• Title/Summary/Keyword: cell suspension

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High Frequency Plant Regeneration in Embryogenic Cell Suspension Cultures of Cucumber (오이 배발생세포의 현탁배양을 통한 고빈도 식물체 재분화)

  • 정원중;우제욱;박효근;최관삼;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.289-291
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    • 1999
  • Hypocotyl explants from 7 days old seedlings of one $F_1$ hybrid cultivar and two pure lines of cucumber formed embryogenic calli at frequencies of up to 8% when cultured on Murashige and Skoog medium (MS) supplemented with 1 mg/L 2,4-D for 3 weeks. Embryogenic calli gave rise to somatic embryos. When slices of somatic embryos were cultured on the same medium for 4 weeks, they formed embryogenic calli. Embryogenic cell suspension cultures were established with embryogenic calli in MS liquid medium with 1 mg/L 2,4-D. Embryogenic potential of cell suspension cultures was maintained by subculturing every seven days. When the level of 2,4-D in the medium was lowered to 0.2 mg/L by diluting with liquid MS basal medium, embryogenic cell suspension cultures underwent development into numerous somatic embryos. When plated onto MS basal medium, over 95% of somatic embryos developed into plantlets. Plantlets were transplanted to potting soil and grown to maturity.

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Pulmonary passage of canine adipose tissue-derived mesenchymal stem cells through intravenous transplantation in mouse model

  • Jaeyeon Kwon;Mu-Young Kim;Soojung Lee;Jeongik Lee;Hun-Young Yoon
    • Journal of Veterinary Science
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    • v.25 no.3
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    • pp.36.1-36.15
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    • 2024
  • Importance: The intravenous administration of adipose tissue-derived mesenchymal stem cells (AdMSCs) in veterinary medicine is an attractive treatment option. On the other hand, it can result in severe complications, including pulmonary thromboembolism (PTE). Objective: The present study assessed the occurrence of PTE after the intravenous infusion of canine AdMSCs (cAdMSCs) into experimental animals. Methods: Five-week-old male BALB/c hairless mice were categorized into groups labeled A to G. In the control group (A), fluorescently stained 2×106 cAdMSCs were diluted in 200 µL of suspension and injected into the tail vein as a single bolus. The remaining groups included the following: group B with 5×106 cells, group C with 3×106 cells, group D with 1×106 cells, group E with 1×106 cells injected twice with a one-day interval, group F with 2×106 cells in 100 µL of suspension, and group G with 2×106 cells in 300 µL of suspension. Results: Group D achieved a 100% survival rate, while none of the subjects in groups B and C survived (p = 0.002). Blood tests revealed a tendency for the D-dimer levels to increase as the cell dose increased (p = 0.006). The platelet count was higher in the low cell concentration groups and lower in the high cell concentration groups (p = 0.028). A histological examination revealed PTE in most deceased subjects (96.30%). Conclusions and Relevance: PTE was verified, and various variables were identified as potential contributing factors, including the cell dose, injection frequency, and suspension volume.

Plant Regeneration from Embryogenic Suspension Culture of Orchardgrass (Dactylis glomerata L.) (오차드그래스의 현탁배양으로부터 부정배 형성과 식물체 재분화)

  • 이효신;권용삼;이병현;원성혜;김기용;조진기
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.20 no.1
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    • pp.7-12
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    • 2000
  • This study was carried out to improve the ability of embryo formation and the efficiency of plant regeneration from suspension cultured cells of seed derived calli of orchardgrass (Dactylis glomerata L.). The frequency of formation of round cell and cell colonies was highest at 50 days after suspension culture in $N_6$ medium supplemented with $4\;g/{\ell}$ casein hydrolysate (CH), $20\;g/{\ell}$ sucrose and $30\;g/{\ell}$ sorbitol. The highest frequency of plant regeneration and somatic embryo formation was obtained from suspension cultured cells of 60 days. Addition of CH ($4\;g/{\ell}$) in suspension culture medium gave the highest frequency of embryo formation (39.6%) and plant regeneration (73.0%).

