• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.115-120
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• 1995

A method for cryopreservation of suspension cultured embryogenic cells derived from immature zygotic embryos of rice (Korean cultivars, Donggin-byeo and Taebaeg-byeo) was developed. The highest cell regrowth after storage in liquid nitrogen was obtained when Donggin-byeo cells were cryoprotected with a mixture of 2 M DMSO and 0.4 M sucrose and Taebaeg-byeo cells with a mixture of 0.64 M DMSO and 0.4 M sucrose at frequencies of 88% and 90%, respectively, Pretreatment in a high osmotic medium was not necessary. Upon transfer to $N_{6}$ medium suplemented with lmg/L NAA and 5 mg/L kinetin, the regenerated calli gave rise to numerous somatic embryos which subsequently underwent development into plantlets. Among approximately 100 plantlets, 25% of them were albinos.s.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.59-64
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• 1995

Effects of salicylic acid (SA) on anthocyanin synthesis in cell suspension cultures of grapes (Vitis vinifera L.) were investigated. tow concentrations (0.1 to 1$\mu$M) of SA did not affect the cell growth and anthocyanin accumulation whereas high concentrations (5 to 10$\mu$M ) of SA inhibited cell growth with increasement of anthocyanin synthesis. Five micromoles of SA promoted anthocyanin accumulation 4 folds compared to control cells. When SA was treated on the different culture times (0 to 7day), the highest pigment accumulation was obtained at the cells of second day. A low productivity of anthocyanin under continuous dark incubation was also recovered by adding SA which mimicked light irradiation effect These results suggest that SA is one of essential agents in anthocyanin biosynthesis.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.3
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• pp.187-191
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• 2003

Proteolytic enzymes existing in plant cell cultured media are the major reason for the loss of secreted human granulocyte-macrophage colony-stimulating factor (hGM-CSF). The addition of pepstatin, aprotinin and PMSF relatively decreased the proteolytic degradation of hGM-CSF in a conditioned medium, but sufficient prevention against the proteolytic activity could not be obtained with chemical protease inhibitors. Gelatin, as a competitive substrate for protease, showed a stabilizing effect in a conditioned medium. Compared to the initial hGM-CSF concentration in a conditioned medium. with 10 g/L of gelatin, 68% of the hGM-CSF remained after 5 days. In a cell culture experiment, 5 g/L of gelatin significantly stimulated the hGM-CSF production and accumulation in culture media, with no growth inhibition. compared to the controls (4.72 $\mu\textrm{g}$/L), the extracellular hGM-CSF level could be increased to 39.78 $\mu\textrm{g}$/L with the addition of 5 g/L of gelatin.

• Applied Biological Chemistry
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• v.35 no.6
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• pp.443-448
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• 1992

To investigate the production of monoterpenoids by Mentha pipperita cells in suspension culture, effects of media formulation, plant growth hormones, initial pH of the media, and cold stress on the production of essential oil and menthol were analyzed. Among the media employed, Lin-Staba medium resulted in the best essential oil production. Addition of 100 mg/l of yeast extract to the Lin-Staba medium induced the cells to produce large amount of essential oil and high content of menthol (0.39 g/l and 19.6%, respectively). In the effect of plant growth hormone, auxine were more effective than cytokinins. At initial pH of 4.7, oil production was good but menthol content was low. However at pH 5.7 the trend was reversed. When the culture temperature was lowered from $27^{\circ}$ to $10^{\circ}$ during 6 hour-dark period, growth was not changed much but essential oil production and menthol content was increased and reached to 528 mg/l and 21%, respectively.

• Korean Journal of Pharmacognosy
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• v.16 no.3
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• pp.171-174
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• 1985

Panax ginseng root has been widely used as an important drug for thousands years in China, Korea and Japan. The main effective components of ginseng have been believed to be saponins. However, ginseng cultivation is very difficult and needs many years for growth. It has already been shown that Panax ginseng callus produces a considerable amount of the same kinds of saponins as in intact plants. Various culture conditions were examined for increased production of ginseng saponins by cell culture. The saponin contents and the growth rates in two cell lines of ginseng callus were compared in static and suspension cultures, rotary and reciprocal shaking cultures. It was shown that the growth rate in rotary shaking cultures of D5-B2K-B2K callus was the highest and ginseng saponin production was most effective in reciprocal cultures of D5-B2K-B2K callus. The saponin content per fresh weight of the culture was 1.03 times higher than that of the fresh ginseng root.

• 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.

• KSBB Journal
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• v.31 no.4
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• pp.284-290
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• 2016

In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.442-448
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• 2006

Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.1
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• pp.72-74
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• 2001

Transgenic Nicotiana tabacum cells were cultivated for the production of murine granulocyte macrophage-colony stimulating factor (mGM-CSF) in both a stirred tank bioreactor and an airlift bioreactor with draft tube. Cell growth and mGM-CSF production in the airlift bioreactor were found to be better than those achieved in the stirred tank bioreactor. In the airlift bioreactor, 9.0g/L of cells and 2.2ng/mL of mGM-CSF were obtained (11.0g/L and 2.4ng/mL, respectively in shake flasks). Although the lag period was prolonged and mGM-CSF production was lowered by 33% in the stirred thank bioreactor as compared to the control culture, the maximum cell density was increased up to 12.0g/L due to better mixing by agitation at the higher cell density.

• Natural Product Sciences
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• v.7 no.4
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• pp.120-123
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• 2001

To develop systems for economic production of useful essential oil compounds, callus was induced from the seedlings of Agastache rugosa and cultured on MS medium. The volatile oil fraction was extracted from the callus and investigated by mean of GC-MS. The composition of the oil was compared with that of the mother plant. As a result, sixty five compounds including ferruginol were identified in the essential oil fraction. The main component of the oil from the leaves of Agastache rugosa was methyl chavichol (53.6%). Methyl jasmonate and jasmonic acid were added to the culturing cell suspension, separately and the composition of induced oil were compared. The oils from cultured cells treated with jasmonates showed considerably different patterns. Especially, the peak of estragole was found in callus oil after treatment with methyl jasmonate as though the amount was limited to 0.58%. In general, the TIC pattern of GC-MS of the callus oil became more similar to the oil from the leaves after elicitation.