• Plant Biotechnology Reports
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• v.5 no.2
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• pp.127-134
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• 2011

The biotransformation potential of cell suspension cultures generated from Withania somnifera leaf was investigated, using withanolides, i.e. withanolide A, withaferin A, and withanone as precursor substrates. Interestingly, the cell suspension cultures showed inter-conversion of withanolides, as well converted to some unknown compounds, released to the culture media. The bio-catalyzed withanolide was detected and quantified by TLC and HPLC, respectively. There is noticeable conversion of withanolide A to withanone, and vice versa though at a lower level. The type of reaction of this biotransformation appears to be substitution of 20-OH group to 17-OH in withanolide A. In this paper, we present for the first time the possibility of biotransformation by inter-conversion of withanolides of pharmacological importance through cell suspension culture of W. somnifera. The possible role of putative cytochrome $P_{450}$ hydroxylases is implicated in the conversion.

• Natural Product Sciences
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• v.3 no.1
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• pp.71-74
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• 1997

Cell suspension cultures of Solanum mammosum transformed inoculated salicyl alcohol into salicin $(salicyl\;alcohol\;2-0-{\beta}-D-glucopyranoside)$. The highest level of salicin (59.3 mg/flask) in the cells was formed within 3 days after inoculating with salicyl alcohol (50 mg /flask containing 50 ml medium). The glucosylation capability of salicyl alcohol by cell suspension cultures of S. mammosum was relatively higher than that reported previously.

• Korean Journal of Plant Resources
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• v.24 no.3
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• pp.280-285
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• 2011

In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.1
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• pp.51-55
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• 2001

This study was conducted to establish a plant cell culture system for the production of medically important secondary metabolites from Xanthium strumarium. The effects of plant growth regulators including NAA, 2,4-D, kinetin, and ABA were examined in terms of callus induction, maintenance of callus and suspension cultures. It was shown that callus was induced upon treatment with NAA while embryo was induced after treatment with 2,4-D. Callus formation was further improved by treatment with ABA and NAA. The level of callusing increased by 17-29% for the seed case, cotyledon, leaf, and hypocotyl and by 96% in the case of the root. Suspension cell lines were established using calli produced from cotyledon, hypocotyl and root and cultured at 25$\^{C}$ under light conditions. The cells grew up to 15g/L with NAA 2ppm, BA 2ppm, and ABA 1ppm treatment. Supernatants of suspension cultures of cell lines derived from coyledon and hypocotyl produced some distinctive secondary metabolites, one of which was identified as 8-epi-tomentosin, which belongs to the xanthanolides. The amounts of 8-epi-tomentosin produced by the cotyledon- and hypocotylderived cell lines were 13.4mg/L and 11.0mg/L, respectively.

• KSBB Journal
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• v.13 no.1
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• pp.26-30
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• 1998

The suspension cultures of Mentha piperita produce menthol which has very low solubility in water due to its hydrophobicity. This can be considered as a factor for its low production in the suspension suspension cultures. Cyclodextrin has the hydrophobic cavity inside the molecule in which menthol can be captured and allow to form a stable complex. The suspension culture of Mentha piperita showed 70% higher production enhancement in the medium containing 1.5%(w/v) $\beta$-cyclodextrin than the control. $\beta$-cyclodextrin had no adverse effect on the cell growth and showed the best result among $\alpha$-, $\beta$- and $\gamma$-cyclodextrins tested in terms of menthol production. We demonstrated that $\beta$-cyclodextrin can be used to enhance the production of menthol in the suspension cultures by capturing hydrophobic menthol into the cavity of cyclodextrin molecules.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.154-159
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• 2006

Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.

