• Title/Summary/Keyword: suspension-cultured-cells

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The Effects of Ammonium Citrate and Ammonium Succinate on the Growth of Cells and Nitrogen Absorption in Korean Ginseng Suspension Cultures (고려인삼(Panax ginseng C.A. Meyer) 배양세포의 생육 및 질소 흡수에 미치는 Ammonium Citrate와 Ammonium Succinate의 영향)

  • 김홍성;김명원;소상섭;강영희
    • Journal of Plant Biology
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    • v.24 no.1
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    • pp.13-19
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    • 1981
  • In order to investigate the effects of ammonium citrate and ammonium succinate on the growth and absorption of nitrogen compounds supplied in the medium, Korean ginseng (Panax ginseng C. A. Meyer) calli were suspension cultured in MS medium with various concentrations of ammonium citrate and ammonium succinate. When Korean ginseng calli were cultured with 10 mM ammonium citrate, 10 mM ammonium succinate, and 10 mM ammonium nitrate (control) in MS media as the nitrogen sources, the growth, $NO_3$-N absorption and total nitrogen content of the Korean ginseng cells were greatest in the ammonium citrate and ammonium succinate concentrations. When Korean ginseng calli were cultured with 5 mM ammonium citrate and 5 mM ammonium succinate, the growth and nitrogen content were superior to those of the control: however, $NO_3$-N and $NH_4$-N absorptions were similar to those of the control. In conclusion, the 10 mM ammonium citrate and 10 mM ammonium succinate may be better able to facilitate the growth and $NO_3$

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Effect of Cadmium on Somatic Embryogenesis from Cell Culture of Daucus carota L. (당근(Daucus carota L.)의 현탁배양을 통한 체세포배 발생에 미치는 카드뮴의 영향)

  • 조덕이;신은경;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.227-232
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    • 2000
  • This study was carried out to elucidate the effect of cadmium on somatic embryogenesis and plant regeneration from cultured cells of Daucus carota L. Embryogenic calli were induced from cotyledon explants of carrot seedlings cultured on MS solid medium supplemente with 1 mg/L 2,4-D Embryogenic cells proliferated on medium supplemented with 1 mg/L 2,4-D were also cultured in liquid MS medium containing various concentrations (50, 100, 200, 500, 1000 $\mu$M) of cadmium for one week and then transferred to MS basal medium. Somatic embryogenesis occurred in suspension culture treated with 50 $\mu$M and 100 $\mu$M cadmium or untreated with cadmium. When cadmium was treated in suspension culture, production of two and four cotyledonary somatic embryos was reduced, but that of three cotyledonary somatic embryo was increased. Two cotyledonary embryos showed higher regeneration frequency than abnormal somatic embryo with one, three and four cotyledon. Regardless of cotyledonary variation, germination frequency of somatic embryos treated with cadmium was decreased in compared with that of embryos in basal medium.

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Evaluation of Functional Properties of the Tissue Cultured Wild Ginseng Fermented by Lactobacillus sp. (Lactobacillus sp.균주를 이용한 산삼 배양근 발효물의 기능성 평가)

  • Shin, Eun Ji;Cho, Chang-Won;Kim, Young-Eon;Han, Daeseok;Hong, Hee-Do;Rhee, Young Kyoung
    • Journal of the Korean Society of Food Culture
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    • v.27 no.6
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    • pp.743-750
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    • 2012
  • A tissue cultured wild ginseng (TCWG) suspension was inoculated with lactic acid bacteria and fermented to improve the functionality of TCWG. The utilization of TCWG was increased directly using the freeze-dried powder. The optimal ratio of TCWG powder and water for fermentation was 1:19 (5%), which was selected by measuring the fluidity and viable cell count according to concentration. The effects on ADH activation and immune cell activation by each ferments with 10 kinds of Lactobacillus sp. strains were examined. The ferments with the Lactobacillus casei KFRI 692 strain showed 5.4 times higher ADH activity and 1.3 times higher ALDH activity than the non-fermented TCWG powder (control). The level of NO production and cytotoxicity was also measured by Raw 264.7 cells. The ferment with the Lac. casei KFRI 692 strain showed the highest level of NO production and lower cytotoxicity than the others. Therefore, the Lac. casei KFRI 692 strain was selected as a strain for fermentation of a TCWG suspension to maximize its functionality. To identify the optimal fermentation time of the selected Lac. casei KFRI 692 strain on the 5% TCWG suspension, the viable cell count of lactic acid bacterial and the changes in pH were observed for 72 hours. 24-hrs was found to be the optimal fermentation time. In this way, fermented TCWG with lactic acid bacteria showed higher ADH activation efficacy and immune cell activation than non-fermented TCWG.

