• 제목/요약/키워드: Primary Culture

검색결과 1,126건 처리시간 0.031초

간세포 배양-약물대사를 위한 모델 연구 (I. Primary cultured hepatocytes as a key in vitro model to improve preclinical drug development)

  • 이경태
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 제2회 추계심포지움
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    • pp.135-140
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    • 1994
  • Over past decades, numerous in vitro model has been developed to investigate drug metabolism. In the order of complexity we found the isolated perfused liver, hepatocytes in co-culture with epithelial cells, hepatocytes in suspension and in primary culture and subcellular hepatic microsomal fractions. Because they can be easily prepared from both animals (pharmacological and toxicological species) and humans (whole livers as well as biopsies obtained during surgery) hepatocytes in primary culture provide the most powerful model to better elucidate drug behavior at an early stage of preclinical development such as : 1. the characterization of main biotransformation reactions. 2. the identification of phase I and phase II isozymes involved in such reactions 3. the evaluation of interspecies differences allowing the selection of a second toxicological animal species more closely related to man on the basis of metabolic profiles 4. the detection of the inducing and/or inhibitory effects of a drug on metabolic enzymes, the prediction of drug interactions 5. the estimation of inter-individual variability in biotransformation reactions. The use of hepatocytes, and in particular those obstained from humans, at an early stage of drug development allows the obtention of more predictive preclinical data and a better knowledge of drug behavior in humans before the first administration of the drug in healthy volunteers.

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전통색에 관한 연구 (A Study on Traditional Colors)

  • 최인려;방혜경;김여주
    • 복식문화연구
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    • 제16권2호
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    • pp.397-407
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    • 2008
  • Recently, influence of color has been expanded in various fields including design field. Among human five senses, visual sense is best for information capturing, and 80% of information judgment by visual sense is from color. Color is emotion and image. Hanbok is marked by its beautiful color combinations. The guiding principle of such color chords was Obangsaek, or the traditional Korean five base colors. The method of this study is to consider concepts of Obangsaek derived from the principles of yin-yang and the five elements and is to investigate application of Obangsaek. And the purpose of this study is to understand traditional colors and is to provide judgment criteria on various color combinations based on Korean aesthetic sense for development of textile design. The degree of nation culture depends on the level of research, analysis and application on traditional colors. Obangsaek, Korean traditional color is splendid and primary color. Also, Obangsaek has symbolic and lucky meanings. These colors are still the primary source of coloring. Obangsaek has been reinterpreted by application on textile design, some into base colors and others into diverse shades. So this study will help in the aspect of development on Korean style design.

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콩 유묘의 조직배양에 의한 식물체 분화 (Plant Regeneration from in vitro Tissue Culture of Soybean Seedling)

  • 김용호;김석동;홍은희
    • 한국작물학회지
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    • 제37권5호
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    • pp.419-424
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    • 1992
  • 콩에서의 조직배양 가능성을 알아보고자 콩 유묘를 기내에서 배양시킨 뒤, 여러 부위를 채취하여 각기 다른 배지 치상한 결과는 다음과 같다. 1. 줄기정단조직 배양에서는 대부분 shoot가 분화되어 완전한 식물체를 얻을 수 있었다. 2. 자엽절 조직배양에서는 multiple shoots가 나타나 대량증식에 유용했으나 배지에 따른 영향이 컸다. 3. 배축 및 상배축 조직배양에서는 callus만 분화되엇다. 4. 엽조직배양에서는 callus 분화 후 계대배양했을 때 shoot가 분화되었으나 완전한 식물체를 얻기는 힘들었다.

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Vitellogenin and Its mRNA Induction by $Estradiol-17\beta$ in the Primary Culture of Hepatocytes in the Rainbow Trout, Oncorhynchus mykiss

  • Hwang Un-Gi
    • Fisheries and Aquatic Sciences
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    • 제4권4호
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    • pp.186-191
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    • 2001
  • Vitellogenin (VTG) and VTG mRNA induction by $estradiol-17\beta\;(E_2)$ were examined in the primary cultures of hepatocyte in the rainbow trout. Hepatocytes were precultured for 2 days, then $E_2$ was added and cultured for another 5 days. Media and hepatocytes were then analyzed by electrophoresis and Northern blotting for VTG and VTG mRNA, respectively. The hepatocytes were formed a few aggregates within 5 days without further spreading to a monolayer. Cell viability and high DNA content were maintained during the incubation. The hepatocyte culture with E2 induced a weak VTG band at a molecular weight of 175kDa on Day 2 after $E_2$ addition. The relative amount of VTG was expressed in percentage of total protein concentrations. VTG was gradually increased as $1.9\%$ on Day 2, $6.3\%$ on Day 4 and $7.3\%$ on Day 5. VTG mRNA band was detected at about 6.6 kb in the culture with $E_2$ at day 1 of culture. The level of VTG mRNA expression linearly increased with time until Day 5 (r=0.97).

