• Title/Summary/Keyword: high density cell culture

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식물세포의 관류배양을 위한 초음파 분리기의 개발

  • Gu, Yeong-Han;Jo, Gyu-Heon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.402-404
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    • 2000
  • In this study, we have developed an ultrasonic separation system for plant cells and its operating conditions in terms of voltage, flow rate and concentration were examined. For plant cell, the operation of ultrasonic separator highly depended on concentration of cells. Holding capacity highly depended on flow rate in chamber. Optimum voltage was 30V in high density culture

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High Cell Density Culture of Alcaligenes eutrophus and Poly-$\beta$-hydroxybutyrate Production by Optimization of Medium Compositions (배지조성 최적화를 통한 Alcaligenes eutrophus의 고농동 세포배양 및 Poly$\beta$-hydroxybutyrate 생산)

  • 이용우;유영제
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.401-406
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    • 1994
  • The medium compositions of Alcaligenes eutrophus were optimized for increasing PHB productivity. It is very important to optimize the concentrations of inorganic salts and trace eleme- nts as well as carbon and nitrogen sources to maximize cell growth rate and productivity. The fed-batch culture of Alcaligenes eutrophus by dual feeding of ammonia water and glucose under optimized initial medium concentrations was carried out. Glucose was fed manually according to glucose consumption rate and ammonia water by pH-stat. The final cell concentrations and PHB content in 30 hours were 122 g/l and 65% of dry cell weight(yielding 79 g of PHB/l), respectively and 2.64 g/l/hr of PHB production rate was obtained.

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세라믹 필터를 장착한 생물반응기에서 Bacillus thuringiensis의 성장 특성 모델링

  • Gang, Byeong-Cheol;Jang, Ho-Nam
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.233-236
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    • 2000
  • Bacillus thuringiensis (Bt) is the most widely used microbial insecticide in the biological control market. Cultivation of the microorganism to high cell densities offers potential for enhancing the rate of formation as well as the concentration of the desired products In the fermentation broths in bioreactor. With this objective, we developed the new bioreactor incorporating ceramic membrane module for the retention of cell mass. Cell yield and spore formation of Bacillus thuringiensis was improved markedly by adopting this new bioreactor based on glucose -limited feeding operation. It was possible to grow the cell and the heat-resistant spore to above $1.2\;{\times}\;10^{10}\;CFU/ml$ density. With glucose-limited operation, we studied the growth behavior of Bacillus thuringiensis during the cell retention culture. Linear growth of Bacillus thuringiensis was observed under glucose-limited culture, which matched well with simple mathematical model of cell retention culture.

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Metabolic Analysis of Poly(3-Hydroxybutyrate) Production by Recombinant Escherichia coli

  • WONG, HENG HO;RICHARD J. VAN WEGEN;JONG-IL CHOI;SANG YUP LEE
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.593-603
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    • 1999
  • Poly(3-hydroxybutyrate) (PHB) production by fermentation was examined under both restricted- and ample-oxygen supply conditions in a single fed-batch fermentation. Recombinant Escherichia coli transformed with the PHB production plasmid pSYLl07 was grown to reach high cell density (227 g/l dry cell weight) with a high PHB content (78% of dry cell weight), using a glucose-based minimal medium. A simple flux model containing 12 fluxes was developed and applied to the fermentation data. A superior closure (95%) of the carbon mass balance was achieved. When the data were put into use, the results demonstrated a surprisingly large excretion of formate and lactate. Even though periods of severe oxygen limitation coincided with rapid acetate and lactate excretion, PHB productivity and carbon utilization efficiency were not significantly impaired. These results are very positive in reducing oxygen demand in an industrial PHA fermentation without sacrificing its PHA productivity, thereby reducing overall production costs.

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Selection of Protoplasts-Derived Cell Lines for High Yields Indole Alkaloids from Suspension Cultures of Vinca (Catharanthus roseus) (일일초(Catharanthus roseus) 현탁배양으로부터 원형질체유래 Indole Alkaloid 고생산성 세포주 선발)

  • 김석원;정경희
    • KSBB Journal
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    • v.6 no.1
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    • pp.1-7
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    • 1991
  • To produce economically important indole alkaloids by cell cultures, we selected protoplastsderived clones (protoclones) of vinca (Catharanthus roseus) for high yields of catharanthine and ajmalicine. Protoplasts were enzymatically isolated from suspension-cultured cells. The highest plating efficiency (1%) was obtained when protoplasts were plated at a density of 1$\times$105 protoplasts/ml in a culture medium solidified with 0.4% Seaplaque agarose. The growth rates of 40 protoclones subcultured on a solid medium varied over a wide range. Protoclone VPC-6, which had the highest growth rate, was observed to produce relatively high yields of catharanthine and ajmalicine when cultured in a liquid medium. Although the original cell line did not produce catharanthine at a detectable level by HPLC, protoclone VPC-10 produced it at a level of 5.9$\mu\textrm{g}$/g fresh weight of cells for 10 days of culture. Under the same conditions, protoclone VPC-15 produced ajmalicine at a level of 133.6$\mu\textrm{g}$/g, of which productivity was improved about ,3 times than that of the original cell line. The results indicate that differences in the growth rate and indole alkaloid yield among the protoclones reflect the somaclonal variation in suspnsion-cultured cells.

