• 한국가축번식학회지
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• 제16권4호
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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.

• 한국수정란이식학회지
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• 제11권1호
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• pp.41-50
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• 1996

This experiment was carried out to investigate the ovarian responses of the ovulation point, ovarian weight and size, the number of ovarian follicles and collected embryos, and to study the effects of the developmental stages (oocytes, 2-4 cell. 8-16 cell and morulae), additional levels of Ficoll (0, 15, 30%) on the survival rate (FDA-test) of rat embryos frozen in vitrification solution (20% glycerol + 10% ethylene glycol + 10% sucrose). Sunanarized results was as follows; 1. The mean ovulation point per head was 7, and the weight of ovaries was 0.03g. The size of ovary was 5.9 mm(L) and 4.6 mm(W), and the number of ovarian follicles over and below 2 mm was 4.7 and 8.7, respectively. The number of the collected embryos per head was 5.5 (79%). 2. 2. The FDA score of embryos frozen in 20 G 10 E 10 S without Ficoll was 2.8 (oocyte), 2.6 (2-4 cell), 3.9 (8-16 cell) and 3.6 (morula), respectively. However, there were no significant differences among treatments. 3. The FDA score of embryos frozen in 20 G 10 E 10 S with 15% Ficoll was 3.4 (oocyte), 4.0 (2-4 cell), 4.7 (8-16 cell) and 4.8 (morulae), respectively (P>0.05). 4. The FDA score of embryos frozen in 20 G 10 E 10 S with 30 % Ficoll was 3.7 (oocyte), 3.2 (2-4 cell), 4.4 (8-16 cell) and 4.4 (morulae), respectively (P>0.05). 5. As shown in the above results, the higher survival rate was obtained in the treatment of 15% Ficoll than that of 30%. And the survival rate (FDA-test)of the oocytes and 2-4 cell stages of the rat embryos was lower than that of 8~16 cell and morulae stages. It was considered that 8-16 cell and morulae could be available for the successful freezing by vitrification of rat embryos with 15% Ficoll except for oocytes.

• 원예과학기술지
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• 제17권2호
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• pp.138-140
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• 1999

카네이션의 약배양 기술을 확립하고자 저온 전처리, 암처리, Ficoll처리가 약배양에 미치는 효과를 조사하였다. 약배양에 적합한 화분의 발육단계인 1핵기 소포자 전후 화분의 약을 가지는 카네이션 화뢰의 직경은 4~6mm이었다. 카네이션 약배양 후 캘러스 형성과 배양 후 약의 반응에 가장 효과적인 저온전처리 기간은 7일이었다. 명배양하는 것보다 암배양하는 것이 캘러스 형성과, 형성된 캘러스로부터 root 분화에 효과적이었다. Ficoll처리는 캘러스 형성에 큰 효과는 없었지만, 형성된 캘러스로부터 신초와 root가 분화되었다.

• 한국수정란이식학회지
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• 제13권2호
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• pp.191-197
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• 1998

Immature nocytes and in VitrO matured Oocytes collected from the slaughtered Korean cattle were frozen slowly with 10% ethylene glycol+5% polyvinyl pyrolidine+0.05M trehalose (l0EPT), 10% ethylene glycol+5% ficoll+0.05M sucrose (1OEFS), or 10% ethylene glycol+5% ficoll+0.05M trehalose (l0EFT) by cell freezer (experiment 1). And also,They were ultra-rapidly frozen with 30% ethylene glycol+10% polyvinyl pyrolidine+0.5M trehalose (3OEPT) or 30% ethylene glycol+18% ficoll+0.5M sucrose (3OEFS) using electron microscope grid (experiment 2). In experiment 1, the cleavage rate was 23.0% when immature oocytes were frozen slowly using various cryoprotectants descrihed above, and 5.1% of cleaved oocytes developed to over morula stage after in Vitro fertilization (IVF). There were no significant differences among these groups. When matured oocytes were frozen slowly, the total cleavage rate was 19.7%, and over morula stage was 3.2%. lOEPT (4.8%) and EFS (4.4%) were slightly more effective than l0EFT (0.0%) for development in vitro. Only in l0EFT treated group, immature oocytes have higher developmental capacity than matured ones, when they were frozen slowly and IVF after thawing. In experiment 2, oocytes were ultra-rapidly frozen using the electron microscope grid with two kind of cryoprotectants described above. In immature oocyte group, the cleavage rate was 13.9% and 5.8% of cleaved oocytes developed to over morula stage after IVF, and in matured group, 25.7 and 7.6%, respectively. There were no significant differences between two kind of cryoprotectants, but in ultra-rapid freezing using electron microscope grid, the efficiency is slightly higher in matured oocyte group.

• 대한수의학회지
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• 제29권2호
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• pp.33-39
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• 1989

Pure separation of various leukocytes is required for the assessment of their roles in immunological and phisiological function. In this study, pure separation of monocytes from canine peripheral blood was attempted. At first, mononuclear cells (PBMC) were separated by ficoll-hypaque gradient method and then monocytes were recovered from PBMC suspensions in sucrose gradient Sol. (PBMC-Sucrose), autologous plasma (PBMC-Plasma) and autologous serum (PBMC-Serum) incubated at $37^{\circ}C$ for 2 hours. 1. In the separation of PBMC by ficoll-hypaque gradient method in canine blood, higher relative centrifugal force (RCF) was required, as high as more than 1,300xg RCF for 40 minutes, for clear formation of PBMC layer than that in human blood as usually used 400xg RCF for 40 minutes. 2. In monocytes-separation from three PBMC suspensions following PBMC separation, recovery-, purity- and viability-rate of monocytes showed better results in PBMC-Plasma and PBMC-Serum than in PBMC-Sucrose suspension, particulary showing better results from PBMC suspensions performed by centrifugation at 1,500xg RCF for 40 minutes.

