• Proceedings of the Botanical Society of Korea Conference
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• 1995.06a
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• pp.79-90
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• 1995

Production of a cardiac glycoside, digoxin, by 12$\beta$-hydroxylation from digitoxin was studied in plant cell suspension cultures of Digitalis lanata. In order to increase the conversion yield, various culture conditions including immobilization were investigated and optimized. Since digoxin was released in the medium temporarily and converted further into a glucosylated product, deacetyllanatoside C, in situ adsorption of digoxin was employed to recover the product continuously. Amberlite resin XAD-8 showed the best adsorption characteristics for digoxin among the examined resins, and an integrated process was developed to increase the productivity. In addition, it was found that the utilization of $\beta$-cyclodextrin to entrap digoxin during the culture enhanced the biotransformation yield significantly.

• 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.

• KSBB Journal
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• v.18 no.6
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• pp.435-439
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• 2003

Plant cell culture can be divide into two classes non-organic culture and organic culture. Non-organic culture such as suspension culture has many researches, however organic culture about recombinant protein production has little researches. Recombinant protein produced through organ culture is quite stable and it can make proteins by itself without any grow regulators. Therefore organ culture is much easier than other methods. In this research, we used transformed tobacco seed. At first we germinated the seed then separated stems and leaves from the grown plant. And raised in liquid medium by in vitro vegetative reproduction. Continuing most suitable conditions, we compared the Quantities of recombinant protein from intra cellular with from extra cellular. And adding some permeabilizing agents (Pluronic F-68, Triton X-100, DMSO, PEG8000), we increased the productivity of the recombinant protein.

• Journal of Plant Biotechnology
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• v.4 no.2
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• pp.77-82
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• 2002

To elucidate the effect of sucrose on cell growth and anthocyanin production, 1, 3, 5, and 7% sucrose were applied to liquid MS basal medium supplemented with 0.5 mg/L BA + 0.1 and 1 mg/L 2,4-D. Higher sucrose concentration decreased the cell growth regardless of the hormonal composition. Cain in fresh weight was gradual, showing the peak at day 12 in culture, and then decreased. Anthocyanin content increased with sucrose concentration in the medium, and practically there was no difference in anthocyanin content between the two media differing in 2,4-D content. Sucrose concentration for appropriate anthocyanin production was 7%, while 5% was more suitable for increase in total anthocyanin content. At higher sucrose levels, anthocyanin content was high due to the cessation of the cell growth. Medium pH decreased at the early stage and gradually increased thereafter.

• KSBB Journal
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• v.14 no.5
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• pp.534-538
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• 1999

For the optimization of operating conditions for plant cell suspension culture in bioreactors, effects of bioreactor types, various kinds of impellers, and aeration rates were examined using Nicotiana tabacum cells as a model system. Stirred tank bioreactor and airlift bioreactor were used for the comparison of bioreactor type. Growth rates in both bioreactors were lower than in shake flasks. In terms of final cell concentration, stirred tank bioreactor supported a little bit better growth compared to airlift bioreactor. Impeller type did not affect cell growth significantly, but it was apparent that cell size index decreased in the case of using hollowed paddle impeller. When the aeration rate was maintained at 0.3 vvm, cell growth was the best. At above 1.0 vvm, growth inhibition as well a browning was noticed. In addition, it was found that cell size index reduced proportionally to the increased of aeration rate.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.287-292
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• 2005

Calli were induced by culturing the leaf segment of Actinidia deliciosa ${\times}$ A. arguta clone 118 on MS medium supplemented with 0.5 mg/L 2,4-D, 0.1 mg/L NAA and 0.05 mg/L BA for 8 weeks in light condition. The induced calli were inoculated in liquid MS medium containing 0.5 mg/L 2,4-D, 0.1 mg/L NAA, 0.05 mg/L BA and 3% sucrose to establish cell suspension culture. The cells at the exponential stage and the stationary stage could be observed between 5-11 days and after that 12 days in culture, respectively. The fresh weight of callus induced from the suspended cells did not vary much among the media containing eight different combinations of plant growth regulators tested. The highest frequency of shoot induction (88.3%) was observed in MS medium containing 2.0 mg/L zeatin. Either BA or zeatin mixed with thidiazuron (TDZ) seemed to be effective in shoot induction. The induced shoots were transferred to MS medium containing 0.2 mg/L zeatin for further shoot growth. And then the shoots were transferred to Standardi (ST) medium containing 1.0 mg/L indolebutyric acid (IBA) for rooting. Plantlets could be obtained through cell suspension culture of Actinidia deliciosa ${\times}$ A. arguta clone 118.

