• Title/Summary/Keyword: 초대배양

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Vitellogenin Induction by Rainbow trout (Oncorhynchus mykiss) Hepatocytes in Primary Culture (무지개송어의 간세포 초대배양에 의한 Vitellogeinin 합성 유도)

  • 여인규
    • Journal of Aquaculture
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    • v.11 no.4
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    • pp.557-564
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    • 1998
  • Vitellogenin (VTG) induction in response estradiol-17${\beta}$ ($E_2$) were electrophoretically examined in primary hepatocyte cultures in rainbow trout. The hepatocytes were maintained as monolayers on positively charged dishes for up to 7 days. The viability of hepatocytes on Day 7 in cultures decreased about 20.7% and 23.6% with and without $E_2$, respecitively. The amount of DNA per dish also decreased to 13.7% and 14.0% with and without $E_2$, respectively. There were no differences in viability and DNA content between the control and $E_2$-treated culture. Moreover, the rate of VTG to total protein concentrations reached the maxium level at the addition of $10^{-6}$ M E2, to the incubation medium. However, the higher concentration of $10^{-5}$ M $E_2$ rater depressed the level.

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감성돔(Acanthopagrus schlegeli)에서 유래한 새로운 세포주의 확립

  • 임은영;강민수;오명주;정성주
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2003.05a
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    • pp.318-319
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    • 2003
  • 어류에서 바이러스성 질병이 중요하다는 인식이 증가하고 있으며 어류바이러스의 연구를 위한 세포주의 발달이 활성화되고 있다. 하지만상대적으로 해산유래의 세포 주 수는 많지 않다. 특히 유럽과 우리나라 남ㆍ서해안에서 고급어종인 감성돔은 림포 시스티스병, 이리도바이러스병 등의 바이러스성 질병이 발생하고 있다. 본 연구는 감성돔에서 유래한 세포주 개발을 목적으로 하였으며, 감성돔의 비장에서 유래한 세포를 초대배양 및 계대배양 하여 새로운 세포주를 확립하였으며, 일련의 실험을 실시하여 확립된 세포주의 특성을 확인하고 바이러스에 대한 감수성을 검사하였다. (중략)

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In vitro Mass Propagation of Ardisia pusilla DC. (산호수 (Ardisia pusilla DC.)의 기내 대량번식)

  • Kang Gwan-Ho;Oh Owel-Sun;Goo Dae-Hoe;Eun Jong-Seon;Kim Hyung-Moo
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.281-285
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    • 2005
  • To establish the mass proliferation system of Ardisia pusilla DC, the shoot tips of Ardisia pusilla DC were cultured on the MS and half-strength MS medium supplemented with $0{\sim}5.0$ mg/L BA or $0{\sim}0.5$ mg/L thidiazuron(TDZ), respectively. A few multiple shoot formation observed when the shoots were cultured on MS medium containing TDZ. However, the frequency of multiple shoot formation was reached up to 82.4%, when the shoots were cultured on half-strength MS medium supplemented with 0.5 mg/L BA. Also the number of shoot per explant was 7.1. To promote rooting from multiple shoot, newly formed shoots were transferred to half-strength MS medium containing 0.5 mg/L IBA or 0.5 mg/L NAA, respectively. Regenerated plantlets were grown to normal mature plants in soil.

Screening of Hepatoprotective Substances from Higher Fungi by Primary Cultured Rat Hepatocytes intoxicated with Carbon Tetrachloride ($CCl_4$로 독성을 유발시킨 초대배양 간세포를 이용한 고등균류로부터 간세포 보호물질의 검색)

  • Lee, June-Woo;Han, Man-Deuk;Lee, Kweon-Haeng
    • The Korean Journal of Mycology
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    • v.20 no.4
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    • pp.347-353
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    • 1992
  • Hepatoprotective effects of polysaccharides extracted from liquid cultured mycelia were screened by measuring the glutamic pyruvate transaminase activity of the primary cultured rat hepatocytes intoxicated with carbon tetrachloride. Sixty among 75 isolates of higher fungi showed to be hepatoprotective effect, and these were 13 of Ganoderma lucidum, 5 of Lentinus edodes, 1 of Pleurotus ostereatus, 4 of Coriolus versicolor, 2 of Lyophyllum spp., 7 of Grifora frondosa, 3 of Agaricus spp., 14 of Schizophyllum commune and 11 of Cordyceps spp.. Especially, 10 isolates, Ganoderma lucidum IY003 and IY009, Lentinus edodes IY103, Lyophyllum sp. IY402, Agaricus sp. IY701 and IY703, Schizophyllum commune IY804, IY810 and IY818, Cordyceps sp. IY902, were indicated below 80% of glutamic pyruvate transaminase activity.

