocephalus sperm. Milt were cryopreservation using five cryoprotectant demethyl sulfoxide (DMSO), ethylene glycol (EG), glycerol, methanol and propylene glycol (PG) with marine fish ringer's solution (MFRS) as diluent. Milt were cryopreserved each experimental methods like cryoprotectants (10% and 20%), equilibration time (3, 5 and 10 min) and freezing protocols (liquid nitrogen vapor above 3, 8 and 12 cm). Post-thaw sperm survival rate revealed the highest in 10% PG with optimum methods of equilibration time (3 min) and freezing protocol (liquid nitrogen vapor above 8 cm) about 21.3±1.8%. Hatching rate of fertilization eggs using fresh and cryopreserved sperm were no significantly different.

A Compound Deterioration Assessment of Concrete Subjected In Freezing-Thawing and Chloride Attack (동결융해와 염해의 복합작용을 받는 콘크리트의 내구성능 저하 평가)

  • 고경택;김도겸;김성욱;조명석;송영철
    • Journal of the Korea Concrete Institute
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    • v.13 no.4
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    • pp.397-405
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    • 2001
  • In clod weather regions, a strong seasonal wind brings sea salts to the land. In addition to it, recently, the spreading amount of deicing salts has increased numerously for purpose of removing snow and ice. Thus the salts environment around concrete structures becomes so severe that various damages of concrete due to applied salts will be brought up. Much of countries such as America, Europe etc. is carried out study for effects of deicing salts on concrete. However, there are not test methods for deterioration of concrete subjected to both freezing-thawing and chloride in Korea. In this study, we carried out test for the compound deterioration subjected to both freezing-thawing and chloride attack, to investigate the effects of sodium chloride on the deterioration of concrete. The test was performed to investigate the effects of cement type, strength and air content on the scaling deterioration of concrete. As a result, the scaling deterioration was accelerated in the presence of salts. And the resistance to scaling was strongly influenced by the type of cement, the strength and air content of concrete.

Comparison of Vitrification and Slow Freezing for the Cryopreservation of Chicken Primordial Germ Cell (Ogye) (한국재래닭 (오계) 원시생식세포의 완만동결과 급속동결의 비교)

  • Kim, Sung Woo;Ko, Yeoung-Gyu;Byun, Mijeong;Do, Yoon Jung;Han, Jae Yong;Kim, Dong Hun;Seong, Hwan-Hoo;Kim, Hyun
    • Journal of Animal Science and Technology
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    • v.55 no.5
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    • pp.417-425
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    • 2013
  • We sought to provide a method for freezing and preserving primordial germ cells, or an avian germ cell of a bird, as a material for developmental engineering or species preservation. The aim of this study was to compare the efficacy of slow freezing with a vitrification method for the cryopreservation of chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonad of day 5.5-6 day (stage 28) cultured chick embryos, using the MACS method, were classified into two groups: slow freezing and vitrification. We examined the viability of PGCs after Cryopreservation. Four freezing methods were compared with each other, including the following: Method 1: The PGCs were frozen by a programmed freezer in a plastic straw, including 2.0 M ethylene glycol (EG) as cryoprotective additive (slow freezing) Method 2: The PGCs were vitrified in a plastic straw, including 8.0 M EG, plus 7% polyvinylpyrrolidone (PVP) (rapid freezing). Method 3: The slow freezing was induced with a cryotube including 2.0 M EG Method 4: The PGCs were frozen in a cryotube including 10% dimethyl suloxide (DMSO) (rapid freezing). After freezing and thawing, survival rates of the frozen-thawed PGCs from Method 1 to 4were 76.4%, 70.6%, 80.5% and 78.1% (p<0.05), respectively. The slow freezing ($-80^{\circ}C$ programmed freezer) method may provide better survival rates of frozen-thawed PGCs than the vitrification method for the cryopreservation of PGCs. Therefore, these systems may contribute to the cryopreservation of a rare avian species.

Freezing and Washing of Red Blood Cells Using Haemonetics ACP 215 (Haemonetics ACP 215를 이용한 적혈구의 동결과 세척)

  • Youn, Kyoung Won;Choi, Kyoung Young;Lee, Sun Ah;Min, Hyuk Ki;Kim, Jaehyun
    • The Korean Journal of Blood Transfusion
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    • v.29 no.3
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    • pp.291-300
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    • 2018
  • Background: The use of a functionally closed system for the glycerolization and deglycerolization of red blood cells (RBCs) allows for prolonged post-thaw storage for more than 24 hours. The aim of this study was to assess glycerolization and deglycerolization processing for RBCs using a high glycerol method in the automated, closed system provided by Haemonetics ACP 215. Methods: Thirty-five packed RBCs were glycerolized using the ACP 215 to a final concentration of 40% (wt/vol). The units were either frozen as such (n=30) or excess glycerol was removed (n=5) before freezing. After storage at $-80^{\circ}C$, the units were thawed, deglycerolized and resuspended in SAG-M. The frozen-thawed RBCs were stored at $4^{\circ}C$, and analyzed for their stability and in vitro quality. Results: No prefreeze excess glycerol removal units showed significantly less potassium leakage during post-thaw storage compared to the prefreeze excess glycerol removal units. All measurements of the stability and in vitro quality of thawed RBCs prepared from frozen RBCs without the prefreeze removal of excess glycerol during post-thaw storage at $4^{\circ}C$ for 7 days were acceptable to the American Blood Bank Association's standards and European standards. Conclusion: RBCs frozen without prefreeze removal of excess glycerol and the ACP 215 simplifies cryopreservation procedure and increases the stability of frozen-thawed RBCs. This increases the practical applicability of cryopreserved RBCs in blood transfusion practice.