• Title/Summary/Keyword: Cell culture condition

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Characteristics of Growth and Oil Production of Peppermint Cells in an Air-bubble Bioreactor (기포 생물반응기에서 페퍼민트 세포의 생육 및 정유 생산 특성)

  • 송은범;이형주
    • KSBB Journal
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    • v.8 no.5
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    • pp.495-503
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    • 1993
  • To investigate the characteristics of growth and oil production of peppermint cells during a batch culture, cells derived from peppermint callus was cultivated in an air bubble reactor. During the batch culture, effects of inoculum size, abiotic stress, yeast elicitor, and two stage culture on the cell growth, the productivity of oleolesin, and the formation of flavor components were determined and also the sugar concentrations and kinetics of cell growth were analyzed. Among the various sizes of inoculum, the culture with 2.0% packed cell volume inoculum showed the optimum condition for cell growth in the proposed bioreactor, and the cell yield and essential oil production reached to 5.7g/1 and 0.109g/1, respectively. When the abiotic stress of daily 8hr dark and $10^{\circ}C$ cold treatments were given to the culture cell growth decreased but essential oil production increased to 0.546g/l. In a modified Lin-Staba medium in which 100mg/l yeast extract as an elicitor was added to the culture, the cell growth and oil production increased, and menthol content was 22.5% of oil. In the two stage culture, in which the basic culture conditions of 27$^{\circ}C$, light, and without elicitor were employed during the first six days followed by the second stage with daily 8hr treatment of cold and dark condition, and also with yeast extract as an elicitor, cell growth decreased after eight days, essential oil production was not increased, and menthol was not detected. Dry cell yield was 0.38g dry cell/g sugar and specific growth rate was 0.25 day-1. The major terpenoid in the oil was not the menthol but pulegone and piperitone, precursors of menthol were accumulated. However, when yeast elicitor was added, menthol was produced to the level of 22.5% which was the highest value in the peppermint cell culture reported so far.

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Isolation of Pseudomonas putida BM01 Accumulating High Amount of $PHA_{MCL}$

  • Song, Jae-Jun;Yoon, Sung-Chul
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.126-133
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    • 1994
  • A Pseudomonas putida strain able to accumulate high amount of polyesters of medium-chain-length 3-hydroxyalkanoic acids ($PHA_{MCL)$) was isolated from soil in a landfill site using an enrichment technique. Culture condition of the isolated strain for polyester production in a one-step culture was optimized in a mineral-salts medium against pH and concentrations of ammonium sulfate, carbon source(e.g., octanoate), and phosphate. The optimal values for maximal cell growth and PHA accumulation were: pH; 7$\sim$8, $(NH_4)_2SO_4$; 8 mM, octanoate; 40 mM. The optimum temperature was in the range of $20\sim30^{\circ}C$, which was rather broader than in other bacteria. Cell growth was strongly inhibited by the phosphate limitation to less than 1 mM. An increase of phosphate concentration above 1 mM showed little effect on cell growth and polyester accumulation. When the strain was grown on octanoate under this optimized condition it produced 3.4 g dry biomass per liter and yielded 1.7 g PHA per liter amounting to 53 wt% of dry cells. The monomer units composing the polyester synthesized from octanoate were 3-hydroxyoctanoate (3HO), 3-hydroxycaproate (3HC), and 3-hydroxybutyrate (3HB) (85:13:2, mole ratio). Other low linear $C_3\simC_{10}$ monocarboxylic acids were also tested for polyester production.

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Influence of light intensity and photo-bioreactor design for photo biological hydrogen production by Rhodobactor sphaeroides (Rhodobactor sphaeroides의 수소생산에 미치는 광세기 및 광합성 배양기 영향)

  • Kim, Mi-Sun;Baek, Jin-Sook
    • Transactions of the Korean hydrogen and new energy society
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    • v.15 no.2
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    • pp.166-174
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    • 2004
  • Purple non-sulfur bacteria, Rhodobacter sphaeroides KD131 grew to reach the maximum cell concentration in 45 hrs of incubation in the synthetic media containing (NH4)2SO4, L-aspartic acid and succinic acid as the carbon and nitrogen sources, respectively, at 30oC under 8 klux irradiance using halogen lamp. The strain produced hydrogen from the middle of the logarithmic growth phase and continued until the cell growth leveled out. The strain grew and produced hydrogen under the irradiance of 3-30 klux, but cell growth was inhibited over 100 klux. In addition, anaerobic/light culture condition was better than the aerobic/dark on the hydrogen production. Among various photo-bioreactors examined, the flat-vertical reactor manufactured using clear acrylic plastic material showed the best hydrogen production rate at the given culture condition.