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CYTOTOXICITY OF D-GALACTOSAMINE ON PRIMARY CULTURES OF ADULT RAT HEPATOCYTES

  • Yang, K.H.;Park, Kwan-Ha;Kim, Byung-Sam
    • Toxicological Research
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    • 제3권2호
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    • pp.73-80
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    • 1987
  • Primary cultures of adult rat hepatocytes were used to study the cytotoxicity of D-galactosamine. Hepatocytes were isolated by a collagenase perfusion technique and maintained as monolayers in serum-free medium on collagen-coated culture dishes. Treatment of galactosamine to the culture markedly inhibited the uptake of ${\alpha}$-aminoisobutyric acid (AIB) inducible with glucagon and dexamethasone. At0.1 mM of galactosamine, AIB uptake was inhibited significantly when treated for 12 hr. At higher doses (0.25, 0.5 and 1.0mM), a significant inhibition was noticed after 1 hr exposure. Generally the magnitude of the inhibition was related to the dose and treatment time of galactosamine. Treatment of galactosamine also produced a dose- and treatment time-related suppression of the tyrosine aminotransferase (TAT) induction caused by dexamethasone. Meanwhile, uptake of ouabain was not affected by the treatment of galactosamine. The viability of the hepatocytes was decreased only slightly by the treatment of galactosamine; more than 87% of the cells excluded tryphane blue when treated 1 mM galactosamine for 12 hr. Galactosamine induced depressions of AIB uptake and TAT activity were prevented by the simultaneous addition of uridine to the culture. D-Galactosamine, cytotoxicity, hepatocytes culture, ${\alpha}$-aminoisobutyric acid uptake, tyrosine aminotransferase.

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내분비 장애물질 검출을 위한 In Vitro Bioassay 개발 : 어류 혈청을 이용한 간세포 단층배양 (Development of In Vitro Bioassay for Detection of Estrogenic Activity of Xenobiotics : Monolayer Culture of Hepatocytes using Fish Serum)

  • 권혁추;맹준호;김은희;최성희
    • 한국발생생물학회지:발생과생식
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    • 제13권4호
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    • pp.217-226
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    • 2009
  • 본 연구는 내분비 장애물질의 검출을 위하여 간세포의 단층 형성, 생존 및 기능에 미치는 어류 혈청의 영향에 대해 검토하였다. 한국산 메기의 간세포는 자신의 혈청 및 뱀장어, 틸라피아 등 타어종의 혈청에 의해 부착 및 단층이 형성되었으나, FBS는 메기 간세포의 단층을 형성시키지 못했다. 0.5에서 3%의 어류 혈청으로 메기 간세포의 단층을 형성 시킬 수 있는데, 이것은 FBS(5~20%) 사용의 1/10 이하로 적은 양이며, 어류 혈청이 FBS를 대체할 수 있고, FBS보다 간세포의 형태 및 기능 유지에 효과적인 것으로 나타났다. 어류 혈청이 첨가된 배양액에서 메기 간세포는 적어도 10일 이상 단층 형성을 유지할 수 있어, 내분비 장애물질 연구에 이용될 수 있을 것이다. 결론적으로 본 연구에서 개발된 어류 혈청을 사용한 메기 간세포 배양시스템과 효소면역측정법(ELISA)은 bisphenol A 등의 내분비 장애물질의 검출 및 연구를 위한 유용한 도구로서 이용될 수 있다고 생각된다.

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DETECTION OF DNA SINGLE-STRAND BREAKS AND UNSCHEDULED DNA SYNTHESIS INDUCED BY PROCARCINOGENS IN PRIMARY CULTURES OF RAT HEPATOCYTES

  • Kim, D.H.;Kim, Bok-Ryang;K. H. Yang
    • Toxicological Research
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    • 제2권1호
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    • pp.1-7
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    • 1986
  • Procarcinogen induced DNA single-strand breaks and unschduled DNA synthesis were measured in primary rat hepatocytes culture. For DNA single-strand breaks assay, rat liver DNA was prelabeled by injection 3H-thymidine during the peak of DNA synthesis following partial hepatectomy. Hepatocytes were isolated from the rat 2 weeks after surgery by a collagenase perfusion techinique and maintained as monolayers in serum free medium on collagen-coated culture dishes. DNA sigle-strand breaks were measured by the alkaline elution techinique.

