• Title/Summary/Keyword: Cell suspension cultures

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Optimization of Treatment Concentration and Screening of Exogenous Plant Growth Regulators for Improvement of Salidroside Yield in Rhodiola sachalinensis A. Bor Cell Suspension Cultures (참돌꽃 (Rhodiola sachalinensis A. Bor) 현탁세포배양에서 Salidroside 생산을 위한 외재 호르몬의 영향 및 처리 농도의 최적화)

  • Choi Hye-Jin;Kim Su-Jeong;Hwang Baik;Ahn Jun-Cheul
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.105-109
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    • 2005
  • To enhance salidroside productivity from a cell suspension cultures of Rhodiola sachalinensis, various combinations of auxin (NAA) and cytokinins (BA, Kinetin) concentration and addition of $GA_3$, TDZ, zeatin, spermine and spermidine at a hormone combination to be established were examined. NAA/BA combination is superior to NAA/kinetin combination in biomass and salidroside content. Maximum salidroside production ($64.6{\pm}8.9\;mg/L$ medium) was obtained from $2B_5$ medium with 1 mg/L NAA and 5 mg/L BA. The adding of $GA_3$ ($0.01\;mg/L{\sim}5\;mg/L$) was beneficial for salidroside accumulation and the highest productivity of salidroside, $90.3{\pm}8.34\;mg/L$, was obtained from $2B_5$ medium supplemented with 1 mg/L NAA, 5 mg/L BA and 0.1 mg/L $GA_3$. On TDZ, zeatin, spermine and spermidine, expectant results were not obtained except a little affirmative effect of spermidine ($69{\pm}2.88$ at 1 mg/L).

Systems for Production of Calves from Cultured Bovine Embryonic Cells (우 수정란의 배양세포들로부터 송아지 생산을 위한 체계)

  • ;N. L. First
    • Korean Journal of Animal Reproduction
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    • v.18 no.4
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    • pp.299-307
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    • 1995
  • The goal of cell stem cell technology is to produce a viable and genetically normal animal. To achieve this goal various laboratories have followed 2 different pathways beginning with either the culture of 1) single or pooled ICMs grown with or without a feeder layer or 2) single or pooled 16-20 cell stage embryos grown with a feeder layer. Also, thus far embryonic cell cultures or lines have been established by several methods including loose suspension culture for short-term cultures and more commonly murine or bovine fibroblast feeder layers for long-term culture. Pluripotent lines have been derived from 16-cell through blastocyst inner cell mass stages. The efficiency of establishing cell lines and cell proliferation apper to be affected by the number of cells or embryos starting the line. Most attempts to produce offspring from long term STO cell feeder layer cultured ICM or morulae derived ES cells have resulted in pregnancy failure in the first trimester when ES cells were used in cuclear transfer or have failed to retain ES cells in the progeny produced by chimerization. The exception is 1 chimeric fetus from use of morula ES cells in the chimerization with early embryonic cells. There is much to be learned yet about ES cell culture requirements for maintenance of totipotency. If bovine ES cell lines loose imprinting pattern and totipotency with long-term culture and passage as suggested for mouse ES cells, we may be limited to the use of short-term cultures for multiplication of embryos and efficient production of transgenic animals. No bovine ES cell system has yet met all of the criteria indicated for a totipotent ES cell line.

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Effects of Salt Concentrations on Accumulation of Pigments in Cell Suspension Cultures of Vitis vinifera and Phytolacca americana L. (포도와 미국자리공의 세포현탁배양계에 있어서 배지내 무기염 농도가 색소축적에 미치는 영향)

  • In, Jun Gyo;Lee, Young Bok;Choi, Kwan Sam
    • Korean Journal of Agricultural Science
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    • v.20 no.1
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    • pp.34-42
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    • 1993
  • Effects of salt concentrations on the cell growth and the pigment accumulation were investigated in cell suspension culture of Vitis vinifera and Phytolacca americana L.. The growth pattern of vine cell in control was showed the normal exponential growth pattern, but in the dilution media delay the exponential growth pattern from 4 to 8 days after culture. Maximal accumulation of anthocyanin was observed at 12 days after culture in all treatments. In cell suspension culture of Phytolacca, accumulation of betacyanin occurred in parallel with the cell growth pattern and maximal accumulation of betacyanin was observed after 8 days of culture. In the vine cell culture, the cell growth was showed the peak at 87.6mM of sucrose in the medium and reduced at over this concentration. Maximal anthocyanin accumulation was showed at 146mM of sucrose. In the higher concentrations of sucrose, the cell growth was rapidly decreased, but the accumulation of anthocyanin was not. Otherwise, in case of Phytolacca cell culture, betacyanin accumulation was showed in parallel with the cell growth increased with sucrose concentration. It was suggested that the anthocyanin of vine and the betacyanin of Phytolacca were controlled by different mechanisms.

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Somatic Embryogenesis and Plant Regeneration from Embryogenic cell Suspension Cultures of Schisandra chinensis Baill

  • Li, Cheng Hao;Niu, YudA;Zhao, Bo;Ghimire, Bimal Kumar;Kil, Hyun-Young;Heo, Kwon;Kim, Myong-Jo;Eom, Seok-Hyun;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.346-351
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    • 2007
  • An efficient somatic embryogenesis and plant regeneration protocol was developed for Schisandra chinensis Baill, using embryogenic cell suspensions and optimized media conditions. Friable embryogenic callus was induced from cotyledonary leaf and hypocotyl explants of 7 days old seedlings on MS agar medium supplemented with 1.0 to $4.0\;mg\;l^{-1}$ of 2,4-dichlorophenoxyacetic acid (2,4-D). Fast growing and well dispersed embryogenic cell suspensions were developed within two months when embryogenic calli were transferred to MS liquid medium containing $1.0\;mg\;l^{-1}\;2,4-D$. One third strength of MS medium was the best for both overall growth and development of somatic embryos in liquid culture. Over 3400 viable somatic embryos were produced from each 150 ml flask with an initial cell density of 30 mg in 30 ml medium. Germinated somatic embryos developed in liquid medium converted into plantlets after transferred to half-strength MS semi-solid medium. Approximately 90% of the converted plantlets were successfully transplanted to soil and grew into fertile plants.

