• Title/Summary/Keyword: Cell Transformation

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Efficient Plantlet Regeneration via Callus Formation from Leaf Segment of Lilium Oriental Hybrid 'Casa Blanca'

  • Kim Mi-Sun;Jeon Jae-Heung;Youm Jung-Won;Kim Jae-Hyun;Lee Byung-Chan;Kang Won-Jin;Kim Hyun-Soon;Joung Hyouk
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.129-134
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    • 2005
  • Callus induction from a leaf explant has been achieved in Lilium Oriental hybrid 'Casa Blanca'. The highest frequency of callus induction was obtained on MS medium supplemented with 0.5 mg/L BA and 2.0 mg/L NAA after 2 months of culture. The cultures maintained continuously without change in color and type of callus when they cultured in the dark. Plantlet regeneration with a high frequency was achieved from induced calli on the same medium. A number of shoots are formed from one cluster of callus, and bulblets developed into intact plantlets after transfer to hormone-free MS medium. No phenotypic variations were observed among regenerants. Enhancement in plantlet regeneration via callus formation would be expected to facilitate the efficiency of transformation of this Oriental hybrid 'Casa Blanca'.

Usage of Human Mesenchymal Stem Cells in Cell-based Therapy: Advantages and Disadvantages

  • Kim, Hee Jung;Park, Jeong-Soo
    • Development and Reproduction
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    • v.21 no.1
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    • pp.1-10
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    • 2017
  • The use of human mesenchymal stem cells (hMSCs) in cell-based therapy has attracted extensive interest in the field of regenerative medicine, and it shows applications to numerous incurable diseases. hMSCs show several superior properties for therapeutic use compared to other types of stem cells. Different cell types are discussed in terms of their advantages and disadvantages, with focus on the characteristics of hMSCs. hMSCs can proliferate readily and produce differentiated cells that can substitute for the targeted affected tissue. To maximize the therapeutic effects of hMSCs, a substantial number of these cells are essential, requiring extensive ex vivo cell expansion. However, hMSCs have a limited lifespan in an in vitro culture condition. The senescence of hMSCs is a double-edged sword from the viewpoint of clinical applications. Although their limited cell proliferation potency protects them from malignant transformation after transplantation, senescence can alter various cell functions including proliferation, differentiation, and migration, that are essential for their therapeutic efficacy. Numerous trials to overcome the limited lifespan of mesenchymal stem cells are discussed.

Production of Useful Proteins by Plant Cell Culture

  • Kwon, Tae-Ho;Kim, Dae-Hyun;Jang, Yong-Suk;Yang, Moon-Sik
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.45-49
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    • 1999
  • Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

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Development of an Agrobacterium-mediated Transient Expression System for Intact Leaves of Chili Pepper (Agrobacterium을 이용한 고추의 Transient Expression 시스템)

  • Seong, Eun-Soo;Joung, Young-Hee;Choi, Doil
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.185-190
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    • 2004
  • We established a transient gene expression system in chili pepper leaves based on Agrobacterium-mediated transformation of GUS gene. For the best GUS transient expression, two step culture system was adopted. When the Agrobacterium tumefaciens cell density of pre-culture was $A_{600nm}$ 0.3, the cells were harvested and diluted to $A_{600nm}$ 0.8 with virulence induction medium after cell harvested. The addition of acetosyringone (200 $\mu$M) in virulence induction step was a key factor for successful transient expression. Additionally, Younger leaves showed more effective transient expression than older leaves. Temporally, the strongest intensity of GUS expression was detected at 2 days after infiltration. These results demonstrate that Agrobacterium-mediated transient expression can be used for a simple in vivo assays of plant promoters, transcription factors and furthermore provide efficient protocol for chili pepper transformation.

Biodegradation Study of Gasoline Oxygenates by Butane-Utilizing Microorganisms (부탄 분해 미생물을 이용한 휘발유 첨가제의 분해특성)

  • 장순웅
    • Journal of Soil and Groundwater Environment
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    • v.8 no.1
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    • pp.27-34
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    • 2003
  • In this study, potential degradation of MTBE and other gasoline oxygenates by pure culture ENV425 and mixed culture isolated from gasoline contaminated soil using butane as the sources of carbon and energy was examined and compared. Butane monooxygenases(BMO) of butane-grown ENV425 and mixed culture generated 1-butanol as a major metabolite of butane oxidation and addition of acetylene, specific inhibitor of monooxygenase, inhibited both butane oxidation and 1-butanol production. The results described in this study suggest that alkanes including propane, pentane, and butane are effectively utilized as a growth substrate to oxidize MTBE cometabolically. And also BTEX compounds could be the potential substrate of the MTBE cometabolism. Cell density also affected on the MTBE degradation and transformation capacity(Tc). Increasing cell density caused increasing MTBE degradation but decreased transformation capacity. Other result demonstrated that MTBE and other gasoline oxygenates, ETBE and TAME, were degraded by butane-grown microorganism.

