• Title/Summary/Keyword: high density cell culture

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An Experimental Study on Bacterial Adhesion onto Activated Carbon and Ceramic (활성탄 및 세라믹 재질에의 세균 부착성에 대한 연구)

  • Kwon Sung-Hyun;Cho Dae-Chul;Rhee In-Hyoung
    • Journal of Environmental Science International
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    • v.14 no.12
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    • pp.1163-1170
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    • 2005
  • The microbial adsorption characteristics of two different media for biological treatment were studied using attached diverse microbes onto activated carbon and ceramic. The results in the experiments of the characteristics of physical adhesion on two different media with addition of high and low concentrated substrate in the culture were observed that the efficient of adhesion onto F-400 activated carbon was higher over that of ceramic due to the surface area of media. The irradiation treatment by ultrasonication with 400 W power and 3 min retention time on the media without addition substrate conditions and subsequent mixing throughly the culture showed the highest efficiency of cell detachment on the media. Three different microbes, P. ovalis, A calcoaceticus, and B. subtillis were used for the study of the characteristics of microbial adhesion on the media. p ovalis showed the highest adhesion capability while B. subtillis showed the lowest capability adhesion onto media either addition of substrate in the culture. The mixed bacterial culture showed $10\%$ lower removal efficiency of DOC in the low concentrated substrate culture compared to the single pure culture. Whileas, it did not show significant difference between two cultures at high concentrated substrate. It was also observed same population density of microorganism by counting of microbes adhered to microbial media with an ultrasound treatment.

Effects of Environmental Factors on Cyanobacterial Production of Odorous Compounds: Geosmin and 2-Methylisoborneol

  • Oh, Hyung-Seok;Lee, Chang Soo;Srivastava, Ankita;Oh, Hee-Mock;Ahn, Chi-Yong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1316-1323
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    • 2017
  • Geosmin and 2-methylisoborneol (2-MIB), responsible for earthy or musty smell, are a major concern for safe drinking water supplies. This study investigated the effects of environmental factors on odorous compound production and cell growth in cyanobacterial strains. Anabaena sp. FACHB-1384, a 2-MIB producer, was sensitive to low temperature (<$20^{\circ}C$). However, geosmin producers, Anabaena sp. Chusori and Anabaena sp. NIER, were sensitive to high light intensity (>$100{\mu}mol/m^2/sec$), but not to low temperature. Geosmin concentrations increased under higher nitrate concentrations, being linearly proportional to cell density. A P-limited chemostat showed that P-stress decreased the geosmin productivity and extracellular geosmin amount per cell in Anabaena sp. NIER. However, only 2-MIB productivity was reduced in Planktothrix sp. FACHB-1374 under P-limitation. The extracellular 2-MIB amount per cell remained constant at all dilution rates. In conclusion, high light intensity and P-stress can contribute to the lower incidence of geosmin, whereas 2-MIB reduction could be attainable at a lower temperature.

Improved Optimization of Indirubin Production from Bioreactor Culture of Polygonum tinctorium

  • Chung, Choong Sik;Kim, Kyung Il;Bae, Geun Won;Lee, Youn Hyung;Lee, Hyong Joo;Chae, Young Am;Chung, In Sik
    • Journal of Applied Biological Chemistry
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    • v.43 no.2
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    • pp.109-111
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    • 2000
  • Effect of the two-stage operation and cell concentration on indirubin production was investigated using bioreactor culture of Polygonum tinctorium. Two-stage culture was operated successfully for 110 days without any adverse effects on continuous indirubin production. Maximum indirubin concentration was found to be at 80 mg/bioreactor. Initial cell concentration significantly affected indirubin production. The indirubin production at 29.2% PCV was improved by 845%, compared to that at 5% PCV. For high-density bioreactor culture of P. tinctorium, a maximum production rate of 10.2 mg indirubin/L day was obtained. Indirubin recovery for bioreactor operation was also examined using XAD-2, XAD-4, XAD-7, and solid silicon. XAD-4 was 1.6-fold more effective than that for solid silicon in indirubin recovery.

