• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.313-316
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• 2002

Effect of Pluronic F-68, a nonionic surfactant, on the extracellular production of hGM-CSF in transgenic Nicotiαna tabacum cell suspension culture was investigated. The addition of 5 g/L Pluronic F-68 did not affect the cell growth but increased the extracellular production of hGM-CSF by two-fold. This may be due to the enhanced permeability of the cell membrane by the interaction between the Pluronic F-68 and the cell membrane.

• The Korean Journal of Zoology
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• v.30 no.4
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• pp.401-409
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• 1987

단백질 분해효소와 ovoinhibitor가 계배근세포 융합에 미치는 영향을 알아보기 위하여 세포 운합지수의 측정, Western blot 분석법에 의한 ovoinhibitor의 세포 내농도 측정 및 단백질 분해효소의 활성도를 조사하였다. 세포 배양 30시간과 72시간에 처리된 ovoinhibitor는 상당히 근세포 융합 억제시켰다. 근세포 내 ovoinhibitor의 농도는 배 양 48시간에서 가장 높았고, 배양이 진행됨에 따라 계속 떨어져서 대체로 융합지수가 올라갈수록 그 농도가 낮아지는 경향을 보였다. 근세포 분화가 진행됨에 따라 총 단백질 분해효소의 환성도는 계속 증가하였는데 특히 배양 48시간에서 72시간 사이의 급격한 증가를 나타내는 시기는 세포 융합이 가장 활발히 일어나는 시기와 일치했다. 이상의 결과는 단백질 분해효소 저해제가 근세포 분화에 상당한 영향을 끼침을 보여 주며, 단백질 분해효소와 단백질 분해효소 저해제는 하나의 조절단위로써 근세포 분화에 어떤 작용을 할 것으로 사료된다.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.87-87
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• 2001

도살장에서 구입한 소의 난소로부터 회수한 미성숙 난포란을 체외에서 성숙, 수정시킨 후, 2~8세포기 수정란을 CR$_1$aa 체외배양액에 일정량의 aesculetin, taurine을 첨가하여 체외배양을 실시한 두 처리구간의 항산화 효과를 비교 검토하고, aesculetin과 taurine에 growth factor(EGF, PDGF)를 첨가배양하여 체외수정란의 체외발육에 항산화제와 growth factor의 상호작용 효과를 검토하였다. 대조구, aesculetin(1$\mu\textrm{g}$/$m\ell$) 및 taurine(2.5mM)을 첨가하여 체외수정란을 체외배양시킨 결과 상실배이상 발육된 체외발육율은 각각 46.5%, 64.3% 및 60.5%로서 대조구보다 aesculetin과 taurine 첨가구가 유의하에 높은 성적을 얻어, aesculetin과 taurine이 체외수정란의 체외발육에 높은 효과를 나타냈다. 대조구, aesculetin (1$\mu\textrm{g}$/$m\ell$) ＋ PDGF (1ng/$m\ell$) 첨가구, aesculetin (l$\mu\textrm{g}$/$m\ell$) ＋ EGF (10ng/$m\ell$)첨가구, taurine (2.5mM) ＋PDGF (1ng/$m\ell$) 첨가구 및 taurine (2.5mM)＋EGF (10ng/$m\ell$) 첨가구에서 배반포기 발육율은 각각 46.0%, 70.0%, 58.0%, 56.0 및 66.0%로써 처리구가 무첨가구에 비해 유의하게 높은 성적을 얻어(P<0.05), 항산화제와 growth factor의 첨가 배양이 체외수정란의 체외발육에 상승효과가 있음이 입증되었다. 배양액에 천연추출 aesculetin과 상품화된 aesculetin을 첨가하여 체외배양을 실시한 결과 배반포기 발육율은 대조구, 천연추출 aesculetin 및 상품화된 aesculetin 첨가구에서 각각 38.6%, 54.6% 및 53.5%로써 첨가구가 무첨가구보다 높은 성적을 얻어, aesculetin의 항산화 효과가 입증되었다(P<0.05). 그러나 모든 처리구에서 배반포까지 발육된 체외수정란의 세포수에는 커다란 차이가 인정되지 않았다.

• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.77-84
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• 1996

The purpose of this study was to evaluate effects of osmolarity and vitamins on the in vitro development of embryos. Bovine embryo that had been matured and fertilized in vitro were cultured in simple, chemically defined, protein free medium (mTLP-PVA). When the osmolarity of the medium supplemented 0.35 mM phos-phate and 19 amino acid was changed by NaCI concentration, significantly(p<0.05) higher pro-portion of 1-cell embryos in the medium of 265 or 290 mOsm developed to the morula (32 ~ 35%) and blastocyst(24 ~ 28%) stage. When embryos were transferred to fresh medium containing 5.56mM glucose at 120hrs post-insemination, the highest proportion of embryos developed to mor-ula(40%) and blastocyst(32%) stages at 290 mOsm(p<0.05), although the value in morulae was not significantly different with that(35%) at 315 mOsm. Vitamins in presence of glutamine and amino acids had no beneficial effects on the development of 1-cell embryos to the blastocyst.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.614-618
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• 1992

The batch and fed-batch cultivations of Brevibacteriurn CHI were carried out for the production of nitrile hydratase. In batch culture, with pH control the cell mass and the specific activity increased more 20% and 30%. respectively. The maximum growth rate was obtained at a glucose concentration of $20g/{\ell}$ because of substrate inhibition. The fed-batch culture of Brevibacteriurn CHI with constant substrate feeding gave a cell density of up to $68g/{\ell}$ and nitrile hydratase activity was maintained at above 6.1units/mg. The cell growth yield on carbon .source was ca. 0,68 g/g glucose consumed. The total nitrile hydratase activity in this fed-batch mode increased up to 414.8 units/m${\ell}$, which amounted to 4.4 times that of the batch culture.

• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.33-36
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• 2001

The experiments were conducted to establish the in vitro culture system of apple rootstock M.9. The meristem tissue of M.9 were pre-treated in antiox: dant solution containing 100 mgL$^{-1}$ ascorbic acid and 150 mgL$^{-1}$ citric acid for 30 minutes, transferred to the MS liquid medium added with 0.1 mgL$^{-1}$ IBA, 0.5 mgL$^{-1}$ GA, and 30 gL$^{-1}$ sucrose, which shaked by 50 rpm for 2 weeks, and then, cultured in same composition of MS agar medium. This treatment stimulated shooting from the tissue, the most favorably, compared with other treatments. All young shoots produced normal roots when they were shake-cultured on the 1/2MS liquid medium added with 0.5 mgL$^{-1}$ IBA, 30 gL$^{-1}$ sucrose and 1,000 times diluted solution of Hormex by 50 rpm for one week, and subsequently transferred to the 8 gL$^{-1}$ agar medium of the same composition as pre-culture medium minus Hormex.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.272-280
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• 1991

The effect of media feeding rate on cell growth and monoclonal antibody production in the fed-batch culture ot hybridoma A4W was studied. In the batch culture, the highest specific antibody production rate was observed at the begining of the culture period but its value tended to decrease rapidly with the culture time. The final antibody concentration and volumetric productivity was 65 $\mu g$/ml and 13 mg Mab/l/day, respectively. In the fed-batch culture, the specific antibody production rate, $q_p$ rebounded sharply within a few hours after the media feeding was started and it remained high until the end of culture if the media feeding was continued. The final antibody concentration was 220 $\mu g$/ml and the volumetric productivity was 45.1 mg/l/day. Further increase in final antibody concentration was achieved by applying a modified media of which component was fortified with glucose and glutamine, hence the final antibody concentration in this case was 270 $\mu g$/ml and the volumetric productivity was 51.8 mg/lday, which is as four tinlcs as high cuixparinf! to that of batch culture.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.325-333
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• 1997

