• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.325-328
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• 2002

Production of human granulocyte-macrophage colony stimulating factor (hGM-CSF) by Nicotiana tabacum cell suspension culture was studied in Murashige and Skoog (MS) medium with sucrose as a carbon source, ammonium nitrate and potassium nitrate as nitrogen sources, potassium dihydrogen phosphate and sodium dihydrogen phosphate hydrate as phosphate sources, respectively. Optimum concentrations for carbon, nitrogen, phosphate was determined to enhance the production of hGM-CSF. Cell growth was better at high initial sucrose concentration (60 g/L), high initial nitrogen concentration (121.04 mM). Maximum cell density (18.28 g/L) was obtained at 60 g/L of sucrose after 14 days. Cell growth was not so good at low initial sucrose concentration 00 g/L), but the highest hGM-CSF production was obtained at the latter half of exponential phase. hGM-CSF production increased about 3 fold at initial phosphate concentration of 4.96 nM

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.301-306
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• 2005

Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.183-188
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• 1997

Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

• Korean Journal of Plant Resources
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• v.24 no.4
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• pp.461-471
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• 2011

This study was carried out to establish an in vitro culture method of callus having a high antioxidant activity from Ginkgo biloba L. Leaf explants were cultured on Murashige and Skoog's medium supplemented with various growth regulators. The explants were incubated in the dark or 3,000 lux cool-white light. Methanol extracts from incubated callus were evaluated for scavenging activity of the free radicals using DPPH. The best callus growth rate was achieved in MS medium combined with 10 ${\mu}M$ NAA and 5 ${\mu}M$ kinetin in the light condition. Total antioxidant activity of cell aggregates in suspension culture [MS medium supplemented with 10 ${\mu}M$ NAA in the light] was up to 80% of ascorbic acid. By means of HPLC analysis, quantification of the quercetin dehydrate and keamperol profiles from suspension callus was compared. Contents of quercetin dehydrate and keamperol from leaf extracts were 0.07 and 2.24 ${\mu}g/20{\mu}l$, and those from callus 0.56 and 0.18 ${\mu}g/20{\mu}l$, respectively.

• Journal of Pharmacopuncture
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• v.13 no.3
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• pp.63-71
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• 2010

Objectives: Gallic acid (GA) is the major component of tannin which could be easily founded in various natural materials such as green tea, red tea, grape juice, and Corni Fructus. The purpose of this study is to investigate the effect of Gallic acid (GA) on production of interleukin (IL) in mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods: Productions of interleukins were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. Firstly, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, it was incubated with the antibody-conjugated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System and concentration of interleukin was determined. Results: The results of the experiment are as follows. 1. GA significantly inhibited the production of IL-3, IL-10, IL-12p40, and IL-17 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). 2. GA significantly inhibited the production of IL-6 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). 3. GA diminished the production of some cytokine such as IL-4, IL-5, and IL-13 in LPS-induced mouse macrophage RAW 264.7 cells. 4. GA did not show the inhibitory effect on the production of IL-$1{\alpha}$ and IL-9 in LPS-induced mouse macrophage RAW 264.7 cells. Conclusions: These results suggest that GA has anti-inflammatory activity related with its inhibitory effects on the production of interleukins such as IL-3, IL-10, IL-12p40, IL-17, and IL-6 in LPS-induced macrophages.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.4
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• pp.586-591
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• 2010

Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.

• Korean Journal of Acupuncture
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• v.27 no.3
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• pp.109-118
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• 2010

Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on chemokine and growth factor production in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Methods : Productions of chemokine and growth factor were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on xMAP$^{(R)}$ technology. Firstly, cell culture supernatant was obtained after treatment with LPS (1 ${\mu}g$/mL) and SB for 24 hours. Then, it was incubated with the antibody-conjugated beads for 30 minutes. Detection antibody was then added and incubated for 30 minutes. After incubated for 30 minutes, strepavidin-conjugated phycoerythrin (SAPE) was added. After another 30 minutes incubation, the level of SAPE fluorescence was analyzed in Bio-plex Suspension Array System. Results : The results of the experiment are as follows. 1. SB significantly inhibited the LPS-induced production of vascular endothelial growth factor (VEGF), interferon-inducible protein (IP)-10, and granulocyte-colony stimulating factor (G-CSF) at the concentration of 25, 50, 100, and 200 ${\mu}g$/mL in RAW 264.7 cells (P < 0.05). 2. SB significantly inhibited the LPS-induced production of Eotaxin at the concentration of 25, 100, and 200 ${\mu}g$/mL in RAW 264.7 cells (P < 0.05). 3. SB significantly inhibited the LPS-induced production of MIP-$1\alpha$ at the concentration of 25 and 100 ${\mu}g$/mL in RAW 264.7 cells (P < 0.05). Conclusions : These results suggest that SB has immuno-modulatory property related with its inhibition of VEGF, IP-10, G-CSF, and Eotaxin production in macrophages.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.57-61
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• 2000