• 대한의생명과학회지
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• 제4권1호
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• pp.35-42
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• 1998

치료목적으로 채취한 사람 복수에서 C반응성 단백질 (CRP)을 분리 정제하여 사람 대식세포 탐식활성에 미치는 영향에 대하여 연구하였다. CRP는 p-diazonium phenyl-phosphoryl choline 혹은 C-polysaccharide coupled sepharose 4B와 hydroxylapatite affinity chromatography법으로 분리 정제 시켰다. 대식 세포는 ficoll hypaque 밀도 원심 구배법으로 분리시킨 다음 부착법으로 정제시키고 탐식 시험을 이용하여 확인하였다. CRP가 대식 세포의 시험 관내 미생물 탐식 활성에 미치는 영향은 촉진 혹은 억제시키는 경향을 보였다. 즉, CRP와 대식세포 미생물 탐식능과의 관계는 반응시킨 시간, 반응계에 가해준 CRP량, 미생물ㆍ탐식세포 및 CRP상호간 반응시킨 순서에 따라서 다르게 나타났다. 대식세포의 시험관내 탐식활성에 미치는 CRP 자극의 특성은 한계자극 특성을 보였다.

• 한국가금학회:학술대회논문집
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• 한국가금학회 1999년도 제16차 정기총회및학술발표회
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• pp.11-33
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• 1999

This study was conducted to determine the embryonic stages for the isolation of the highest number of PGCs and to improve PGCs enrichment method. The primordial germ cells(PGCs) from different sources of chick embryos were isolated. The embryonic stage having the highest number of PGCs from each sources was selected ; 1-day-old embryos for germinal crescent (stage 6-8), 2.5-day-old embryos for blood (stage 17-18) and 5.5-day-old embryos for gonad (stage 27-28). The number of PGCs from one embryonic germinal crescent, blood and gonad was about 87$\pm$1.8, 103$\pm$4.0, and 932$\pm$10.9, respectively. The viability of PGCs after Ficoll from each sources was similar, showing approximately 70%. the PGCs enrichment method was improved using Ficoll density gradient centrifugation. After this step the purity of PGCs from germinal crescent, blood, and gonad was 45$\pm$9.10%, 85$\pm$1.18%, and 86$\pm$0.19%, respectively. Also, PGCs were picked up by mouth pipette to improve the purity. This improved method for the separation of PGCs from different sources will serve as a useful too to preserve the foundation stocks of poultry and to produce germline chimeras.

• Archives of Pharmacal Research
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• 제13권4호
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• pp.325-329
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• 1990

Synaptosomal plasma membrane vesicles (SPMV) were isolated from fresh bovine cerebral cortex by several well-known procedures, and the best procedures was selected by enzymatic and morphological standards. The SPMV isolated by the modified procedure of Smith and Loh showed the highest purity. The specific activities of Na, K-ATPase, acetylcholinesterase and 5'-nucleotidase were about 5, 6-fold, 2, 5-fold and 3, 3-fold, respectively, enriched in the plasma membrane fraction as compared to those of the crude homogenate. Electron microscpic examination also showed that the membranes were in vesicular form.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2001년도 제18차 정기총회 및 학술발표 PROCEEDINGS
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• pp.71-73
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• 2001

본 연구는 원시생식기 유래 원시생식세포를 이용하여 효율적인 생식선 카이메라 생산 조건을 확립하고자 하였다 전체 실험구에서 301마리가 부화하였으며, 이중 수컷 141마리, 암컷 160마리가 성성숙에 도달하였다. 후대 검정을 통하여 수컷 15마리와 암컷 12마리가 생식선 카이메라로 밝혀졌으며, 생식선 카이메라 생산 효율은 평균 9.0%였다. 실험처리구간 생식선 카이메라 생산효율간에 유의적인 차이는 없었으나 (p=0.6831), 생식선 카이메라의 공여체 유래 자손 생산 효율은 유의적인 차이를 나타냈다. 실험처리구간 생식선 카이메라의 공여체 유래 자손 생산효율은 피콜처리 없이 10일간 배양한 원시생식세포를 이식하였을 때 가장 높았고 (49.7%), 피콜처리후 배양하지 않은 원시생식세포를 이식하였을 때 가장 낮았다 (0.6%). 또한, 피콜 처리에 상관없이 10일간 배양한 원시생식세포를 이식한 실험구가 배양하지 않은 실험구와 5일간 배양한 실험구보다 각각 50배와 10배 더 높은 전이효율을 나타내었다 (p<0.0001). 따라서, 수용체 배자에 이식하기전에 실시한 체외배양 기간은 효율적인 생식선 카이메라 생산에 있어서 매우 중요한 요인으로 사료된다. 이와 같이, 본 연구에서는 배양 기간, 피콜 처리 유무 등에 대한 비교 분석을 통하여 생식선 카이메라 생산을 위한 최적 조건을 확립하였다.

• Journal of Plant Biology
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• 제33권4호
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• pp.325-332
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• 1990

Light membrane vesicles were isolated from the zucchini hypocotyl by floatation on ficoll density gradients and the proteins were solubilized with Triton X100. Three ATP-hydrolyzing enzymes were partially purified by ion-exchange and gel filtration chromatography and isoelectric focusing. There are plasma membrane-type ATPase whose activity was inhibited by vanadate but not by nitrate, tonoplast-type ATPase which was sensitive to nitrate but insensitive to vanadate and one having a phosphatase activity with a pI value different from that of an acid phosphatase. A fraction was obtained after DEAE-ion-exchange chromatography crossreacting with polyclonal antibodies against Ca2+ -ATPase from human erythrocytes.