수정란이식의 주변기술인 초기배의 미세조작및 성 판정은 가축의 경우, 경제 형질의 유전적 개량에 크게 기여하였다. 본 연구는 미세조작 biopsy와 PCR에 의한 체외생산 소 초기배의 급속. 정확한 성 판정 기법을 확립하기 위해 실시하였다. 체외성숙 및 체외수정에 의해 생산된 소수정란은 소 난관상피세포와 공배양을 통해 8-세포부터 배반포시시까지 체외발생시킨 후 미세조작 biopsy에 이용되었다. 미세조작 biopsy 과정중에 약간의 형태적인 손상이 관찰되었지만 대부분의 demi-embryo는 정상적인 배반포와 나화배반포로 발생하였다. 8~16 세포, 상실배, 초기배반, 포 시기에서 미세조작 biopsy 후 확장 또는 나환배반포기시기까지의 발달율은 각각 62.8(27/43), 83.3(30/36) 및 80.9%(55/68)로 정상적인 초기배와 큰 차이가 없는 것으로 나타났다. 총 136개의 소 초기배로 부터 2~10개의 할구세포를 미세조작 biopsy에 의해 분리하여 소 특이와 Y-특이염기서열을 가진 두 쌍의 염기서열을 이용하여 PCR을 수해하였다. 이들중 112(82.4%)개를 성공적으로 성 판정하였으며, 암/수 비율은 각각 34.8(39/112)/65.2%(73/112)로 나타났다. 본 실험에서 얻은 결과는 확립된 소 초기배의 미세조작기술과 성 판정 방법을 통해 계획적인 암.수 송아지의 선별 생산뿐만 아니라 소 수정란 이식을 통한 유전적 개량을 촉진시키는데 효과적으로 이용될 수 있는 기술이 확립된 것을 보여주었다.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.97-108
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• 2000

The objectives of this study were performed to increase the efficiency of the culture conditions of embryos produced in vitro, and to assess the developmental potential after transfer of those embryos into recipients. The mean number of folliclular oocytes recovered from an ovary was 10.7. The rates of maturation and fertilization in Grade I oocytes were significantly (P＜0.05) higher than Graden and III. Developmental rate into blastocyst in the culture group of TCM-199 with BOEC were significantly higher (P＜0.05) than the groups of TCM-199 and conditioned medium (24.7% vs. 12.4% and 18.2%). The survivability of post-thawed blastocysts equilibrated for 3 min in EFS solution was significantly (P＜0.05) lower than l0 for 1 and 2 min (32.1% vs. 82.9% and 73.3%). Significantly higher (P＜0.05) survival rate in blastocysts was seen after freezing than in morulae stage embryos. Out of all 105 recipients, 49 (46.7%) were confirmed in pregnant. On pregnancy of cattle, 48 calves were born from 40 recipients. The ratio of twin and single calves was 30.5% (32/40 and 7.6% (8/40), respectively. However, the others composed of abnormal, as judging as 6 (12.2%) for abortion and 3 (6.1%) for stillbirth during the pregnant period.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.69-74
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• 1998

The somatic embryos of Daucus carota L. cv. Hongshim developed in MS basal medium far 4 weeks had varied number of cotyledons. Palisade and spongy parenchyma of cotyledon were not clearly discriminated in all seedlings developed from the embryos. No independent existence of collateral vascular bundle was observed in all seedlings with various types of cotyledon ; instead, vascular bundles were either interconnected or partially connected with one another. Most of the cotyledonary bases on hypocotyl showed short cylinder structure which encircle plumule. The vascular tissues of cotyledonary bases and nodes of seedlings with jar-shaped or 1 cotyledon were connected in ring forms, showing the pattern of ectophloic shiphonostele, and similar ring form structure was also found in the vascular arrangement of 5 cotyledon seedlings. The vascular bundles of seedlings with 2, 3 and 4 cotyledons in many cases had independently arranged within the cotyledonary bases and nodes, showing the pattern of eustele. In hypocotyl, tetrarch or hexarch xylems prevailed in seedlings with jar-shaped cotyledon or 1 and 5 cotyledon; tetrarch xylems prevailed in 2 cotyledon seedlings; and triarch xylems prevailed in 3 cotyledon seedlings. In most of seedlings, cortex vascular bundles were dispensed in the region from cotyledonary node to hypocotyl, but double vascular bundles were also observed occasionally. In roots, diarch xylems were observed in most of seedlings with 2 cotyledons, triarch xylems in half of seedlings with 3 cotyledons, and diarch xylems in most of the remaining seedlings with the occasional occurrences of tetrarch xylems.