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EFFECT OF DITHIOL MALONATE DERIVATIVES (DMDs) ON CARBON TETRACHLORIDE-INDUCED HEPATOTOXICITY IN PRIMARY CULTURES OF ADULT RAT HEPATOCYTES

  • Jung, Hyun-Ho;Jeong, Tae-Cheon;Yang, Kyu-Hwan;Chun, Young-Jin
    • Toxicological Research
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    • 제9권2호
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    • pp.167-175
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    • 1993
  • Protective effects of dithiol malonate derivatives (DMDs), YH-100, YH-150 and YH-439 on carbon tetrachloride-induced hepatotoxicity were investigated in primary rat hepatocytes culture. Treatment of DMDs to hepatocytes culture did not affect total cytochrome P-450 content and ECOD and AHH activities. Protein and RNA synthesis was also similar to control. Meanwhile, DMDs significantly decreased LDH release and in vitro lipid peroxidation induced by $CCI_4$. Accumulation of cellular triglyceride and decreased secretion of VLDL from liver cells by $CCI_4$ treatment were also significantly protected.

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계대세포를 이용한 소 수정란의 체외배양 체계에 관한 연구 (Effects of established cell lines on bovine embryo development during in vitro culture)

  • 주석천;이병천;이원유;최윤석;황우석
    • 대한수의학회지
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    • 제37권3호
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    • pp.647-659
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    • 1997
  • To overcome the difficulties of collecting and culture of primary cell from genital tract on embryonic development, the present study was carried out to investigate the critical effect of cell lines, such as BRL and Vero cell and its conditioned medium on the development of early Korean native cattle embryos in vitro. Oocytes collected from slaughterhouse ovaries were matured in TCM199 containing FSH, estradiol-$17{\beta}$ and FBS with granulosa cell monolayer for 24 hours and then fertilized in vitro using frozen-thawed, heparin-treated spermatozoa in TALP for 30 hours. And then early embryos (1-2cell) were cultured in TCM199 containing 10% FBS with BOEC, Granulosa, BRL, Vera cell monolayers and conditioned medium for 2~3 days. Development to morulae and blastocysts were recorded, also examined the number of blastomeres presented a valuable parameter for the evaluation of embryonic development. The early cleavage rates of in vitro fertilized embryos co-cultured, there was no differences between primary cell and cell lines(p<0.05). The rate of development to the later stage, coculture of BRL cell was significantly higher than that of the primary cell(p<0.05). The rates of development to morula and blastocyst were significantly higher in vero cell than BRL, Granulosa, Oviduct epithelial cell conditioned medium. In the result of effect of serum on development of early bovine embryos, the use of media containing serum were significantly higher than the use of not containing one on development of early and later stage of embryos. The result of number of blastomeres in blastocysts, there is no differences between primary cell and cell lines. The blastocysts from coculture were higher than from conditioned medium in blastomere cells. In summary, these experments have proved that the culture system in TCM199 with BRL, Vero cell monolayers is effective on in vitro development of early bovine embryos, In addition, it is effective to development of bovine embryos that containing serum in conditioned medium, or in co-culture rather than in conditioned medium alone. The use of cell lines opponent to primary cells is effective in bovine embryo culture.

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분리 세포의 단층세포 배양법에 의한 인체 비점막 상피세포의 배양 (In Vitro Culture of Human Nasal Epithelial Cells by Monolayer Culture of Dissociated Cells)

  • 김용대;송시연;민명기;서장수;송계원;박호선
    • Journal of Yeungnam Medical Science
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    • 제15권2호
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    • pp.286-296
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    • 1998
  • 비중격 만곡증 환자의 비내수술시 채취한 정상 하비갑개 점막조직으로 부터 상피세포만을 분리세포의 단층배양법(monolayer culture of dissociated cells)으로 배양하여 비점막 상피세포 배양법을 정립하고 또한 배양된 세포가 상피세포임을 동정하기 위하여 간접 면역형광항체법으로 상피세포 특유의 cytokeratin을 확인하였고 투과전자현미경으로 상피세포만이 가지고 있는 교소체(desmosome) 및 장세사(tonofilament) 등을 확인하였으며 6회까지 계대배양을 할 수 있었다. 향후 본 연구에서 배양된 비점막 상피세포를 이용하여 알레르기성 비염 등과 같은 비점막 염증성 질환의 병태생리에서 상피세포의 역할에 대한 연구와 호산구등 염증 세포의 침윤과 상피세포의 손상에 접착분자 및 cytokine의 상호작용에 관한 연구가 계속될 수 있을 것으로 기대한다.

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