• Korean Journal of Plant Tissue Culture
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• v.27 no.4
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• pp.245-258
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• 2000

Somatic embryogenesis has become a major tool in the study of plant embryology, as it is possible in culture to manipulate cells of many plant species to produce somatic embryos in a process that is remarkably similar to zygotic embryogenesis. Traditionally, the process has been studied by an examination of the ex vitro factors which influence embryo formation. Later structural, physiological and biochemical approaches have been applied. Host recently, molecular tools are being used. Together, these various approaches are giving valuable information on the process. This article gives an overview of somatic embryogenesis by reviewing information on the morphogenesis, physiology, biochemistry and molecular biology of the process. Topics covered include a brief description of the factors involved in the production of embryogenic cells. Carrot cell suspension is most commonly used, and the development of a high frequency and synchronous system is outlined. At the physiological and biochemical lev-els various topics, including the reactivation of the cell cycle, changes in endogenous growth regulators, amino acid, polyamine, DNA, RNA and protein metabolism, and embryogenic factors in conditioned medium are all discussed. Lastly, recent information on genes and molecular markers of the embryogenic process are outlined. Somatic embryogenesis, the best example of totipotency in plant cells, is not only an important tool in studies in basic biology, but is potentially of equal significance in the micropropagation of economically important plants.

• Journal of Microbiology and Biotechnology
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• v.8 no.5
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• pp.478-483
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• 1998

For the enhanced production of a cardiac glycoside, digoxin, using in situ adsorption by biotransformation from digitoxin in plant cell suspension cultures, selection of proper resins was attempted and the culture conditions were optimized. Among various kinds of resins tested, Amberlite XAD-8 was found to be the best for digoxin production in considering adsorption characteristics as well as the effect on cell growth. Adequate time for resin addition was determined to be 36 h from the beginning of biotransformation and the presence of resins should be as short as possible to increase the productivity. In addition, to prevent the cells from direct contact with resin particles, immobilized systems were designed and examined. Immobilization further improved the advantages of in situ adsorption. It was confirmed that the increase of the contact area for mass transfer was an important factor in utilizing an immobilized system to enhance digoxin production.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.143-148
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• 1995

The competence of callus formation and plant regeneration from root derived callus was higher in japonica cultivars than those of Tongil-type cultivars of rice. A japonica type cultivars Yeongdeogbyeo, showed the highest capacity (13%) for plant regeneration from root calli of 6 cultivars tested. The callus induced from seed and root tissues maintained higher capacity for plant regeneration during 7 passages of subculture on N$_{6}$ solid media at 2-week intervals. The maximum frequency (2 x 10$^{5}$ mL) of round cells and their cell colonies showed about 24 days after suspension culture of root-derived callus in N$_{6}$ medium with lmg/L 2,4-D, 300mg/L casein hydrolysate, 10mM L-proline, 20g/L sucrose and 30g/L sorbitol. The frequency of somatic embryo formation in suspension cultures of root-derived callus increased with prolonged advance of subculture time from 30 to 90 days, but their regenerative capacities decreased.

• Journal of Microbiology and Biotechnology
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• v.17 no.12
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• pp.1944-1948
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• 2007

Silkworm hemolymph (SH), prepared from fifth-instar larvae of Bombyx mori and heat-treated at $60^{\circ}C$ for 30 min, was used to improve cell viability and the production of human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) in transgenic Oryza sativa L. cell suspension cultures. Even though SH could not elevate cell viability at the concentrations up to 3% (v/v), addition of 0.3% (v/v) SH to a culture medium enhanced the production of hCTLA4Ig by 36.8% over an SH-free medium. Moreover, the production period of hCTLA4Ig could be shortened in a 0.3% (v/v) SH-added medium compared with that in an SH-free culture. As a result, addition of 0.3% (v/v) SH improved the productivity of hCTLA4Ig significantly in transgenic rice cell cultures.