Taxol Production in Taxus sap. Cell Cultures 1. Studies on Taxol Content in Yew Trees and Cultured Plant Cells (주목 세포배양에 의한 Taxoll 생산 1.주목 부위 및 서식 고도별 Taxol 함량 및 세포주 유도에 관한 연구)

  • 강인선;전정욱
    • KSBB Journal
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    • v.9 no.3
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    • pp.299-305
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    • 1994
  • Taxol contents in various parts of 15 years old yew tree were determined. The descending order of taxol content per unit mass was stem bark, root bark, needle and seed. In the seed, that order was seed coat, embryo and endosperm. The total amount of taxol extractable from a 15 years old yew tree was 1.68 gram. This amount was distributed in needle, stem bark, root bark and seeds as 48.0, 23.8, 27.9 and 0.4%, respectively. Altitudinal variation of taxol content was also observed. More taxol was observed in yew trees grown at high altitude over 1000m above sea level. Calli and suspension cultures were induced from various yew trees. The presence of taxo] in cultured cells was established bv HPLC. The taxol content in cultured cells were different according to the source of explants. These results may be useful for the goal of large scale taxol production by cultured yew tree cells.

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Shoot Proliferation and Plant Regeneration from Suspension-Cultured Cells of Dianthus gratianopol (패랭이꽃속 Dianthus gratianopol의 현탁배양세포로부터 Shoot 증식과 식물체 재분화)

  • Kim Joon-Chul
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.301-306
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    • 2005
  • Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

Relationship between Germ Tube Formation, Adherence to Human Buccal Epithelial Cells and Virulence of Candida albicans (Candida albicans의 상피세포에 대한 부착능과 병원성과의 상관관계에 관한 연구)

  • Koh, Choon-Myung
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.407-415
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    • 1986
  • This study investigated whether a correlation exists between environmental physical and biochemical factors and adherence of Candida albicans to human buccal epithelial cells by using normal and UV-irradiated strains. The results were as follows: 1. The percentage of germ tube forming activities of normal Candida albicans was 91.5% and UV-irradiated Candida albicans was 15.0%. The $LD_{50}$ of normal strains in mice were $1.0{\times}10\;cells/ml$, but could not be observed in the UV-irradiated strains even with $1.0{\times}10\;cells/ml$. It demonstrated that the virulence is decreased in the UV-irradiated strain. 2. The adherence of normal Candida albicans to human buccal epithelial cells($166{\pm}29{\sim}207{\pm}17\;cells$/100 epithelial cells) was significantly greater than UV-irradiated Candida albicans($99{\pm}21{\sim}131{\pm}25\;cells$/100 epithelial cells). 3. Candida albicans cultured at $37^{\circ}C$ adhered to buccal epithelial cells($166{\pm}16{\sim}207{\pm}17\;cells$/100 epithelial cells) in greater numbers than cultured at $25^{\circ}C$($80{\pm}15{\sim}143{\pm}22\;cells$/100 epithelial cells). 4. On comparison of the adherence of viable and nonviable(heat-killed) Candida albicans to human buccal epithelial cells, the nonviable Candida albicans demonstrated poorer adherence than viable Candida albicans. 5. Adherence in vitro of Candida albicans to human epithelial cells appeared to be effected by the pH. The adherence ability was maximum increased at pH 7.0($187{\pm}22\;cells$/100 epithelial cells) other than experimental pH. 6. The adherence was proportional to the incubation time and the Candida cell concentration in the suspension. 7. A strong correlation was shown between germ tube forming activity and increased adherence of Candida albicans to human epithelial cells, indicating that germ tube forming activity were responsible for candidal virulence.

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Cloning and Characterization of Homeodomain-Zip Gene, Phc5 in Embryogenic Callus derived from Pimpinella brachycarpa Suspension Cultured Cells (참나물 현탁배양세포 유래 배발생캘러스에서 HD-Zip 유전자, Phc5의 클로닝과 특성)

  • 손수인;김준철
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.121-126
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    • 1999
  • Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

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Protective Effect of Bcl-2 in NS0 Myeloma Cell Culture is Greater in More Stressful Environments

  • Tey, B.T.;Al-Rubeai, M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.564-570
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    • 2005
  • In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.

Production of Green Fluorescent Protein (GFP) from Transgenic Rice Cell Suspension Culture (형질전환된 벼세포배양에서 green fluorescent protein (GFP) 생산)

  • Lee, Jae-Hwa
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.293-297
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    • 2007
  • Green fluorescent protein (GFP) is an attractive reporter for bioprocess monitoring. A fluorescence-based method was developed to quantify GFP levels in transgenic plants and protein extracts. In this study, GFP was produced and secreted from suspension cells derived from transgenic rice. The RAmy3E promoter placed before the GFP gene controlled by sugars such as sucrose. The effects of sucrose concentration on the secretion of GFP and total protein into the medium were investigated in batch suspension culture. It was possible, therefore, to induce the expression of the GFP by removing sucrose from the cultured media or by allowing the rice suspension cells to deplete sucrose catabolically. The dry cell weight (7.06 g/L) and GFP level were detected as highest at 12%, 3% sucrose after 20 day culture, respectively. However secreted GFP fluorescence at the other sucrose concentrations (6%, 12%, 18% and 24%) were a little amount in media.

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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