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Production of Extracellular Polysaccharide by Perfusion Culture of Angelica gigas Nakai Suspension Cells (배지교환식 고농도 배양에 의한 참당귀 현탁세포 유래 ECP 생산)

  • Kim, Young-Hwa;Kim, Ik-Hwan;Kim, Dong-Il
    • KSBB Journal
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    • v.21 no.5
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    • pp.336-340
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    • 2006
  • High-density perfusion cultivation was performed to produce extracellular polysaccharide(ECP) as immunostimulating agents in suspension cell cultures of Angelica gigas Nakai. In batch culture, the maximum cell density was 16.8 gDCW/L at day 6 and 0.9 g/L of ECP was obtained at day 8. When the medium exchange was started at the fifth day after inoculation for the perfusion culture, high concentration of the cells at 23.8 gDCW/L could be achieved with continuous production of ECP. Treatments of ultrasound and Pluronic F-68 were found to be helpful for the secretion of intracellular ECP into the culture medium.

Curcuma Longa L. Extract Controls Cancer Cell (Sarcoma 180) Growth (강황(薑黃) 추출물의 암세포 성장 억제 효과)

  • Cho, Su-In
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.27-31
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    • 2005
  • Objectives : The anticancer response of three different types of water extracts of Zingiberaceae Curcuma longa L. tested for sarcoma 180. Only few studies carried out to investigate the effects of other contents of Curcuma longa L. in anticancer activities, therefore, in this study we have investigated the effects of other component then curcumin in Curcuma longa L. for anticancer a activities. Methods : Three different types of water extracts of Curcuma longa L. were prepared as follows. The sarcoma cells (S180) were maintained in Dulbecco's modified Eagle's medium (DMEM) and were seeded on 24-well cell culture cluster flat bottom with lid tissue culture treated non-pyrogenic polystyrene. The growth of sarcoma 180 was monitored for 1, 2 and 5 days. The sarcoma cells were pictured using inverted microscope and cell density was counted using hemocytometry. Results : After 5 days in the culture medium the results showed high growth of sarcoma 180 for control condition and the surface of CCP plates were fully covered with the cells. In case of medium in which the 10% of filtered water extract of Curcuma longa L. was added a very limited growth of sarcoma 180 was observed. The results were showed only small difference in cell density for two different concentrations of unfiltered water extracts of Curcuma longa L. whereasin case of filtered water extracts the control of sarcoma growth shows better result. Conclusion : The filtered water extracts showed the best result relatively to the unfiltered water extracts for two different concentrations. This indicates that the water extracts of Curcuma longa L. can have anticancer activities possibly without curcumin.

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Production of tissue-type plasminogen activator from immobilized CHO cells introduced hypoxia response element

  • Bae, Geun-Won;Kim, Hong-Jin;Kim, Gi-Tae;Kim, Ik-Yeong
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.257-260
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    • 2002
  • Dissolved oxygen level of cell culture media has a critical effect on cellular metabolism, which governs specific productivity of recombinant proteins and mammalian cell growth However, in the cores of cell aggregates or cell-immobilized beads, oxygen level frequently goes below a critical level. Mammalian cells have a number of genes induced in the lower level of oxygen, and the genes contain a common cis-acting element (-RCGTG-), hypoxia response element (HRE). By binding of hypoxia inducible factor-l (HIF-I) to the HRE, promoters of hypoxia inducible genes are activated, which is a survival mechanism. In this work, to develop a CHO cell capable of producing recombinant proteins in immobilization and high density cell culture efficiently, mammalian expression vectors containing human tissue-type plasminogen activator (t-PA) gene controlled by HRE were constructed and stably transfected into the CHO cells. In $Ba^{2+}$ -alginate immobilization culture, CHO/pCl/dhfr/2HRE-t-PA cells produced 2 folds higher recombinant t-PA activity than CHO/pCl/dhfrlt-PA cells without $CoCl_2$ treatment. Furthermore, in repeated fed batch culture, productivity of t-PA in immobilized CHO/pCI/dhfr/2HRE-t-PA cells was 121 ng/ml/day, total production of 0.968 mg/day at 11 days culture while CHO/pCIIdhfrlt-PA cells was 22.8 ng/ml/day. All these results indicate that HRE is very useful for the enhancement of protein productivity in mammalian cell cultures.

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