• Title/Summary/Keyword: Primary culture

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Effects of ${\beta}$-Mercaptoethanol on the Growth of Preantral Follicles and the Maturation of Intrafollicular Oocytes

  • Gong, Seung Pyo;Lim, Jeong Mook
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.35-41
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    • 2009
  • This study was undertaken to evaluate how ${\beta}$-mercaptoethanol (bME), an exogenous antioxidant, interacts with preantral follicles cultured in vitro. Mouse primary or secondary follicles were cultured in glutathione (GSH)-free or GSH-containing medium supplemented with bME of various concentrations, and the growth of preantral follicles, the maturation of intrafollicular oocytes and preimplantation development after parthenogenesis were monitored. In experiment 1, 0, 25, 50 or 100 ${\mu}M$ bME was added to culture medium supplemented with 100 ${\mu}M$ GSH or not. When secondary follicles were cultured in GSH-free medium, no significant change in follicle growth was detected after bME addition. However, exposure to bME in the presence of GSH significantly inhibited both follicle growth and oocyte maturation. Such detrimental effect became prominent in primary follicles and bME strongly inhibited follicle growth in the absence of GSH. In conclusion, there are stage-dependent effects of bME on follicle growth and oocyte maturation, and selective use of antioxidants contributes to establishing an efficient follicle culture system.

Effect of Extracellular Calcium on Vitellogenin Production in the Culture of Hepatocytes in the Rainbow Trout, Oncorhynchus mykiss

  • Yeo In-Kyu;Mugiya Yasuo;Chang Young Jin;Hur Sung Bum;Yoo Sung Kyu
    • Fisheries and Aquatic Sciences
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    • v.1 no.1
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    • pp.24-29
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    • 1998
  • Effect of extracellular calcium in vitellogenin (VTG) production in response to estradiol-17 $\beta$ $(E_2,\;2\times10^{-6}M)$ was examined in primary hepatocyte culture of rainbow trout, Onchorhynchus mykiss. Total calcium in estrogenized sera significantly increased, compared with the control, while diffusible calcium was insignificant. However, diffusible calcium in the incubation medium with $E_2$ was significantly reduced, compared with the control. The uptake of extracellular calcium by cultured hepatocytes signifIcantly increased 90 min after $E_2$ addition. Moreover, the accumulation of intracellular calcium increased in the cultures with $E_2$, regardless of the calcium concentrations in the incubation media. In addition, $E_2-primed $ VTG production was significantly decreased by withdrawal of E_2$ from the incubation medium. Moreover, VTG production by $E_2-primed$ hepatocytes was reduced by removing calcium from the incubation medium with or without $E_2$. These results suggest that the entry of extracellular calcium into the cytoplasm is an important step for VTG production in primary hepatocyte cultures in rainbow trout.

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

  • Hwang, Un-Gl;Park, Jin-Il;Shim, Jung-Min;Jung, Chang-Soo;Park, Sung-Yoon
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2003.11a
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    • pp.159-164
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    • 2003
  • Effects of Al on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 $\beta$($E_2$) were examined in primary hepatocyte culture of rainbow trout. Hepatocytes were precultured for 2 days and then E2 ($2{\times}10^{-6}$M) and Al ($10^{-6}-10^{-4}$M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metal had no appreciable effect on the viability of hepatocytes in culture. However, Al interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction accurred at Al concentrations greater than $5{\times}10^{-5}$M. VTG mRNA expression also decreased with a negative correlation with Al concentration (r=-0.98). These results suggest that Al inhibit VTG production at the transcriptional level to reduce VTG mRNA expression.

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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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New Perspectives in Pottery Typology of Korean Archaeology - Related to the Typology of Chungdo-Type Pottery from the Youngdong Region - (토기의 형식분류론에서 제기되는 몇 가지 문제에 대하여 - 영동지역 출토「중도식」토기편년과 관련하여 -)

  • Lee, Jun-Ho
    • Korean Journal of Heritage: History & Science
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    • v.36
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    • pp.87-104
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    • 2003
  • The Chungdo-type Pottery Culture, distributed through the middle part of the Korean peninsula, is chronologically located in the very former stage of the advent of ancient states. It has two different traditions of pottery manufacturing technique which are totally different in choosing raw materials, shaping, fixing and firing. It seems that two different traditions had been selectively applied by pottery type. In order to understand this peculiar cultural aspect, the pottery typology needs to be different from those applied to cultures where pottery was made and used under the single manufacturing tradition. This study tries to find the new pottery typology which best fits for the understanding the chronology of the Chungdo-Type Pottery Culture. For this purpose, I examined existing typologies, recognized their problems, and then build a new typology. As a result, I found that the former typologies misinterpreted the relative frequencies of each pottery type as different function or region. In this article, I propose the new pottery typology as building a primary classification within each function and region, and then synthesizing all of primary classifications. This new typology eliminates the factors of function and region in understanding the chronology of the Chungdo-Type Pottery Culture, and assorts the regional distinction by comparing pottery types in each region.

