• Title/Summary/Keyword: Cell suspension cultures

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Effect of Osmotic Pressure on hCTLA-lg Production in Transgenic Rice Cell Suspension Cultures (형질전환된 벼세포 배양에 있어서 삼투압 조절에 따른 hCTLA4-lg 생산성 변화)

  • Choi Sung-Hun;Lee Song-Jae;Hong Seok-Mi;Cho Ji-Suk;Kim Dong-Il
    • KSBB Journal
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    • v.20 no.4
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    • pp.278-284
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    • 2005
  • An immunosuppressive agent, human cytotoxic T lymphocyte antigen 4 (hCTLA4), is used for the prevention of graft rejection and treatment of autoimmune diseases. hCTLA4-Ig, a CTLA4-immunoglobulin fusion protein, was produced and secreted from transgenic rice cell suspension cultures using rice a-amylase (RAmy3D) expression system. In this system, hCTLA4-Ig expression was regulated metabolically by sugar starvation. For the purpose of improving hCTLA4-Ig production, the effects of osmotic pressure was investigated in suspension cultures of transgenic rice cells. The highest production level was achieved at 40 mM sorbitol $(140\;mOsm{\cdot}kg^{-1}\;H_2O)$. Using the medium with 8 mM glucose, the level of hCTLA4-Ig in the medium reached 45.3 mg/L. By adjusting the osmotic pressure of induction medium, it was found that the hCTLA4-Ig production could be increased up to 2.1-fold compared with that in batch culture.

The Induction Time of Sucrose Active Transport System during the Phloem Cell Development in Suspension Cultures of Streptantus tortus Cotyledon (Streptanthus tortus 자엽의 배양세포에서 사부세포 발달동안 Sucrose 능동수송계의 유도 시기)

  • Cho, Bong-Heuy
    • Journal of Plant Biotechnology
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    • v.31 no.2
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    • pp.169-173
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    • 2004
  • Parenchyma cells of Streptanthus tortus suspension cultures possessed the different transport system for aldose-formed D-glucose and for ketose-formed D-fructose. $K_{m}$ value for D-glucose and D-fructose were 0.28mM and 15.02mM, respectively. $K_{m}$ value of D-mannose was 0.44 mM which is similar to the D-glucose transport system, but D-mannose was transported also through its own special uptake system. Parenchyma cells possessed the transport system of L-glucose, but the function of L-glucose was not known at all. Protoplast of parenchyma cells possessed only the monosugars transport system, but didn't possess the disugars, sucrose transport system. Early developing phloem protoplasts possessed glucose and sucrose transport system at the same time. On the contrary, in the complete developed phloem cells disappeared preexisted glucose transport system in the parenchyma cells, only new induced sucrose transport system existed.ted.

Comparative Characterization of Growth and Recombinant Protein Production among Three Insect Cell Lines with Four Kinds of Serum Free media

  • Kwon, Mi-Sun;Takashi Dojima;Park, Enoch Y.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.142-146
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    • 2003
  • Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

Plant Regeneration from Cryopreserved Embryogenic Cell Suspension Cultures of Korean Rice (Oryza sativa L.) Cultivars (한국 벼 품종 배발생 현탁배양 세포의 초저온 보존과 식물체 재분화)

  • 김석원;정원중;민성란;배경숙;유장렬
    • 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.

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Production of Artificial Seeds by Alginate-encapsulation of Rice Somatic Embryos (벼의 수화겔 인공종자 생산)

  • 정원중;민성란;송남희;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.183-186
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    • 1994
  • Somatic embryos derived from cell suspension cultures of rice were singly alginate-encapsulated to be used as artificial seeds. When placed on half strength MS solid medium,73% of the encapsulated somatic embryos were capable of germination Encapsulation per se did not affect the germination frequency of embryos. When incubated by wrapping with moistured non-sterile filter paper, 60% of the encapsulated somatic embryos germinated. However encapsulated zygotic embryos without endosperm showed a high germination frequency regardless of the sterility of the incubation conditions. The results suggest that a greater susceptibility of somatic embryos to contaminants is attributed to lower germination frequency of encapsulated somatic embryos in non-sterile conditions.

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Characterization of Cell Cultures and Ginsenoside Production by Cultured Ginseng and Wild Mountain Ginseng (산삼과 재배인삼의 세포배양 및 Ginsenoside 생성 특성)

  • 유병삼;장문식;변상요
    • KSBB Journal
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    • v.18 no.2
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    • pp.133-139
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    • 2003
  • Established cell-line cultures of cultured and wild mountain ginseng were characterized and their abilities to produce ginsenoside were determined. Cell lines were made of calli induced from the roots of wild mountain ginseng and cultured ginseng(Panax ginseng). Suspension cultures of wild mountain ginseng and cultured ginseng showed different growth and ginsenoside production rate. Their specific growth rates were 0.067 and 0.0035 day-1 in spite of having the same sugar consumption rates, where cells from wild mountain ginseng grew almost twice as fast as those of cultured ginseng. Their respective abilities to produce ginsenoside, however, were 0.53 and 2.53 mg/L.day, which means cells from cultured ginseng produced around 5 times more than wild mountain ginseng.