Characterization of HEK293 and Namalwa Cell Cultures by Using Design of Experiment (실험계획법을 이용한 HEK293 및 Namalwa 세포배양 특성 규명)

  • Kang, Kyung-Ho;Seo, Joon-Serk;Kim, Dong-Il
    • KSBB Journal
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    • v.27 no.3
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    • pp.186-194
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    • 2012
  • Various human host cell lines, which are more effective than the other original human cell lines, have been developed and used. Highly efficient human cell line can be obtained from the fusion between human embryonic kidney 293 (HEK293) and human Burkitt's lymphoma cells (Namalwa). Fused cell line has the advantages of both cell lines such as the high transfection efficacy of HEK293 cells and the constitutive expression of Epstein-Barr virus (EBV) genome which is related with high expression of target protein and anti-apoptotic growth of Namalwa cells. In this study, characterization of two original cell lines was performed by using design of experiment (DOE) considering cell maintenance, media development, optimization of culture condition, and scale-up. The formation of aggregates was apparent with high glutamine concentration at more than 6 mM. Supplementation of hydrolysates showed positive effects on the growth performances of HEK293 cells. On the contrary, Namalwa cells showed negative results. It was confirmed that Namalwa cells were more sensitive to lower temperature at $35^{\circ}C$ and hyperosmotic condition over 260 mOsm/kg. In addition, both cell lines showed limited growth in 3-L bioreactor due to shear stress.

Production of Corydalis Alkaloids by Plant Cell Culture(I) (식물세포배양에 의한 Corydalis Alkaloid의 생산(I))

  • Chang, Jung-In;Shin, Seung-Won;Chi, Hyung-Joon
    • Korean Journal of Pharmacognosy
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    • v.26 no.4
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    • pp.419-425
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    • 1995
  • Corydalis remota Fish. ex Max. (Papaveraceae) is a well known medicinal plant being used as analgesics or anticonvulsive in oriental medicine. As the alkaloid content is known to vary depending on the environmental factors, the technology of plant tissue culture can be adopted as source of Corydalis-alkaloids. The present study describes an establishment of tissue cultures of Corydalis which produce alkaloids consistently. Callus were induced from immature seeds of Corydalis remota by placing the seeds on MS static media containing NAA(0.25, 1.0 and 4.0 mg/l, respectively). The combined treatment of NAA(1.0 mg/l) with cytokinin(BAP 0.5 mg/l) improved the induction of callus. TLC scanning data followed by sequential extraction and purification revealed that the induced callus contains a significant amount of alkaloids. Cell suspension cultures were established by transferring the induced callus into the liquid media with the same condition of plant growth regulators as the callus culture.

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Differentiation of CD31-Positive Vascular Endothelial Cells from Organoid Culture of Dental Pulp Stem Cells

  • Seo, Eun Jin;Park, Jae Kyung;Jeong, Hoim;Kang, Jung Sook;Kim, Hyung-Ryong;Jang, Il Ho
    • International Journal of Oral Biology
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    • v.43 no.2
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    • pp.77-82
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    • 2018
  • The mesenchymal stem cells (MSCs) that reside in dental tissues hold a great potential for future applications in regenerative dentistry. In this study, we used human dental pulp cells, isolated from the molars (DPCs), in order to establish the organoid culture. DPCs were established after growing pulp cells in an MSC expansion media (MSC-EM). DPCs were subjected to organoid growth media (OGM) in comparison with human dental pulp stem cells (DPSCs). Inside the extracellular matrix in the OGM, the DPCs and DPSCs readily formed vessel-like structures, which were not observed in the MSC-EM. Immunocytochemistry analysis and flow cytometry analysis showed the elevated expression of CD31 in the DPCs and DPSCs cultured in the OGM. These results suggest endothelial cell-prone differentiation of the DPCs and DPSCs in organoid culture condition.

Studies of Cyclosporin A Biosynthesis under the Conditions of Limited Dissolved Oxygen or Carbon Source in Fed-batch Culture (용존산소 제한 또는 탄소원 제한 조건의 유가식배양에서의 Cyclosporin A 생합성 연구)

