• Title/Summary/Keyword: Mammalian cell

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A Novel Helicosporium Isolate and Its Antimicrobial and Cytotoxic Pigment

  • Choi, Hye Jung;Lee, Sang Myeong;Kim, Sun-Hee;Kim, Dong Wan;Choi, Young Whan;Joo, Woo Hong
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1214-1217
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    • 2012
  • One Helicosporium strain, isolated from a wilted chestnut tree, evidenced in vitro antimicrobial activity against various types of bacteria and fungi, and generated a diffusible pigment. The antimicrobial compounds and the diffusible pigment of the Helicosporium sp. isolate were purified via solvent fractionation, column chromatography, and recycling preparative chromatography. Both the major antimicrobial compound and the diffusible pigment were identified as 2-methylresorcinol via nuclear magnetic resonance spectroscopy. Therefore, 2-methylresorcinol, a diffusible pigment generated by Helicosporium sp., appears to be an active antimicrobial principle. This pigment also exhibited considerable cytotoxicity against mammalian cells.

Small GTPases and formins in mammalian oocyte maturation: cytoskeletal organizers

  • Kwon, So-Jung;Lim, Hyun-Jung J.
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.1
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    • pp.1-5
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    • 2011
  • The maturation process of mammalian oocytes accompanies an extensive rearrangement of the cytoskeleton and associated proteins. As this process requires a delicate interplay between the cytoskeleton and its regulators, it is often targeted by various external and internal adversaries that affect the congression and/or segregation of chromosomes. Asymmetric cell division in oocytes also requires specific regulators of the cytoskeleton, including formin-2 and small GTPases. Recent literature providing clues regarding how actin filaments and microtubules interact during spindle migration in mouse oocytes are highlighted in this review.

Ion Transports in Mouse Collecting Duct Cells (M-1)

  • Park, Mi-Young;Song, Jin-Mi;Jang, Phil-Geum;Lee, Ryang-Hwa;Jung, Jin-Sup
    • Proceedings of the Korean Biophysical Society Conference
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    • 1999.06a
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    • pp.50-50
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    • 1999
  • The mammalian cortical collecting duct (CCD) plays a major role in regulating renal NaCl absorption, which is important in controlling total body Na and Cl homeostasis. The M-1 cell line, derived from the mouse cortical collecting duct, is being used as a mammalian model of the CCD to study electrolytes transport.(omitted)

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Genotoxicity Study of sophoricoside in bacterial and mammalian cells system

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.177.1-177.1
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    • 2003
  • Sophoricoside was isolated as the inhibitor of IL -5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develope as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test and mouse lymphoma tk gene assay (MOLY). (omitted)

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Genotoxicity Study of sophoricoside derivatives in mammalian cells system

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.178.2-178.2
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    • 2003
  • To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of Quantity. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay(MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. (omitted)

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Recent Advanced Toxicological Methods for Environmental Hazardous Chemicals (환경 오염물질의 진보된 독성 평가 기법)

  • 류재천;최윤정;김연정;김형태;방형애;송윤선
    • Environmental Analysis Health and Toxicology
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    • v.14 no.1_2
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    • pp.1-12
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    • 1999
  • Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk$\^$+/-/ gene assay (MOLY) using L5178Y tk$\^$+/-/ mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk$\^$+/-/ gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells, Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D. C. in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

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High Productivity of t-PA in CHO Cells Using Hypoxia Response Element

  • Bae Gun-Won;Jeong Dae-Won;Kim Hong-Jin;Lee Gyun-Min;Park Hong-Woo;Choe Tae-Boo;Kang Seong-Man;Kim Ick-Young;Kim Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.695-703
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    • 2006
  • The dissolved oxygen level of any cell culture environment has a critical effect on cellular metabolism. Specifically, hypoxia condition decreases cell viability and recombinant protein productivity. In this work, to develop CHO cells producing recombinant protein with high productivity, mammalian expression vectors containing a human tissue-type plasminogen activator (t-PA) gene with hypoxia response element (HRE) were constructed and stably transfected into CHO cells. CHO/2HRE-t-PA cells produced 2-folds higher recombinant t-PA production than CHO/t-PA cells in a $Ba^{2+}-alginate$ immobilized culture, and 16.8-folds in a repeated batch culture. In a non-aerated batch culture of suspension-adapted cells, t-PA productivity of CHO/2HRE/t-PA cells was 4.2-folds higher than that of CHO/t-PA cells. Our results indicate that HRE is a useful tool for the enhancement of protein productivity in mammalian cell cultures.

