• Title/Summary/Keyword: suspension cultured cell

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Isolation and Culture of Protoplasts Derived from Embryogenic Cell Suspension Culture of Oryza sativa (Rice) (벼 진탕 배 배양세포로부터 원형질체 분리 및 배양)

  • Hwang, Baik;Kim, Mee-Kyung;Vasil, I. K.
    • Journal of Plant Biology
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    • v.31 no.1
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    • pp.41-49
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    • 1988
  • Several cultivars of rice were examined for induction of embryogenic callus on a medium containing MS salts, vitamins and 2, 4-D under darkness. Embryogenic callus was obtained from cultivar Cheonma with high ratio and embryo-like structures were formed from the callus on a medium with or without reduced 2, 4-D. Somatic embryoids with a plumule and radicle axis surrounded by a scutellum were observed. These embryoids germinated and produced plantlets in 30 days on the same medium. Protoplasts isolated from an embryogenic cell suspension culture derived from embryogenic callus were cultured either in liquid or in agar medium and protoplast derived cell colonies were obtained in 3-4 weeks.

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Production of Volatile Oil Components by Cell Culture of Agastache rugosa O. Kuntze

  • Shin, Seung-Won;Kim, You-Sun;Kang, Chan-Ah
    • 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.

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High frequency plant regeneration from zygotic-embryo-derived embryogenic cell suspension cultures of watershield (Brasenia schreberi)

  • Oh, Myung Jin;Na, Hye Ryun;Choi, Hong-Keun;Liu, Jang Ryol;Kim, Suk Weon
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.87-92
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    • 2008
  • An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when cultured on halfstrength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly with increasing concentrations of 2,4-D up to $3mg\;l^{-1}$, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryoderived white friable callus were established using half-strength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of water-shield, which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could be useful for the mass propagation and transformation of selected elite lines.

Plant Regeneration from Single Cell Culture of Wheat (Triticum aestivum L.) (밀(Triticum aestivum L.)의 단세포 배양에 의한 식물체의 재분화)

  • 김시철
    • Journal of Plant Biology
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    • v.32 no.4
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    • pp.227-233
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    • 1989
  • Single cells obtained from suspension culture of mature embryo-derived callus in wheat(Triticum aestivum L. cv Jang Kwang) were cultured to regenrated into the plantlet. Cell clusters and embryogenic calluses were efficiently developed from when the single cells clutured on the MS medium supplemented with 10${\mu}{\textrm}{m}$ 2,4-D. Upon transfer to hormone-free MS medium containing 10 mg/I AgNO3, embryogenic calluses gave rise to shoots, probably through somatic embryogenesis.

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Effects of Several Elicitors and Amino Acids on Production of Taxane Derivatives in Cultured Cells (수종의 Elicitor 및 Amino Acid가 배양세포내 Taxane 유도체의 생산에 미치는 영향)

  • Lim, Sook;Shin, Seung-Won
    • Korean Journal of Pharmacognosy
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    • v.29 no.4
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    • pp.360-364
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    • 1998
  • To develop new elicitors inducing the high productivity of taxane derivatives, plant growth inhibitors, namely, maleic acid hydrazide, N-phosphomethyl glycine and succinic acid 2.2-dimethyl hydrazide, coconut milk and yeast extract were administrated in the cell suspension culture system of Taxus cuspidata, and the production of baccatin III were analysed. The effects of amino acid related with the biosynthesis of baccatin III were also examined in these culture system. As the results, a remarkable enhancement of baccatin III production was observed in the cultivation with coconut water and with maleic acid hydrazide.

