• Title/Summary/Keyword: Cell suspension cultures

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Production of hGM-CSF by transformed rice cell suspension culture

  • Sin, Yun-Ji;Hong, Sin-Yeong;Kim, Nan-Seon;Kim, Yeong-Suk;Lee, Jae-Hwa;Gwon, Tae-Ho;Yang, Mun-Sik
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.206-209
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    • 2001
  • Recombinant human GM -CSF was expressed and secreted from transgenic rice cell suspension cultures in its biologically active form. This was accomplished by transforming rice callus tissues with an expression vector, pMYN44. containing the hGM -CSF cDNA. Regulated expression and secretion of hGM -CSF from this vector achieved using the promoter, signal peptide, and terminator from a rice alfa-amylase gene Amy3D. The Amy3D gene is expressed in response to sugar deprivation. The recombinant hGM -CSF was expressed from the transgenic rice cell culture on the sugar-free medium as a yield of about 110 mg/L in the culture filtrate, which was determined by ELISA. Biological activity of hGM-CSF was confirmed by measuring the proliferation of the hGM -CSF dependent TF -1 cells.(This work was supported by a grant from the NRL program of the Korean Ministry of Science and Technology. Shin, Y.- J.. Lee. J.-H and Kwon, T.-H. have been supported by BK21 program from the Korean Ministry of Education)

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A Kinetic Modeling for the Dynamics of Hybridoma Cells in Suspension Culture (현탁배양 하이브리도마 세포의 속도론적 모델링)

  • 정연호;박현규최정우
    • KSBB Journal
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    • v.11 no.3
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    • pp.276-287
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    • 1996
  • Batch suspension cultures of hybridoma cell were performed with various initial glutamine concentrations to investigate the effects of glutamine on cell growth and death, monoclonal antibody production, glucose and glutamine consumption, and the production of lactate and ammonium ion. An mathematical kinetic model was formulated to describe the kinetics of cell growth, the consumption of nutrients (glucose and glutamine), and the production of monoclonal antibody and waste metabolites (lactate and ammonium ion) based on experimental data. An equation for the specific growth rate was developed such that superimposed Monod equation in glucose and glutamine, with non-competitive type inhibition relations in ammonium ion and lactate. The inhibition constant for lactate was inversely proportional to the lactate concentration. The specific death rate was considered to be a function of glucose, glutamine, ammonium ion and lactate concentration.

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Enhancement of eurycomanone biosynthesis in cell culture of longjack (Eurycoma longifolia) by elicitor treatment

  • Nhan, Nguyen Huu;Loc, Nguyen Hoang
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.340-346
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    • 2018
  • In this study, the effect of elicitors such as yeast extract (YE), methyl jasmonate (MeJA) and salicylic acid (SA) on the accumulation of eurycomanone in Eurycoma longifolia cell cultures were investigated. Suspension cells of E. longifolia was cultured in Murashige and Skoog (MS) medium supplemented with 30 g/L sucrose, 1.25 mg/L naphthaleneacetic acid (NAA) and 1 mg/L kinetin at a shaking speed of 120 rpm. Elicitors were added in the culture at different concentrations and times to stimulate eurycomanone accumulation in the Eurycoma longifolia cells. Eurycomanone content was determined by HPLC with a C18 column, flow rate of 0.8 mL/min, run time of 17.5 min, and a detector wavelength of 254 nm. The stationary phase was silica gel and the mobile phase was acetonitrile: $H_2O$. Non-elicited cells were used as the control. The study showed the effect of different elicitor concentrations, YE at 200 mg/L, MeJA at $20{\mu}M$ and SA at $20{\mu}M$ stimulated high production of eurycomanone. In which, treatment of $20{\mu}M$ MeJA after 4 days of culture resulted in the highest accumulation of this compound (17.36 mg/g dry weight), approximately 10-fold higher than that of untreated cells (1.70 mg/g dry weight).

Antitumor Activity of Cell Suspension Culture of Green Tea Seed (Camellia sinensis L.)

  • Choi, Jae-Hoon;Yoon, Sang-Kun;Lee, Keyong-Ho;Seo, Min-Soo;Kim, Doo-Hwan;Hong, Seung-Beom;Kim, Ji-Yeon;Paik, Hyun-Dong;Kim, Chang-Han
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.396-401
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    • 2006
  • The objective of this study was to investigate the antitumor activity of suspension cultures of tea callus cells grown in the presence of different concentrations of the growth regulator 2,4-dichlorophenoxy acetic acid (2,4-D) with or without light irradiation. The methanol and ethanol extracts of precipitated cells (MEP, EEP) exhibited stronger inhibitory effects on the growth of tumor cell lines than the water extract of precipitated cells (WEP) or the supernatant Compared to culture under dark conditions, exposure to light irradiation led to significantly higher antitumor activity. The MEP from light irradiated cells at $250{\mu}g/mL$ with 2.0mg/L 2,4-D displayed more than 64% growth inhibition of HEP-2 cells, whereas normal cells showed less than 25% growth inhibition. The some fractions of MEP obtained from Diaion HP-20 column chromatography displayed the majority of inhibitory activity against the HEP-2 cell line. These results show that 2,4-D, and light stimulated the synthesis of antitumor compounds.