  • 전계택;박성관;권호균;정연호;정용섭;장용근;이영행
    • KSBB Journal
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    • v.13 no.2
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    • pp.203-208
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    • 1998
  • We investigated the effects of dissolved oxygen (D.O.) and fructose (C-source) on cell growth and biosynthesis of cyclosporin A (CyA) produced as a secondary metabolite by a wild-type filamentous fungus, Tolypocladium inflatum. This was performed by controlling the level of D.O. and the residual C-source, as required, through adjustment of medium flow rate, medium concentration and agitation rate in fed-batch cultures. CyA production was furned out to be maximal, when D.O. level was controlled around 10% saturated D.O. and concentration of the C-source was maintained sufficiently low (below 2 g/L) not to cause carbon catabolite repression. Under this culture condition, we obtained the highest values of CyA concentration (507.14 mg/L), Qp (2.11 mg CyA/L/hr), $Y_x/s$ (0.49 g DCW/g fructose), $Y_p/s$<(22.56 mg CyA/g fructose), and YTEX>$_p/x$ (48.31 mg CyA/g DCW), but relatively lower values of cell concentration (11.98 g DCW/L) and cell productivity (0.043 g DCW/L/hr), in comparison with other parallel fed-batch fermentation conditions. These results implied that, in the carbon-limited culture with 10% saturated D.O. level, the producer microorganism utilized the C-source more efficiently for secondary metabolism.

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Mouse Granulocyte-marcrophage Colony-stimulating Factor Enhances Viability of Porcine Embryos in Defined Culture Conditions

  • S. H Jun;X. S Cui;Kim, N. H
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.71-71
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    • 2003
  • Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifunctional cytokine that has been implicated in the regulation of pre-implantation embryo development across several species. The aim of this study was to determine the effects of mouse granulocyte-macrophage colony-stimulating factor (mGM-CSF) on development of porcine parthenotes and nuclear transferred embryos, and on their expression of implantation-related genes. In the presence of bovine serum albumin, mGM-CSF did not increase the percentage of oocytes that developed to the blastocyst stage and at day 7 did not increase oocyte cell number. Addition of 10 mM GM-CSF to protein-free culture medium significantly increased the compaction and blastocoel formation of 1- to 2-cell parthenotes and cloned embryos developing in vitro. However, cell number was not increased when they were cultured in the presence of GM-CSF. Semi-quantitative reverse transcripts polymerase chain reaction (RT-PCR) revealed that mGM-CSF enhances mRNA expression of the leukemia inhibitory factor receptor, but does not influence interleukin-6 or sodium/glucose co-transporter protein gene expression in blastocyst stage parthenotes. These results suggest that mGM-CSF may enhance viability of porcine embryos developing in vitro in a defined culture medium.

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Zinc Restored the Decreased Vascular Smooth Muscle Cell Viability under Atherosclerotic Calcification Conditions

  • Shin, Mee-Young;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.19 no.4
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    • pp.363-366
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    • 2014
  • Zinc is considered to be involved in maintaining healthy vascular condition. Atherosclerotic calcification of vascular smooth muscle cells (VSMCs) occurs via the mechanism of cell death; therefore, cell viability is a critical factor for preventing VSMC calcification. In this study, we tested whether zinc affected VSMC viability under both normal physiological non-calcifying (0 mM P) and atherosclerotic calcifying conditions (3 and 5 mM P), since VSMC physiological characters change during the VSMC calcification process. The study results showed that an optimal zinc level ($15{\mu}M$) restored the decreased VSMC viability which was induced under low zinc levels (0 and $1{\mu}M$) and calcifying conditions (3 and 5 mM P) at 9 and 15 days culture. This zinc-protecting effect for VSMC viability is more prominent under atherosclerotic calcifying condition (3 and 5 mM P) than normal condition (0 mM P). Also, the increased VSMC viability was consistent with the decreased Ca and P accumulation in VSMC cell layers. The results suggested that zinc could be an effective biomineral for preventing VSMC calcification under atherosclerotic calcifying conditions.

Optimal Conditions for the Production of Immunostimulating Polysaccharides from the Suspension Culture of Angelica gigas Cells. (면역증강성 다당 생산을 위한 참당귀 세포배양의 최적조건)

  • 안경섭;서원택;심웅섭;김익환
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.130-136
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    • 1998
  • An Immunostimulating polysaccharide was produced from the suspension culture of Angelica gigas H4, plant cells. In order to enhance the polysaccharide production by the A. gigas cell culture, medium composition and physical conditions were optimized. Schenk and Hildebrandt (SH) medium was selected as an optimal basal medium for the growth of A. gigas. The maximum cell and polysaccharide concentration obtained in SH medium were 15.8 g DCW/l and 0.85 g polysaccharide/l, respectively, at $25^{\circ}C$ under dark condition. For the enhanced polysaccharide production, a polysaccharide production medium (PPM) was established by modifying Gamborg B5 medium with optimized carbon sources, growth regulators, organic and inorganic elements. Optimal initial pH and temperature were 6.0-6.6 and $20^{\circ}C$, respectively, and the dark condition was better than the light condition. The maximum polysaccharide concentration of 1.5 g/l could be obtained through the optimization of the medium composition and physical conditions.

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