• KSBB Journal
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• v.19 no.5
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• pp.374-380
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• 2004

Production of therapeutic proteins by transgenic plant cell suspension cultures is an attractive system alternative to the other expression system. However, plant cell cultures have shown low expression level of foreign proteins and decreased cell viability by the changes of culture conditions. Therefore, it is necessary to enhance cell viability during the culture period. In this study, a quantitative analysis technique was designed to measure relative cell viability for plant suspension cells which have cell wall and aggregates. It was found that the programmed cell death of plant cells by apoptosis was essentially linked with the apoptotic pathway of animal cells. Therefore, effects of nicotinamide, 3-aminobenzamide and antioxidants on cell viability and apoptosis were examined in transgenic Nicotiana tabacum cells producing hGM-CSF. With those additives, cell viability could be maintained and apoptosis could be redued. In the result, the extracellular production of hGM-CSF could be enhanced 2.5 fold. It was also found that the supplementation of glutathione and ascorbic acid suppressed both the cold stress-induced decrease in cell viability and the increase of total genomic DNA fragmentation.

• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.228-235
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• 2010

The establishment of cell suspension culture and plant regeneration of the halophytic Leymus chinensis (Trin.) are described in this study for the first time. Callus induction solid medium containing Murashige and Shoog (MS) basic salt, $2.0\;mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), and $5.0\;mg\;l^{-1}$ L-glutamic acid with $30.0\;g\;l^{-1}$ sucrose and $4.0\;g\;l^{-1}$ gelrite for solidification induced the highest rate of cell division in Type 1 callus among calli of various types. Liquid medium with the same hormone distribution was therefore, used for cell suspension culture from Type 1 callus. Over a 30 d suspension culture at 100 rpm, great amounts of biomass were accumulated, with 71.07% average daily increment and 22.32-fold total fresh weight increment. Comparison of before and after suspension culture, the distribution of different size callus pieces and the maintenance of callus type were basically unaltered, but a slight increase in relative water contents was observed. To induce the potential of plant regeneration, the directly transferring on plant regeneration solid medium containing MS basic salt, $0.2\;mg\;l^{-1}$ $\alpha$-naphthalene acetic acid (NAA), $2.0\;mg\;l^{-1}$ kinetin (Kn), and $2.0\;g\;l^{-1}$ casamino acid and indirectly transferring were simultaneously performed. Even now growth rates of suspension-derived callus on solid medium were approximately half of those of Type 1 callus, but faster somatic embryogenesis was observed. Rooting of all regenerated shoots was successfully performed on half-strength MS medium. All plants appeared phenotypically normal.

• Toxicological Research
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• v.11 no.1
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• pp.103-108
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• 1995

Rat renal proximal tubule suspension was prepared from adult male Sprague Dawley rat (250-300g) by mechanical (non-enzymatical) method and evaluated as a pontential model for mechanistic studies and early screening of nephrotoxicity, using anionic antibiotics (cephaloridine). Cephaloridine (CPL) produced an increase in LDH release into media. This release results from decrease a proximal tubule cell viability and subsequently increase the permeability of cell viability and subsequently increase the permeability of cell membrane. Since loss of intracellular potassium and ATP into media is the sign of disruption of cell membrane, especially basolateral membrane (BLM), CPL induced proximal tubule cell compromise also appear be associated with BLM, maybe $Na^+-K^+$ ATPase. Also seen was significant depression in brush border membrane (BBM) ALP activity and no significantly increase in BBM GGT activities. The inhibition of typical anion, PAH accumulation (especially, CPL 5 mM) and cation, TEA (especially, 4hours incubation) were seen dose dependently. This is because of CPL accumulation in renal proximal tubule and increase of cytotoxicity.

• Journal of Plant Biotechnology
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• v.6 no.4
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• pp.265-268
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• 2004

Cell growth and ginseng saponin production by large-scale suspension (bioreactor) cultures of Panax ginseng were investigated under various inoculum sizes. Cell growth was low at an inoculum size of 40 g FW/L, and the maximum cell growth was obtained with increasing inoculum size up to 100 g FW/L. The cell density of 333 g FW/L and 12.7 g DW/L was obtained at inoculum size of 100 g FW/L after 30 days of cultivation. Maximum saponin production of $4.40\;\cal{mg/g}$ DW was achieved at 60 g FW/L of inoculum size. Thus, inoculum size 60 g FW/L was suitable for optimum biomass accumulation as well as saponin production during bioreactor cultivation of ginseng suspension cells.