Cryopreservation of Suspension Cultures of Camptotheca acuminata

  • Kim, Sung-Hye;Byun, Sang-Yo
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.327-332
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    • 2000
  • Supended cells of Camptotheca acuminata were observed to lose their ability to synthesize camptothecin and its derivatives as a result of repeated cultures. Accordingly, the maintenance of high-yield cells by cryopreservation was sudied to overcome this stability problem, and various factors involved were optimized. Pregrowing the cells in 8% myoinositol for 4 days was found to be the most effective in improving survival. The highest survival was obtained when the pregrown cells were cryoprotected with a mixture of 10% DMSO, 0.6M mannitol, and 10% glycerol. When the cryopreserved cells were maintained in a freezer at $-70^{\circ}C$, 94% survival was obtained after 4 months. The survivals after 5 and 8 months of storage decreased to 52% and 45%, respectively. No loss of biosynthetic capacility of camptothecin was observed after short to medium term cryopreservation of C. acuminata.

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Elicitation of Camptothecin Production in Cell Cultures of Camptotheca acuminata

  • Song, Seung-Hoon;Byun, Sang-Yo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.91-95
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    • 1998
  • Camptothecin productoin was increased with elicitors, methyl jasmonate, jasmonic acid, yeast extract elicitor, and ferulic acid in suspension cultures of Camptotheca acuminata. Jasmonic acid was found to be the most efficient elicitor. Camptothecin production increased 11 times by using the optimum dosing concentration of jasmonic acid which was 50 ${\mu}$M. The kinetics of camptothecin accumulation in response to the treatment with jasmocin acid showed that the comptothecin accumulation reached the maximum value at 4 days after jasmonic acid dosing and then a rapid decrease in camptothecin accumulation was observed.

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Extracellular compounds can enhance development of carrot somatic embryos (세포외 분비물질을 이용한 체세포배 생산성의 향상)

  • 정욱진
    • KSBB Journal
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    • v.11 no.1
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    • pp.107-114
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    • 1996
  • The enhancing effect of excreted cell factors on the production of somatic embryos from suspension cultures of Daucus carota was studied as a function of factors including molecular size, harvesting time, injection period, and concentration of the extracellular compounds. The production of late-stage embryos was increased up to 1, 500 embryos/ml compared with control cultures when high molecular size and extracellular factors, extracted from newly established embryo culture at early stationary phase, were added at the starting time. The stimulating effect on the production of somatic embryos can be attributed to the presence of high molecular size(> 10 kDa) compounds.

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참당귀(Angelica gigas Nakai) 현탁세포배양에서 면역증강성 다당의 생산 증진

  • Jo, Yun-Jeong;Kim, Ik-Hwan;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.479-482
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    • 2001
  • Suspension cells of Angelica gigas Nakai was cultivated to produce extracellular polysaccharide(ECP) as immunostimulating agents. In this study, effects of pectinolytic enzymes such as pectinase. Pectinex on cell growth and ECPs production were investigated in suspension cultures. Optimum concentration of pectinase for ECP production ,vas found to be 0.1 U/L. at which ECP production was increased 1.39 fold (0.97 g/L) compared to control culture.

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Instability of Anthocyanin Accumulation in Vitis vinifera L. var. Gamay Freaux Suspension Cultures

  • Qu Junge;Zhang Wei;Yu Xingju;Jin Meifang
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.155-161
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    • 2005
  • The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, in Vitis vinifera cell cultures. Therefore, four cell line suspensions of Vitis vinifera L. var. Gamay Freaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of $2.73\;\pm\;0.15,\;1.45\;\pm\;0.04,\;0.7\;\pm\;0.024\;and\;0.27\;\pm\;0.04$CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and $84\%$ for V. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be $9.7\%$, ranging from 4 to $17\%$. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities to L-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.

Suspension Culture of Gardenia jasminoides Ellis Cell for Production of Yellow Pigment

  • Kim, Sang-Hwa;Park, Young-Goo;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.2
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    • pp.142-149
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    • 1991
  • Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.

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