Effect of Culture Conditions on Cathepsin B Inhibitor Production by a Marine Bacterium, Pseudomonas sp. Strain PB01

  • Hoang, Le Thu Van;Kim, Moon-Moo;Kim, Se-Kwon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1115-1120
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    • 2008
  • A novel cathepsin B inhibitor-producing bacterium was isolated from marine sediments and identified based on its 16S rDNA sequence as Pseudomonas sp. strain PB01 (Accession No. EU126129). The growth and enzyme inhibitor production were investigated under various culture conditions. A mixture of organic nitrogen source was required for the optimal production, whereas both glucose and maltose proved to be the effective carbon sources for cathepsin B inhibitor production. Other optimal culture conditions included temperature range between 25 and $28^{\circ}C$, initial medium pH of 6.6, and shaking speed of 200 rpm. Under these optimal conditions, the maximum inhibitory activity from culture broth was approximately 50% after 30 h of cultivation. Additionally, kinetic study revealed that inhibitor production paralleled with cell growth, which suggested that the inhibitor may be a primary metabolite of that bacterium.

A Digital Thesaurus of the Traditional Common Culture of the Greater Mekong Subregion

  • Suwannee Hoaihongthong;Kanyarat Kwiecien
    • Journal of Information Science Theory and Practice
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    • v.12 no.3
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    • pp.63-74
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    • 2024
  • This study aimed to develop a digital thesaurus dedicated to cataloging the traditional common culture of the Greater Mekong Subregion. The process followed a meticulous seven-step methodology, including scoping, vocabulary collection, knowledge structure analysis, relationship delineation, related word adjustments, list validation, and evaluation. Leveraging principles from knowledge organization, thesaurus construction, and digital platform development, the TemaTres web application emerged as the primary tool for constructing this thesaurus. The study's results showed that 2,042 principal words related to the traditional common culture of the Greater Mekong Subregion were compiled and classified into terms for each of the seven deep levels. Each term was accompanied by essential metadata, including broader and narrower terms, related terms, cross-references, and scope notes. This rich dataset empowered semantic search capabilities across diverse applications and web services, providing access to knowledge pertaining to the traditional common culture of the Greater Mekong Subregion and contributing to a deeper understanding of this cultural domain.

Characterization of a Cell Line HFH-T2, Producing Viral Particles, from Primary Human Fetal Hepatocytes Infected with Hepatitis B Virus

  • Shim, Jae-Kyoung;Kim, Dong-Wook;Chung, Tae-Ho;Kim, June-Ki;Suh, Jeong-Ill;Park, Chun;Lee, Young-Choon;Chung, Tae-Wha;Song, Eun-Young;Kim, Cheorl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.186-192
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    • 2001
  • A primary culture of human fetal hepatocytes was obtained through a therapeutic abortion process at 26 weeks of gestation period. More than $10^8$ cells were seeded on a plastic plate. These hepatocytes were infected with hepatitis B virus (HBV). The HBV was purified from serum of one chronic HBV carrier. Transformed hepatocytes were subcultured in a 10% FBS-supplemented medium. The morphology of the transformed cell was epithelial-like. The cells from the first pass showed signs of early proliferation and had a latent period of more than 3 months after 6-7 passages. After the rest period, the transformed cell proliferated actively and they were subcultured every three days. Transformed hepatocytes were characterized by detection of the HBV transcript by RT-PCR. The secretion of virions from transformed cells was investigated by PCR with the cell medium. Two types of virions secreted into the culture medium were examined by using the transmission electron microscope. Another approach to study the secretion of virions in to culture medium was carried out with HBV antibody. HBsAg was detected in the culture medium of transformed cells using ELISA and Western blot analyses. These data suggested that the human fetal hepatocyte cell line has been established by infection of HBV, in which this cell line secreted viral particles into the culture medium.

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The Investigation of Cell Culture Conditions to Maintain Chicken Embryonic Stem Cells as Totipotent Cells

  • Du, Lixin;An, Jing
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.8
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    • pp.1102-1107
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    • 2003
  • The ES cell can provide a useful system for studying differentiation and development in vitro and a powerful tool for producing transgenic animalds. To investigate the culture condition of chicken embryonic stem (CES) cells which can retain their multipotentiality or totipotency, three kinds of feeder layer cells, SNL cells, primary mice embryonic fibroblasts (PMEF) cells and primary chicken embryonic fibroblasts (PCEF) cells, were used as the feeder cells in media of DMEM supplemented with leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and stem cell factor (SCF) for co-culture with blastoderm cells from stage X embryos of chicken. The alkaline phosphatase (AKP) test, differentiation experiment in vitro and chimeric chicken production were carried out. The results showed that culture on feeder layer of PMEF yielded high quality CES cell colonies. The typical CES cells clone shape revealed as follows: nested aggregation (clone) with clear edge and round surface as well as close arrangement within the clone. Strong alkaline phosphatase (AKP) reactive cells were observed in the fourth passage cells. On the other hand, the fourth passage CES cells could differentiate into various cells in the absence of feeder layer cells and LIF in vitro. The third and fourth passage cells were injected into the subgerminal cavity of recipient embryos at stage X. Of 269 Hailan embryos injected with CES cells of Shouguang Chickens, 8.2% (22/269) survived to hatching, 5 feather chimeras had been produced. This suggests that an effective culture system established in this study can promote the growth of CES cells and maintain them in the state of undifferentiated and development, which lays a solid foundation for the application of CES cells and may provide an alternative tool for genetic modification of chickens.