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Isolation and In Vitro Culture of Vascular Endothelial Cells from Mice

  • Choi, Shinkyu;Kim, Ji Aee;Kim, Kwan Chang;Suh, Suk Hyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.1
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    • pp.35-42
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    • 2015
  • In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

Effects of Glucose concentration on the production of poly(3-hydroxybutyrate) by high cell density culture of Ralstonia eutropha

  • Shang, Longan;Kim, Do-Yun;Kim, Moon-Il;Kim, Byoung-Jin;Chang, Ho-Nam
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.213-217
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    • 2003
  • The effects of glucose concentration on the production of PHB by fed-batch culture of Ralstonia eutropha were investigated. In the range of glucose concentration of $2.5\;{\sim}\;40\;g/l$, it was found that the high glucose concentration was not favorable for the PHB formation after the phosphate limitation. It was further confirmed by the specific PHB synthesis rates and yields. The PHB concentration decreased much with the increase of glucose concentration. But if the glucose concentration was very low, e.g. 2.5 g/l, the cell growth and PHB synthesis also could be limited because of inadequate glucose supply. Itwould be better to maintain the glucose concentration at about 9.0 g/l to obtain high DCW, PHB concentration and productivity.

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Selection of Optimum Species of Tetraselmis for Mass Culture (대량배양에 적합한 Tetraselmis종의 선택)

  • 김철원;허성범
    • Journal of Aquaculture
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    • v.11 no.2
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    • pp.231-240
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    • 1998
  • Tetraselmis is widely used as a live food because of its easy handling, high nutrient, large size and wide tolerant range of temperature and salinity. In order to find the optimum Tetraselmis species for mass culture in Korea, five species of this microalgae were examined on size, optimum culture condition ${\textperthousand}$s, $^{\circ}C.$) and nutrient composition. The results obtained were as follows: Among five species of Tetraselmis, T. sp.(Haeundae) was the largest(major axis $17.6{\pm}1.87^{\mu}$m, mean cell volume 727${\mu}$m), and T. sp. (China) the smallest (major axis $14.6{\pm}1.46^{\mu}$m, mean cell volume 625m). Tetraselmis was very eurythermal and euryhaline species. But optimum temperature and salinity for growth were 24~$30^{\circ}C.$ and 27~30${\textperthousand}$, respectively. Among five species of Tetraselmis, T. sp. (China) seemed to be the most tolerant of high temperature over $30^{\circ}C.$, and T. tetrathele of low temperature below $6^{\circ}C.$. In culture density, T. suecica showed the highest growth rate among the among the five species. The cell density of this microalgae attained to $141{\times}10^4$cells/ml at $24^{\circ}C.$ and 30${\textperthousand}$ within 7 days. In chemical composition, crude protein amount was the highest in T. suecica (44.50%), and crude lipid amount it T. sp. (Haeundae, 7.13%). Total essential amino acid amount was the highest in T. sp. (Haeundae, 50.4%) and total polyunsaturated amount in T. sp. (China, 11.7%) The results on growth and chemical composition of five species of Tetraselmis indicated that T. suecica seemed to be the most suitable species for mass culture in Korea.

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Optimal Condition of Microporous Membrane for Bone Marrow Stromal Cell Allotransplantation to Stimulate Wound Healing in Vitro (창상치유목적의 골수기질세포 동종이식을 위한 고분자막의 조건)

  • Lee, Eun-Sang;Kim, Myeong-Joo;Han, Seung-Kyu;Hong, Sung-Taek;Kim, Woo-Kyung
    • Archives of Plastic Surgery
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    • v.37 no.5
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    • pp.509-518
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    • 2010
  • Purpose: Major drawbacks of conventional bone marrow stromal cells (BSCs) transplantation method are mainly caused by direct transplanted cell to host cell interactions. We hypothesized that separation of the transplanted cells by a microporous membrane might inhibit most of the potential adverse effects and induce superior effect. The purpose of the study is to determine the optimal condition of the microporous membrane. Methods: First, BSCs were placed in polyethylene terephthalate (PET) transwell inserts with 3, 8, or $12{\mu}m$ pore size, and cultured in 24 well culture plates. After 5 days, bottoms of the plates were observed for presence of attached BSCs in monolayer and cell numbers were evaluated. Second, BSCs were placed PET, polycarbonate (PCT), and mixed cellulose esters (MCE) transwell inserts with 3 and $8{\mu}m$ pore size, and cultured in 24 well culture plates. After 3 days, the supernatants of the media left in culture plate were analyzed for collagen, vascular endothelial growth factor (VEGF), platelet derived growth factor BB (PDGF-BB), and basic fibroblast growth factor (bFGF). Third, BSCs were placed in 15% and 70% of the PET membrane with $3{\mu}m$ pore size. All the experimental conditions and methods were same as the second study. Results: The optimal pore sizes to prevent BSC leakage were $3{\mu}m$ and $8{\mu}m$. The amounts of type I collagen and three growth factors tested did not show significant differences among PET, PCT, and MCE groups. However, the collagen, VEGF, and bFGF levels were much higher in the high (70%) density group than in the low (15%) density group. Conclusion: This study revealed that the optimal pore size of membrane to prevent direct BSC to recipient cell contact is in between $3{\mu}m$ and $8{\mu}m$. Membrane materials and pore sizes do not influence the collagen and growth factor passage through the membrane. The most striking factor for collagen and growth factor transport is pore density of the membrane.