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Production of Recombinant Polyhedra Containing Cry1Ac Fusion Protein in Insect Cell Lines

  • Kim, Jae-Su;Choi, Jae-Young;Roh, Jong-Yul;Lee, Han-Young;Jang, Seung-Sik;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.739-744
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    • 2007
  • Insect cell lines and the control of infection for obtaining the maximum amount of polyhedrin-Cry1Ac-polyhedrin fusion protein from Bactrus in monolayer and suspension culture systems were tested. Growth rates of the Trichoplusia ni(High-Five) cell line in both culture systems were better than the other insect cell lines, Spodoptera frugiferda(Sf-9, Sf-21), Trichoplusia ni(Tn5), and Spodoptera exigua(Se301). The expression of the fusion protein in a monolayer culture showed that Se301 cells were 2.3-4.8 times more productive on a per cell basis than the other cell lines. However, in suspension culture, only High-Five cells were productive. High-Five cells infected with Bactrus at a multiplicity of infection(MOI) of 5 and a cell density of $3.0{\times}10^5$ cells per ml were more productive than the other infection condition in a suspension culture suitable for a large-scale production of baculovirus. In conclusion, for the large-scale production of Bactrus in vitro, High-Five cells showing good growth and high productivity are suitable.

Characterization of Cell Growth and Camptothecin Production in Cell Cultures of Camptotheca acuminata

  • Song, Seung-Hoon;Byun, Sang-Yo
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.631-638
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    • 1998
  • Studies were made to elucidate the cell growth and the production of camptothecin and its derivatives in cell cultures of Camptotheca acuminata. High resolution HPLC chromatograms to analyze camptothecin and 10-hydroxycamptothecin in lactone and carboxylate forms were obtained with a fluorescence detector. Calli inductions were optimized with the young stem of explant on Schenk and Hildebrandt (SH) medium supplemented with 5 mg/l $\alpha$-naphthaleneacetic acid (NAA), 0.2 mg/l 6-benzylamino purine (BAP), 2.0% sucrose, and 0.5% agar. The hybrid medium, a mixture of SH and Murashige and Skoog (MS) salts, was developed for homogeneous suspension cultures without large cell aggregates. The optimum phytohormone concentrations for successful suspension cultures were 1.0mg/l of 2,4-D and 0.5 mg/l of kinetin. The highest growth in suspension cultures was observed when 49.7% (w/w) of the cells was composed of small aggregates which were below 0.1 mm in diameter. Time course changes of cell growth and camptothecin production showed that camptothecin accumulation was started at the end of the growth phase and the maximum content was obtained 10 days after inoculation. Yeast extract elicitor increased camptothecin accumulation 4 times. Methyl jasmonate and jasmonic acid also increased camptothecin production 6 and 11 times, respectively.

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Effects of Sucrose level and Nitrogen Source on Fresh Weight and Anthocyanin Production in Cell Suspension Culture of 'Sheridan' Grape (Vitis spp.)

  • Kim, Seung-Heui;Kim, Seon-Kyu
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.23-27
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    • 2002
  • To establish an in vitro mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of nitrogen source and sucrose on fresh weight and anthocyanin production in cell suspension culture of 'Sheridan' grape level were studied. When the medium was devoid of $NO_3^-$, cell fresh weight was either remained stable (1% sucrose) or slightly decreased with culture time (2,3, and 4% sucrose). When $NH_4^-$ was lacking, 3% sucrose was most favorable for cell growth. When $NH_4^-$ was supplied as N source, the anthocyanin content of 2% sucrose containing medium was maintained 2 times higher than other levels till day 8 in culture, then that of 3 and 4% sucrose which peaked at day 12 thereafter. The anthocyanin content was low than $NO_3^-$-free media. Total anthocyanin content in $NH_4^-$-free medium was just about a half of that of $NH_4^+$ medium. Anthocyanin production of 2% sucrose in $NH_4^+$ medium was maintained about 3-fold till day 8, then decreased thereafter. In $NH_4^+$ medium, pH decreased gradually with final pH of 3.5 to 4.0, while pH in $NH_4^+$-free medium increased with final pH of 6.5 to 7.5.