Effect of Dietary Fat on Structure and Function of Mammalian Cell Membrane (식이지방이 생체막 구조와 기능에 미치는 영향)

  • Cho, Sung-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.4
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    • pp.459-468
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    • 1984
  • The currently accepted model of membrane structure proposes a dynamic, asymmetric lipid matrix of phospholipids and cholesterol with globular proteins embedded across the membrane to various degrees. Most phospholipids are in the bilayer arrangement and also closely associated with integral membrane proteins or loosely associated with peripheral proteins. Biological functions of membrane, such as membrane-bound enzyme functions and transport systems, are influenced by the membrane physical properties, which are determined by fatty acid composition of phospholipids, polar head group composition and membrane cholesterol content. Polar and non-polar region of the phospholipid molecule can interact, with changes in the conformation of a membrane-associated protein altering either its catalytic activity or the protein's interaction with other membrane proteins. Mammalian dietary studies attempted to change the lipid composition of a few cell membranes have shown comparisons, using essential fatty acid-deficient diets. In recent years, Clandinin and a few other workers have pioneered the study proving the influence of dietary fat fed in a nutritionally complete diet on composition of phospholipid classes of cell membrane. Modulation caused by diet fat was rapid and reversible in phospholipid fatty acyl composition of membranes of cardiac mitochondria, liver cell, brain synaptosome and lymphocytes. These changes were at the same time, accompanied by variety of membrane associated functions controlled by membrane-bound enzymes, tranporter and receptor proteins. The findings suggest the basic concept of the necessity of dietary fatty acid balance if consistency of optimal membrane structural lipid composition is to be maintained, as well as the overall inadequacy of describing the nutritional-biochemical quality of a dietary fat solely by its content of linoleic acid. Furthermore, they give light on the possible application to clinical and preventive medicine.

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Investigating the Morphology and Kinetics of Three-Dimensional Neuronal Networks on Electro-Spun Microstructured Scaffolds

  • Kim, Dongyoon;Kim, Seong-Min;Kang, Donghee;Baek, Goeun;Yoon, Myung-Han
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.277.2-277.2
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    • 2013
  • Petri dishes and glass slides have been widely used as general substrates for in vitro mammalian cell cultures due to their culture viability, optical transparency, experimental convenience, and relatively low cost. Despite the aforementioned benefit, however, the flat two-dimensional substrates exhibit limited capability in terms of realistically mimicking cellular polarization, intercellular interaction, and differentiation in the non-physiological culture environment. Here, we report a protocol of culturing embryonic rat hippocampal neurons on the electro-spun polymeric network and the results from examination of neuronal cell behavior and network formation on this culture platform. A combinatorial method of laser-scanning confocal fluorescence microscopy and live-cell imaging technique was employed to track axonal outgrowth and synaptic connectivity of the neuronal cells deposited on this model culture environment. The present microfiber-based scaffold supports the prolonged viability of three-dimensionally-formed neuronal networks and their microscopic geometric parameters (i.e., microfiber diameter) strongly influence the axonal outgrowth and synaptic connection pattern. These results implies that electro-spun fiber scaffolds with fine control over surface chemistry and nano/microscopic geometry may be used as an economic and general platform for three-dimensional mammalian culture systems, particularly, neuronal lineage and other network forming cell lines.

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Development of High Density Mammalian CellCulture system for the Production of Tissue-Type Plasminogen Activator

  • Park, Byong-Gon;Chun, Joo-Mi;Lee, Chang-Jin;Chun, Gie-Taek;Kim, Ik-Hwan;Jeong, Yeon-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.123-129
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    • 2000
  • A high cell density culture system for the anchorage dependent CHO cells was developed based on the combination of in removal of ammonium ion and microcarrier culture system, and semi-fed-batch feeding of glucose and glutamine was employed to the developed culture system. The glass bead was selected as an optimum microcarrier in terms of cell growth. An ammonium ion selective zeolite, Phillipsite-Gismondine, was packed in a dialysis menium ion. The semi-fed-batch operation was employer to the novel culture system for the high density cell culture, and the results showed the cell growth was improved by 32% and tPA productivity by 250%.

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