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Efficient Bulblet Regeneration and Growth from Bulb Scale of Hyacinthus orientalis L. cv. Pink Pearl Cultured in vitro (히아신스(cv. Pink Pearl)의 인편 기내 배양시 효과적인 자구의 재생과 생장)

  • Lee, Kyung-Soon;Chung, Chung-Han;Chung, Yong-Mo;Kim, Doh-Hoon;Jeong, Soon-Jae;Nam, Jae-Sung;Kim, Gyung-Tae;Yi, Young-Byung
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1336-1340
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    • 2007
  • The regeneration and growth of bulblets from the bulb scale segments of Hyacinthus orientalis L. cv. Pink Pearl were more efficient in IBA than IAA at the same concentrations (1.0 and 3.0 mg/l). The normal (base-down) orientation of explants was more effective for bulblet regeneration and root growth than the inverted (base-up) orientation. The growth of bulblets and roots was increased higher in the perlite than the agar medium. These results suggested that the alternate culture system, first cultured in the agar medium for bulblet regeneration, and then in the perlite medium for bulblet growth, may be more useful for efficient in vitro culture of hyacinth (H. orientalils) cv. Pink Pearl.

Effects of insulin and IGF on growth and functional differentiation in primary cultured rabbit kidney proximal tubule cells - Effects of IGF-I on Na+ uptake - (초대배양된 토끼 신장 근위세뇨관세포의 성장과 기능분화에 대한 insulin과 IGF의 효과 - Na+ uptake에 대한 IGF-I의 효과 -)

  • Han, Ho-jae;Park, Kwon-moo;Lee, Jang-hern;Yang, IL-suk
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.783-794
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    • 1996
  • It has been suggested that ion transport systems are intimately involved in mediating the effects of growth regulatory factors on the growth of a number of different types of animal cells in vivo. The functional importance of the apical membrane $Na^+/H^+$ antiporter in the renal proximal tubule is evidenced by estimates that this transporter mediates the reabsorption of approximately one third of the filtered load of sodium and the bulk of the secretion of hydrogen ions. This study was designed to investigate the pathway utilized by IGF-I in regulating sodium transport in primary cultured renal proximal tubule cells. Results were as follows : 1. $Na^+$ was observed to accumulate in the primary cells as a function of time. Raising the concentration of extracellular NaCl induced an decrease in $Na^+$ uptake compared with control cells in a dose dependent manner. The rate of $Na^+$ uptake into the primary cells was about two times higher in the absence of NaCl($40.11{\pm}1.76pmole\;Na^+/mg\;protein/min$) than in the presence of 140mM NaCl($17.82{\pm}0.94pmole\;Na^+/mg\;protein/min$) at the 30 minute uptake. 2. $Na^+$ uptake was inhibited by IAA($1{\times}10^{-4}M$) or valinomycin($5{\times}10^{-6}M$) treatment($50.51{\pm}4.04$ and $57.65{\pm}2.27$ of that of control, respectively). $Na^+$ uptake by the primary proximal tubule cells was significantly increased by ouabain($5{\times}10^{-5}M$) treatment($140.23{\pm}3.37%$ of that of control). When actinomycin D($1{\times}10^{-7}M$) or cycloheximide($4{\times}10^{-5}M$) was applied, $Na^+$ uptake was decreased to $90.21{\pm}2.39%$ or $89.64{\pm}3.69%$ of control in IGF-I($1{\times}10^{-5}M$) treated cells, respectively. 3. Extracellular cAMP decreased $Na^+$ uptake in a dose-dependent manner($10^{-8}-10^{-4}M$). IBMX($5{\times}10^{-5}M$) also inhibited $Na^+$ uptake. Treatment of cells with pertussis toxin(50pg/ml) or cholera toxin($1{\mu}g/ml$) inhibited $Na^+$ uptake. Extracellular PMA decreased $Na^+$ uptake in a dose-dependent manner(1-100ng/ml). 100 ng/ml PMA concentration significantly inhibited $Na^+$ uptake in IGF-I treated cells. However, staurosporine($1{\times}10^{-7}M$) had no effect on $Na^+$ uptake. When PMA and staurosporine were added together, the inhibition of $Na^+$ uptake was not observed. In conclusion, sodium uptake in primary cultured rabbit renal proximal tubule cells was dependent on membrane potentials and intracellular energy levels. IGF-I stimulates sodium uptake through mechanisms that involve some degree of de novo protein and/or RNA synthesis, and cAMP and/or PKC pathway mediating the action mechanisms of IGF-I.