We investigated the levels of glutathione (GSH) and its oxidized form (GSSG) in 24 cell lines derived from various plant species to understand the antioxidative mechanism in plant cell cultures. The total glutathione content was 98$\pm$27 $\mu$g/g cell fresh wt, showing a slight difference in plant species. The average contort of GSH and GSSG was 72$\pm$20 and 26$\pm$10 $\mu$g/g cell fresh wt, respectively. The average GSH content in plant cell lines occupies approximately 73% in total glutathione. During the suspension cultures of Scutellaria baicalensis, one of the plant species we tested, the GSH content decreased in proportion to the cell growth during the exponential growth stage, showing the low level at the stationary growth stage (84 $\mu$g/g cell fresh wt), whereas the GSSG content increased to the stationary growth stage (31 $\mu$g/g cell fresh wt). The results suggested that the ratio of GSH and GSSG should be involved in the cell growth and antioxidative mechanism in cultured cells.

• Journal of the Korean Society of Tobacco Science
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• v.1 no.1
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• pp.1-8
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• 1979

For the preliminary experiments of mass Production of tobacco cells in tank culture, the effects of nutritional conditions on the growth of suspended cells were investigated ; 1. The tobacco cell growth was affected by concentrations of sucrose or inorganic phosphate, type of nitrogen source, and plant hormone, especially 2, 4-D. 2. The optimum level of sucrose concentration was 3% and the level of inorganic phosphate was 0.3mg /ml, which was about twice as high as the level of Linsmaier - Skoog medium. 3. The best growth was observed when the ratio of nitrate nitrogen to ammonium nitrogen was 2 : 1, where the total nitrogen content was equal to that of nitrogen source. 4. To find out the mechanism of promotive effects of 214-D and inorganic phosphate on the tobacco cell growth, the respiration and metabolism of $^{14}\textrm{C}$-91ucose were investigated. Addition of 2, 4 -D in culture medium increased if 2, 4-D (0.2ppm )was added to medium or the level of inorganic Phosphate was raised 2.5 times as high as standard. In cultures with high inorganic phosphate and 2, 4-D, the absorbed 14C-glucose was converted to amino acids and organic acids rather than remained as sugars.

• Applied Biological Chemistry
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• v.38 no.5
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• pp.435-439
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• 1995

Some characteristics and pathogenicity of Hyphantria cunea nuclear polyhedrosis virus (HcNPV), a potential microbial pesticide was studied. H. cunea NPV replicated in the nucleus of S. frugiperda cells cultured in the TNMFH medium. In case of virus infected cell, prepolyhedra formation was observed at 24hrs post-infection. At 48 hrs post-infection, Most of the infected cell contained many mature polyhedra which were released into culture media 72 hrs post-infection, with the cells grown in suspension culture, pH of the culture medium increased during the virus replication: the pH of fresh medium was 6.35 and rose to 6.77 within 120 hrs. Polyhedra formed a band in linear density gradient of sucrose by centrifugation, which co-sedimented with $50{\sim}55%$ sucrose. The shape of the purified polyhedra was mostly tetragonal hexahedron and its size was about $2.5{\mu}m$. Electron microscopy and phase contrast microscopy showed that many bundled nucleocapsids were occluded in mature polyhedra at 48 hrs post infection. H. cunea larvae infected with NPV showed a higher motality in the second and third instar than in the fourth instar. Death rate of H. cunea larvae in the second and third instar fed with leaves coated with $1.5{\times}10^{9}{\sim}l.5{\times}10^{7}PIBs/ml$ reached more than 90%.