Induction of the T7 Promoter Using Lactose for Production of Recombinant Plasminogen Kringle 1-3 in Escherichia coli

  • Lim, Hyung-Kwon;Lee, Shi-Uk;Chung, Soo-Il;Jung, Kyung-Hwan;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.225-230
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    • 2004
  • A plasminogen kringle domain 1 to 3, rKl-3, was expressed in Escherichia coli under the control of T7 promoter. For the cost-effective production of rKl-3, the induction process was analyzed and optimized. Induction characteristics with lactose were analyzed in terms of induction time and inducer concentration in various culture conditions including batch and high-cell-density fed-batch cultures. In the fed-batch culture, the induction around 6 h after initiation of the DO-stat fed-batch culture resulted in the highest expression level of rKI-3 among the induction points examined. The highest demand of oxygen at this point was crucial for the maximum expression level of rKI-3. As the lactose concentration increased, the expression level also increased, though the expression level showed a plateau above a concentration of 14 mM of lactose. Lactose acted less specifically than IPTG since most of it was hydrolyzed to glucose and galactose. However, using lactose, the cell growth and the maximum expression level of rKl-3 increased by 20% and 24%, respectively, compared with those using IPTG in the fed-batch culture. The lactose seemed to be hydrolyzed by intracellular and extracellular $\beta$-galactosidase liberated by cell lysis at the same time. Residual concentration of glucose was maintained to a a limit of detection by high performance liquid chromatography, and galactose was not consumed by the host strain Escherichia coli BL2l(DE3).

참당귀(Angelica gigas Nakai) 현탁세포 perfusion 배양 연구

  • Kim, Yeong-Hwa;Lee, Yong-Il;Kim, Ik-Hwan;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.301-304
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    • 2002
  • Perfusion culture strategies for high density culture of plant cell suspensions to enhance the productivity of extracellular polysaccharides were investigated. Angelica gigas Nakai cell suspensions were used to produce the extracellular polysaccharide and perfusion parameters were optimized to maximize the production. When the medium exchange was started at the fifth day after inoculation, the maximum cell concentration (23.8 g dry cell weight per liter) was achieved.

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Cultural Characteristics of Antagonistic Bacterium, Bacillus licheniformis N1 against Botrytis cinerea (잿빛공팜이에 대한 길항균 Bacillus Iicheniformis N1의 배양적 특성)

  • 이재필;문병주
    • Journal of Life Science
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    • v.11 no.2
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    • pp.173-180
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    • 2001
  • This study was conducted to estimate the cultural characteristics, the production of antibiotic, and the selection of optimal media for mass culture of Bacillus licheniformis N1 isolate which was previously reported as an antagonistic bacterium to Botrytis cinerea. We investigated initial pH, temperatures and shaking speed for good cultural conditions and antibiotics production by N1 isolate. According to the results, the optimal conditions of initial pH, temperatures, and shaking speed were determined to be pH 5.0~5.5, 30~35$^{\circ}C$ and 250 rpm, respectively. Also, the optimal conditions for the antagonism by N1 isolate highly appeared in the initial pH as 5.0, and the mycelial growth inhibition was high when the substances used such as glucose or corn starch as carbon sources, and biji(soybean curd residue) flour as a nitrogen source. Furthermore, inhibitory area was significantly expanded, when 3% or 5% of corn starch was added into 5% of Biji flour as nitrogen source, were respectivley selected for mass culture of N1 isolate. Among them, 5% Biji flour medium showed higher cell density more than 10 times that in NB medium after 48 hour incubation. Therefore, the optimal medium was determined as 5% biji flour added 3~5% of corn starch for high density of cells.

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