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Effect of L-Ascorbic Acid on Collagen Synthesis in 3T6 Fibroblasts and Primary Cultured Cells of Chondrocytes (3T6 세포주 및 연골 초대배양세포의 Collagen 합성에 미치는 비타민 C의 영향)

  • Kim, Mi-Hyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.42-47
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    • 2006
  • L-Ascorbic acid (AsA) is an essential nutrient for prevention of scurvy in humans, primates and guinea pigs that lack $L-gulono-\gamma-lactone$ oxidase which is required for the final step of AsA biosynthesis. AsA participates in various hydroxylation reactions involved in the biosynthesis of collagen. The purpose of this study is to clarify the role of AsA on collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. Cells were cultured in medium supplemented with catalase and AsA at various concentration. Supplement of AsA induced collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. The most remarkable induction of collagen synthesis by AsA was found in primary cultured chondrocytes. The content of collagen representing the amounts of extracellular matrix significantly increased in the cells of which growth was stimulated by AsA, while it decreased with increasing passage numbers of subculture in cells. It showed that the content of collagen decreased in the medium which contained AsA at the concentration higher than 5.0 mM. However, the contents of collagen to DNA were not different among various AsA concentrations. Supplementing with AsA resulted in enhancement of collagen formation and extracellular matrix. Therefore, there might be a Positive correlation between the activity of catalase and the AsA concentration. Moreover, it can be assumed that AsA stimulates the collagen synthesis by optimizing the cell-culture environment.

Effects of Bisphenol A on Vitellogenin Synthesis in the Hepatocyte Primary Culture in the Olive Flounder, Paralichthys olivaceus (넙치 Paralichthys olivaceus 초대 배양 간세포의 Vitellogenin합성에 미치는 Bisphenol A의 영향)

  • Yeo, In-Kyu;Choe, Mi-Kyung
    • Korean Journal of Ichthyology
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    • v.12 no.3
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    • pp.180-185
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    • 2000
  • The effects of bisphenol-A(BPA), a monomer of plastics used in many consumer products, on vitellogenin(VTG) synthesis were examined in primary hepatocyte culture of olive flounder, Paralichthys olivaceus. The hepatocytes were precultured for 2 days, and then estradiol-$17\beta(10^{-6}M)$ and BPA were simultaneously added to the incubation medium. The hepatocytes were cultured for 6 more days and then spent medium was analyzed by SDS-PAGE. BPA increased the rate of VTG to total protein concentrations in a concentration-dependent way, and a significant difference was obtained at concentrations of $10^{-6}M$ and $10^{-5}M$ (P<0.05). In particular, the rate of VTG to total protein concentrations was 26.36% at $10^{-5}M$ of BPA, and its level did not differ from the control level with $E_2$ alone. $E_2$ and/or BPA-primed VTG synthesis was markedly inhibited to about 80% of the control(with $E_2$) by the addition of tamoxifen($10^{-6}M$) to the incubation medium. Furthermore, In vivo $E_2$-primed VTG synthesis was significantly inhibited by in vitro $E_2$-free incubation of hepatocyte to about 22% of the control (with $E_2$) on Day 6. The effect of reducing was delayed in a BPA concentration-dependent way. These results suggest that BPA induce VTG synthesis by estrogenic activity through estrogen receptor mediated response and prolong VTG synthesis on vitellogenesis in olive flounder.

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