• Title/Summary/Keyword: Cell cultures

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Establishment of Cell Suspension Cultures and Plant Regeneration in White Dandelion (Taraxacum coreanum NAKAI.)

  • Sun, Yan-Lin;Kim, Jae-Hak;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.24 no.3
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    • pp.280-285
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    • 2011
  • In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

Antioxidant Effect of Poncirin and Cytotoxicity on Cultured Human Skin Fibroblast Damaged by Methyl Mercury

  • Jung, In-Ju;Back, Jong-Cheon;Choi, Yu-Sun
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.355-360
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    • 2007
  • In order to evaluate on the cytotoxicity of methyl mercury (MM) and antioxidant effect of phenolic compound, poncirin against MM-induced cytotoxicity, XTT assay was performed to determine the cell viability after human skin fibroblasts (Detroit 51) were grown in the media containing various concentrations of methylmercuric chloride (MMC). And also, the antioxidant effect of poncirin on the cytotoxicity induced by MMC was examined by cell viability and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in these cultures. MMC decreased cell viability in dose-dependent manner in these cultures and the midcytotoxicity value was determined at concentration of 30 ${\mu}M$ MMC after human skin fibroblasts were treated with $10\sim50{\mu}M$ MMC for 72 hours, respectively. MMC was highly toxic on cultured human skin fibroblasts by toxic criteria. MMC-mediated cytotoxicity was related with oxidative stress by the diminution of toxic effect according to the treatment of vitamin E. In the antioxidant effect of poncirin, it showed vitamin E-like DPPH radical scavenging activity at 90 ${\mu}g/ml$ poncirin and also, remarkably increased cell viability compared with MMC-treated group. From these results, it is suggested that MMC-mediated cytoxicity was highly toxic and was related with oxidative stress in cultured human skin fibroblasts, and also phenolic compound such as poncirin showed the protection on MMC-induced cytotoxicity by antioxidant effect in these cultures.

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Development of a Magnetic-field Stimulation System for Cell Cultures in situ: Simulation by Finite Element Analysis

  • Dominguez, G.;Arias, S.;Reyes, Jose L.;Rogeli, Pablo
    • Journal of Magnetics
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    • v.22 no.2
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    • pp.326-332
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    • 2017
  • The effects of exposure to an extremely low-frequency magnetic field (25 Hz 20G) on animal cells have been studied. In some reports, stimulation was performed for fixed frequency and variations in magnitude; however, animal-cell experiments have established that both parameters play an important role. The present work undertook the modeling, simulation, and development of a uniform-magnetic-field generation system with variable frequency and stimulation intensity (0-60 Hz, 1-25G) for experimentation with cell cultures in situ. The results showed a coefficient of variation less than 1 % of the magnetic-field dispersion at the working volume, which is consistent with the corresponding simulation results demonstrating a uniform magnetic field. On the other hand, long-term tests during the characterization process indicated that increments of only $0.4^{\circ}C$ in the working volume temperature will not be an interfering factor when experiments are carried out in in situ cell cultures.

Increased lignan biosynthesis in the suspension cultures of Linum album by fungal extracts

  • Bahabadi, Sedigheh Esmaeilzadeh;Sharifi, Mozafar;Safaie, Naser;Murata, Jun;Yamagaki, Tohru;Satake, Honoo
    • Plant Biotechnology Reports
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    • v.5 no.4
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    • pp.367-373
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    • 2011
  • Linum album accumulates anti-tumor podophyllotoxin (PTOX) and its related lignans, which were originally isolated from an endangered species Podophyllum. In the present study, we examined the effects of five fungal extracts on the production of lignans in L. album cell cultures. Fusarium graminearum extract induced the highest increase of PTOX [$143{\mu}g\;g^{-1}$ dry weight (DW) of the L. album cell culture], while Rhizopus stolonifer extract enhanced the accumulation of lariciresinol up to $364{\mu}g\;g^{-1}$ DW, instead of PTOX. Typical elicitors, such as chitin, chitosan, or methyl jasmonate (MeJA), were shown to be less effective in lignan production in L. album cell cultures. These results verified the advantages of fungal extracts to increase lignan production in L. album cell culture, and suggested potential on-demand metabolic engineering of lignan biosynthesis using differential fungal extracts.

Selection of Herbicide Tolerant Cell lines from $\gamma$-ray-Irradiated Cell Cultures in Rice (Oryza sativa L. cv. Ilpumbyeo)

  • Bae, Chang-Hyu;Lee, Young-Ill;Lim, Yong-Pyo;Seo, Yong-Won;Lee, Do-Jin;Yang, Deuk-Chun;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.4 no.3
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    • pp.123-127
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    • 2002
  • Herbicide tolerant rice (Oryza sativa L. cv. Ilpumbyeo) cell lines were selected from $\gamma$-ray-irradiated anther-derived cell cultures. The anther-derived cell clusters were small (300 to 400 ${\mu}{\textrm}{m}$ in diameter) and uniform ones that were screened by miracloth filtering. The cell suspensions were very efficient to plate one layer onto agar medium and to screen target cell lines. Herbicide tolerant cell lines were selected by 5 mg/L cyhalofop butyl (CHB) treatment by using the small cell suspensions on agar N6 medium containing 1 mg/L 2,4-D and 0.2 mg/L kinetin. Of the cell lines, one line (CHB-1) showed stable tolerance at 10 mg/L concentration after 6-month culture without herbicide suspension. Growth stability of CHB-1 was similar to that of control cell line on 10 mg/L CHB containing medium. In this experiment we established herbicide tolerant cell line selection system by using anther-derived uniform-cell suspensions with $\gamma$-ray-irradiation.