Growth Prpmotion of Taxus brevifolia Cell Suspension Culture Using Conditioned Medium

  • Kim, Myung-Hwan;Chun, Su-Mwan;Kim, Dong-Il
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.5
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    • pp.350-354
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    • 2000
  • The growth promotion of a Taxus brevifolia cell suspension culture was investigated using conditioning factors. The conditioning factors produced and secreted from cultured cells usually stimulate cell division and the production of secondary metabolites. Therefore, the effective incubation time for the optimal secretion of conditioning factors was firstly determined for the promotion of cell growth. Conditioned media obtained by cultivating for 2 and 5 days showed the promotion of initial cell growth during the early cell growth period. However, the positive effect of the conditioning factors on the initial cell growth did not continue because of the depletion of the medium nutrients. Accordingly, the addition of a carbon source to the conditioned medium prolonged the positive effect on the cell growth. The addition of sucrose to the conditioned medium resulted in the maximum cell density being reached 4 days earlier compared to the control group and an increased substrate yield.

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Critical Suspension Condition of Particles in a Shaking Vessel of Solid-Liquid System (고-액계 진동교반에서 입자의 부유화 한계조건)

  • Lee, Young-Sei;Kim, Moon-Gab;Kato, Yoshihito
    • Journal of the Korean Society of Industry Convergence
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    • v.2 no.2
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    • pp.73-81
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    • 1999
  • Shake mixing has been widely used in cell culture. The mixing performance for shake mixing, however, has not been reported quantitatively. The critical circulating frequency and the power consumption for complete suspension of particles, based on the definition of Zwietering, were measured in a shaking vessel containing a solid-liquid system. The critical suspension frequency was correlated by the equation from Baldi's particle suspension model modified with the physical properties of the particles. Critical suspension frequency was correlated as following ; $$N_{JS}={\frac{0.58\;d{_p}^{0.06}(g{\Delta}{\rho}/{\rho}_L)^{0.004}X^{0.03}}{D^{0.35}d^{0.17}{\upsilon}^{0.04}}}$$ The power consumption at the critical suspension condition in the shaking vessel was less than that in an agitated vessel with impeller.

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Anthraquinones from Cell Suspension Culture of Morinda elliptica

  • Jasril, Jasril;Lajis, N.H.;Abdullah, M.A.;Ismail, N.H.;Ali, A.M.;Marziah, M.;Ariff, A.B.;Kitajima, M.;Takayama, H.;Aimi, N.
    • Natural Product Sciences
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    • v.6 no.1
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    • pp.40-43
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    • 2000
  • The chemical investigation on the cell suspension culture of Morinda elliptica L. yielded eight anthraquinones, two of which, anthragallol-1,2-dimethyl ether (3) and purpurin-1-methyl ether (4), have not been isolated from the original plant. Other compounds isolated include nordamnacanthal (1), alizarin-1-methyl ether (2), rubiadin (5), soranjidiol (6), $lucidin-{\omega}-methyl$ ether (7), and morindone (8). The structures of anthraquinones were established based on spectral studies.

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Effects of $Ca^{2+}$ and Polyamine on Callose Contents in Carrot Suspension Cultured Cells (당근 현탁배양세포에서 $Ca^{2+}$과 Polyamine이 Callose 함량에 미치는 영향)

  • 강영희
    • Journal of Plant Biology
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    • v.32 no.4
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    • pp.215-226
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    • 1989
  • The effects of Ca2+ on polyamines on callose contents of carrot suspension cultured cells were studied. The regeneration process of the cell wall of carrot protoplast observed through the electron microscope. Treatment of the carrot suspension cultured cells with Ca2+ and polyamines resulted in considerable increase on callose contents at 0.1 mM of Ca2+ and polyamines, particulary spermidine. Poly-L-lysine and poly-L-ornithine increased about 30% and 100% of callose contents than that of the control respectively, whereas verapamil and flunarizine markedly decreased the callose contents. These effects of Ca2+ of free ion rather than as Ca2+-calmodulin complex. During the cultivation of the protoplast, the regeneration of the cell wall was somewhat observed on the 4th day, however, it was inhibited by verapamil. These results suggested that the promotive action of Ca2+ and polyamines were manifested in the callose contents and the regeneraton of the cell wall.

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