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Digoxin Production by Using Biotransformation in Digitalis lanata Cell Suspension Cultures (Digitalis lanata 현탁세포배양에서의 생물학적 변환을 이용한 Digoxin 생산)

  • 김혜경;홍희전;김동일
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.651-658
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    • 1994
  • For the production of digoxin by using biotransformation in suspension-cultured Digita- lis lanata cells, a two-stage culture process was optimized. Modified Murashige and Skoog medium was used for growth in the first stage and the cells were transferred to glucose solution for the production of digoxin from digitoxin via biotransformation in the second stage. When the cells were cultivated for 10 days in the growth period, 12$\beta$-hydroxylation capacity was the best. It was found that the optimum amount of digitoxin as substrate was 400 mg/l with initial cell density of 21%. Maximum productivity was achieved 5 days after transfer of cells to production medium. Sucrose and fructose provided similar digoxin yield as that in glucose, and 6% was proved to be the best glucose solution. Most of the components of modified MS medium except phosphate reduced the efficiency of digoxin formation. Besides, peptone and beef extracts inhibited 12$\beta$-hydroxylation, while promoting glucosylation. Finally, it was apparent that light enhanced the formation of digoxin significantly.

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Studies on the Production of Bioactive Substances -Callus Culture of Rehmanniae Radix-

  • Chi, Hyung-Joon;Kim, Hyun-Soo;Cho, Hi-Jae
    • Korean Journal of Pharmacognosy
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    • v.25 no.1
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    • pp.28-30
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    • 1994
  • The rate of growth and production of bioactive substances from Rehmannia glutinosa Liboschitz (Scrophulariaceae) were studied with the variation on the constituents of the culture media. The best growth was observed from MS basal medium containing 3.0 ppm NAA and 2.0 ppm kinetin. Carbohydrates (fructose, glucose and sucrose), phytosterols(${\beta}-sitosterol$, campesterol and stigmasterol) and carotenoid like substances were identified by GC-MS and TLC from the callus mass. However, catalpol was not detected from both solid and cell suspension cultures containing geraniol. Callus cultured Rehmannia glutinosa in the MS basal medium containing 0.1 ppm NAA and 0.1 ppm kinetin become differentiated to root.

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THE EFFECTS OF GINGIVAL FIBROBLAST ON THE MINERALIZATION OF THE RAT BONE MARROW STROMAL CELL (백서 골수세포의 석회화 과정에 미치는 치은 섬유아세포의 영향)

  • Kim, Seuk-Yong;Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.210-221
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    • 1995
  • The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization. For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco's Modified Eagle's Medium(DMEM, Gibco, U.S.A.). Every cell were cultured in state at $37^{\circ}C$, 100% of humidity, 5% of $CO_2$ incubator. Bone marrow stromal cells were isolated from 5-clay-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment. Cell were incubated into the 24 multi-well plate with $5{\times}10^4$cell/well of medium at $37^{\circ}C$, 100% of humidity 5% $CO_2$ incubator for 24 hours. After discarded of the supernatent of medium, O.5ml of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope. We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope. Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell. The results were as follows; The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only. The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal cell. It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.

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Effects of Dykellic Acid Derived from Microorganism on the Cell Growth and Superoxide Dismutase Activity in Tobacco Photomixotrophic Cultured Cells (미생물 유래 Dykellic Acid가 담배 녹색배양세포의 생장 및 Superoxide Dismutase 활성에 미치는 영향)

  • 곽상수;권혜경;권석윤;이행순;이호재;고영희
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.133-136
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    • 2000
  • To evaluate the biological effects of dykellic acid, a novel apoptosis inhibitor, isolated from microorganism on the plant cells, the cell growth, protein contents, and superoxide dismutase (SOD) activity were investigated in suspension cultures of tobacco photomixotrophic cultured (PM) cells on 12 days after different concentration of chemical treatment. The cells were cultured in MS medium containing 0.7 mg/L 2,4-D, 0.3 mg/L kinetin, 30 g/L sucrose and 200 mM NaCl at $25^{\circ}C$ in the light (100 rpm). Dykellic acid strongly inhibited the cell growth by evaluating the cell fresh wt and the ion conductivity in the medium ($IC_{50}$/, about 20 $\mu$M). The results as inhibition of cell growth and cell wall damage were same. The compound significantly increased the protein contents and the SOD specific activity in proportion with the dosage. The results suggested that dykellic acid may have biological activity in plant cells and tobacco PM cells may be suitable biomaterials for in vitro evaluation of the biological activity of natural products.

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