Production of 10-deacetylbaccatin III in Taxus cuspidata Suspension Cell Culture (주목 현탁세포배양을 이용한 10-deacetylbaccatin III 생산)

  • Lee, Gue-Wha;Kim, Dong-Il
    • KSBB Journal
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    • v.14 no.6
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    • pp.665-671
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    • 1999
  • In this study, enhanced production of 10-deacetylbaccatin III(10-DAB), a precursor of taxol in semisynthesis, was investigated in Taxus cuspidata cell suspension cultures. The effects of initial inoculum size and sugar concentration were examined to prove the relationship between the production of 10-DAB and cell growth. The cell growth was found to be stimulated in Schenk and Hildebrandt(SH) medium. The lower the inoculum size as well as initial sugar concentration, the faster the cell growth rate. When the initial sugar concentration was dept low, the production of 10-DAB into medium was increased. By using perfusion culture, continuous cell growth was possible until the end of culture and more than 34.67 g/L of cell concentration could by obtained. This is about 2.5 times higher level than that of control batch culture.

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Effects od Segree of Cell-Cell Contact on Liver Specific Function of Rat Primary Hepatocytes

  • Tang, Sung-Mun;Lee, Doo-Hoon;Park, Jung-Keug
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.99-105
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    • 2000
  • Cell-Cell interaction and the extracellular matrix (ECM) are belisved to play essential roles during in vitro culturing of primary hepatocytes in the control of differentiation and in the maintenance of tissue spcific functions. The objective of this study was to examine the effects of degree of cell-cell contact (DCC) on liver sperific function of rat promary hepatocytes. Hepatocyte aggregates with various with various degrees of cell-cell contantact, I. e., dispersed cell, longish aggregate, rugged aggregate, and smooth spheroid were obtained at 1, 5-6, 15-20, and 36-48 hrs, respectively in suspension cultures grown in spinner flasks embedded in Caalginate bead and collagen gel in order. The may result from mass transfer limitation and shear damage caused by agitation during aggregation. The rugged aggregate showed a higer viability and albumin secretion rate than the dispersed cells or the other aggregates. This result indicates the possible enhancement of a bioartificial liver's (BAL) performance using primary hepatocytes and the reduction in time to prepare a BAL through optimization of the immobilization time.

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Similarities of Tobacco Mosaic Virus-Induced Hypersensitive Cell Death and Copper-Induced Abiotic Cell Death in Tobacco

  • Oh, Sang-Keun;Cheong, Jong-Joo;Ingyu Hwang;Park, Doil
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.8-13
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    • 1999
  • Hypersensitive cell death of plants during incompatible plant-pathogen interactions is one of the efficient defense mechanisms of plants against pathogen infections. For better understanding of the molecular mechanisms involved in the plant hypersensitive response (HR), TMV-induced biotic plant cell death and CuSO4-induced abiotic plant cell death were compared in terms of expression patterns of ten different defense-related genes as molecular markers. The genes include five pathogenesis-related protein genes, two plant secondary metabolite-associated genes, two oxidative stress-related genes and one wound-inducible gene isolated from tobacco. Northern blot analyses revealed that a same set of defense-related genes was induced during both biotic and abiotic cell death but with different time and magnitude. The expression of defense-related genes in tobacco plants was temporarily coincided with the time of cell death. However, when suspension cell cultures was used to monitor the expression of defense-related genes, different patterns of the gene expression were detected. This result implies that three are common and, in addition, also different branches of signaling pathways leading to the induced expression of defense-related genes in tobacco during the pathogen- and heavy metal-induced cell death.

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Taxol Production in Taxus Cell Cultures : 2. Effects of Sugar Concentration (주목 세포배양에 의한 Taxol 생산:2. 당농도의 영향)

  • 김진훈;박인석
    • KSBB Journal
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    • v.9 no.5
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    • pp.450-456
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    • 1994
  • The effects of sucrose on cell growth and anticancer drug taxol production were investigated in cell suspension cultures of Taxus cuspidata. The cells were cultured at various concentrations of sucrose (20 to $100g/\ell$). The highest specific growth rata was achieved at $40g/\ell$ of sucrose as 0.076 day-1 and the highest final cell density, 34 g DCW/$\ell$ after 25 days of culture, was obtained at $60g/\ell$ of sucrose. The cell yield(Yx/s) was found to be 0.55g cell/g sucrose and doubling time (Td) was 9.12 day. High concentration of sucrose (80, $100g/\ell$) and the addition of osmoticum enhanced the production of taxol significantly. The maximum taxol production was $1.36mg/\ell$ at $80g/\ell$ of sucrose, which was 0.01% as a specific content.

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