Effects of Initial Inoculum Size, Liquid Volume and Medium Feeding Strategy on Panax ginseng Hairy Roots Growth

  • Jeong, Gwi-Taek;Park, Don-Hui;Hwang, Baek
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.250-253
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    • 2000
  • We researched effects of growth on initial inoculum size, liquid volume, and medium feeding rate etc. Cell suspension inoculated at low cell concentrations showed a typical growth reduction, whereas root cultures showed an improvement in growth. In this paper, Hairy roots showed high growth rate at 0.4 % inoculum size and 100 mL liquid volume in 250 mL flask cultures.

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Bioprocess Considerations for Production of Secondary Metabolites by Plant Cell Suspension Cultures

  • Chattopadhyay, Saurabh;Farkya, Sunita;Srivastava, Ashok K.;Bisaria, Virendra
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.3
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    • pp.138-149
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    • 2002
  • Plant cell culture provides a viable alternative over whole plant cultivation for the production of secondary metabolites. In order to successfully cultivate the plant cells at large scale, several engineering parameters such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for selection of a suitable bioreactor. The media ingredients, their concentrations and the environmental factors are optimized for maximal synthesis of a desired metabolite. Increased productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular metabolites. Proper understanding and rigorous analysis of these parameters would pave the way towards the successful commercialization of plant cell bioprocesses.

Effects of Phosphate, Precursor and Exogenous Berberine on the Production of Alkaloid in Plant Cell Cultures

  • Kim, Dong-Il
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.79-83
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    • 1991
  • The effects of phosphate concentration in the medium, feeding of biosynthetic precursor, and the addition of exogenous berberine on cell growth and berberine production were studied in cell suspension cultures of Thalictrum rugosum. The depletion of phosphate in the medium enhanced the specific productivity up to twofold with significant release of berberine into the medium. Extracellular berberine was 19% of the total in the culture without phosphate while it was 2-5% of total berberine in the culture with even low amounts of phosphate. Precursor feeding was not effective in enhancing alkaloid formation. Initial presence of exogenous berberine did not have much effect on cell growth and alkaloid production. It was found that the cells have the capacity to take up large quantities of berberine. When $500{\;}mg{\cdot}l^{-1}$ of berberine was added exogenously at the beginning, 81% of total berberine was found in the cells.

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Effects of Stireed Tank Bioreactor Scale-up on Cell Growth and Alkaloids Production in Cell Cultures of Eschscholtzia californica (탱크 교반형 생물반응기의 scale-up이 Eschscholtzia californica 세포생장 및 알칼로이드 생성에 미치는 영향)

  • 유병삼;변상요
    • KSBB Journal
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    • v.13 no.6
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    • pp.700-705
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    • 1998
  • Studies were made to investigate effects of the scale-up of stirred tank bioreactors on cell growth and alkaloids production for suspension cultures of Eschscholtzia californica. In the 1.5 L STR, cell lysis was observed at 110 rpm or higher agitation speed. The agitation speed of 30 L STR was 43.7 rpm to maintain the same shear stress developed in 1.5 L STR of 100 rpm. As a result of scale-up from 1.5 L to 30 L STR, the specific growth rate was decreased from 0.12 to 0.07 day-1. The alkaloids productivity was also decreased from 0.24 to 0.14 mg/L-day. Changes of mixing performance and oxygen transfer were studied to explain the decrease of cell growth and alkaloids production. Decreased oxygen transfer rate coefficient(KLa) and increased mixing time by the scale-up was observed at various aeration rates.

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Effects of Nutrients and Culture Conditions on the Cell Growth and the Flavonol Glycosides Production in Cell Cultures of Ginkgo biloba (Ginkgo biloba 세포배양에서 배지 및 배양조건이 세포성장 및 Flavonol Glycosides 생합성에 미치는 영향)

  • 이원규;유연우변상요정헌관
    • KSBB Journal
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    • v.8 no.1
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    • pp.55-61
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    • 1993
  • Cell lines of Ginkgo biloba were derived from different plant parts and from ten varieties spanning various geographic locations. They had various properties of growth and product formation. More than three flavonol glycosides were present in low concentration in callus and suspension cultures. Cell growth and biosynthesis of flavonol glycosides were found to be affected by medium composition. Culture conditions which influenced cell growth and product formation were also examined. Light stimulated the flavonol glycosides biosynthesis and ten times higher flavonol glycosides content was obtained as compared with the result without light.

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