Aloe 세포 배양계에서 $CO_2,\;O_2,\;C_2H_4$ gas가 세포 생장 및 다당체 생산에 미치는 영향

  • Park, Tae-Yeong;Jo, Gyu-Heon;Heo, Won;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.325-327
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    • 2000
  • We developed a on-line gas control system for three gas components and investigated the effect of oxygen, carbon dioxide, and ethylene on polysaccharide production and cell growth in the suspension cultures of Aloe saponaria. Carbon dioxide(5% v/v) enhanced significantly polysaccharide production while the cell growth was not affected.

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Increased Production of Digitoxin from Digitoxin by Biotransformation Using Plant Cell Culture

  • Hong, Hee-Jeon;Lee, Jong-Eun;Kim, Dong-Il
    • 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.

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형질전환된 Nicotiana tabacum 배양에 있어서 glutathione과 ascorbic acid가 세포생장과 생존율에 미치는 영향

  • Kim, Yong-Hun;Lee, Sang-Yun;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.293-297
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    • 2003
  • Glutathione and ascorbic acid have been shown to fulfill many essential functions in animal and plant growth, development, defence and protection against oxidative damage. Effects of glutathione and ascorbic acid were examined in transgenic N. tabacum cells producing hGM-CSF to determine the effects of the vitamins on growth and cell viability. In lag phase, cell viability was preserved by glutathione and ascorbic acid. Therefore, recombinant protein productivity was increased. The purpose of present study is to investigate the role of antioxidants in cold stress-induced apoptosis in plant suspension cells. Cold stress lowered cell viability and increased total genomic DNA fragmentation. Supplementing the cell cultures with glutathione and ascorbic acid inhibited cold stress-induced decrease in cell viability and increase in total genomic DNA fragmentation.

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Identification of Water Soluble Metabolites of Pentachlorophenol(PCP) in the Suspension Cultures of Soybean and Rice Cells;2. Isolation and characterization of PCP glucose conjugates (콩과 벼 현탁배양시(懸濁培養時) PCP 수용성대사물(水溶性代謝物)의 동정(同定);2. PCP glucose conjugates의 분리(分離) 및 분석(分析))

  • Kim, Pil-Je;Park, Chang-Kyu
    • Korean Journal of Environmental Agriculture
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    • v.15 no.1
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    • pp.37-45
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    • 1996
  • Abstracts From the previous metabolic study of Pentachlorophenol(PCP), PCP was found to be exclusively transformed into ${\beta}-glucose$ conjugates of PCP in soybean and rice cell suspension cultures. In order to gather structural information of of the glucose conjugate, their aglycons and glycon have been analyzed by GC and GC/MS respectively, after thorough purification by chromatographic techniques. The glucose conjugates were effectively purified through a 1-butanol extraction followed by Silica gel TLC, Sephadex column chromatography and HPLC. Aglycons of the metabolites were identified as PCP, isomeric mixture of tetrachlorophenol, and tetrachlorocatechol and glycon were identified as glucose, suggesting that there are at least three kinds of glucose conjugates with different phenolic moieties. Under controlled conditions, the glucose conjugates were separated into three HPLC peaks which released respective aglycon upon a hydrolytic treatment. These results give valuable information on the structure of the glucose conjugates such that some PCP-driven chlorophenols, in addition to PCP, are also conjugated with glucose.

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[ $Ce^{4+}$ ]-Stimulated Ion Fluxes Are Responsible for Apoptosis and Taxol Biosynthesis in Suspension Cultures of Taxus Cells

  • Li Jing-Chuan;Ge Zhi-Qiang;Yuan Ying-Jin
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.109-114
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    • 2005
  • Ion fluxes across the plasma membrane activated by 1 mM $Ce^{4+}$, cell apoptosis and taxol biosynthesis in suspension cultures of Taxus cusp/data were studied. The extracellular pH sharply decreased upon the addition of 1 mM $Ce^{4+}$, then increased gradually and exceeded the initial pH value over a time period of 12 h. The extracellular $Ca^{2+}$ concentration decreased within the first 3 h after the addition of $Ce^{4+}$, then gradually decreased to one third of initial value in control at about 72 h and remained unchanged afterwards. Experiments with an ion channel blocker and a $Ca^{2+}$-channel blocker indicated that the dynamic changes in extracellular pH and the $Ca^{2+}$ concentration resulted from the $Ce^{4+}$-induced activation of W uptake and $Ca^{2+}$ influx across the plasma membrane via ion channels. A pretreatment of the ion channel blocker initiated $Ce^{4+}$-treated cells to undergo necrosis, and the prior addition of the $Ca^{2+}$-channel blocker inhibited $Ce^{4+}$-induced taxol biosynthesis and apoptosis. It is thus inferred that W uptake is necessary for cells to survive a $Ce^{4+}$-caused acidic environment and is one of the mechanisms of $Ce^{4+}$-induced apoptosis. Furthermore, the $Ca^{2+}$ influx across the plasma membrane mediated both the $Ce^{4+}$-induced apoptosis and taxol biosynthesis.

Somatic Embryogenesis: Morphogenesis, Physiology, Biochemistry and Molecular Biology

  • Thorpe, Trevor A.
    • 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.

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