The Segmentation and the Extraction of Precise Plane Equation of Building Roof Plane using 3D Hough Transformation of LiDAR Data (LiDAR 데이터의 3D Hough 변환을 이용한 건물 지붕 평면의 세그멘테이션 및 정밀 평면방정식 추출)

  • Lee, Young-Jin;Oh, Jae-Hong;Shin, Sung-Woong;Cho, Woo-Sug
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.26 no.5
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    • pp.505-512
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    • 2008
  • The 3D Hough transformation is the one of the most powerful and popular algorithm for extracting plane parameters from LiDAR data. However, there are some problems when extracting building roof plane using 3D Hough transformation. This paper explains possible problems and solution for extracting roof plane. The algorithm defines peak plane, exact plane, and LESS plane for extracting accurate plane parameters in the accumulator of the 3D Hough transformation. The peak plane is the plane which is represented by peak in the accumulator. The exact plane is the plane which is represented by the accumulator cell which is closest to the actual plane. The LESS plane can be calculated from all LiDAR points in the exact plane by using least-square adjustment. Test results show that proposed algorithm can extracts building roof plane very accurately.

Chemical Transformation of Human Keratinocytes by 2,3,7,8-Tetrachlorodibenxo-$\rho$-dioxin

  • Kang, Mi-Kyung;Choi, Young-Sill;Ryeom, Tai-Kyung;Eom, Mi-Ok;Park, Mi-Sun;Jee, Seung-Won;Kim, Kang-Ryune;Kim, Ok-Hee;Kang, Ho-Il
    • Environmental Mutagens and Carcinogens
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    • v.26 no.3
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    • pp.69-76
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    • 2006
  • 2,3,7,8-Tetrachlorodibenzo-$\rho$-dioxin(TCDD) is a ubiquitous, persistent environmental contaminant and the most powerful carcinogen categorized by IARC. Although the mechanism of carcinogenesis by TCDD is poorly understood, several studies have shown that the skin is one of target organs far TCDD. In this study, we investigated the neoplastic transformation of human keratinocyte-derived cell line, HaCaT, by chemical transformation method using N-methyl-N'-nitro-N-nitrorsoguanidine(MNNG) and TCDD. We found that subsequent exposure to TCDD for 3 weeks after initial exposure to MNNG markedly induced transformed cells. It was suggested that TCDD can act as a potent promoter in HaCaT cells. Furthermore, these transformed cells showed morphological alternations in soft agar and increased telomerase activity. Therefore, the TCDD treatment of HaCaT cells by initiated with MNNG could promote neoplastic transformation without stimulation by exogenous growth factors. As a result, TCDD had a strong potency as a promoter in nontumorigenic immortalized human epidermal keratinocytes.

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Heterologous Transformation of Saccharomyces cerevisiae by Glucoamylase Gene of Saccharomyces diastaticus (Saccharomyces diastaticus Glucoamylase Gene에 의한 Saccharomyces cerevisiae의 Transformation)

  • Kim, Young-Ho;Jun, Do-Youn;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.489-493
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    • 1988
  • To obtain a new yeast strain that is able to efficiently produce ethanol from starch, the glucoamylase gene of Saccharomyces diastaticus was transformed into S. cerevisiae without a cloning vector. The competent cells of S. cerevisiae, induced by the treatment of Li$_2$SO$_4$, were transformed with the partial BamHI-digests of chromosomal DNA of S. diastaticus, and the transformants were selected by their abilities to utilize and ferment starch. The transformants, which appeared at a frequency of 8.5$\times$10$^{-7}$, were able to withstand up to 800 ppm of copper sulfate like the recipient and retained the phenotypic expression of the recipient with the exception of the acquisition of STA gene and MAL gene, as regards fermentation of carbohydrates. The enzymatic properties of glucoamylases produced by transformants were very similar to those produced by S. diastaticus as based on optimium pH and temperature.

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Experimental Studies on the Anti-tumor and the Immunomodulatory Effects of Chungsimbohyeltang (청심보혈탕(淸心補血湯)의 항암(抗癌) 및 면역증강효과(免疫增强效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Park, Se-Ki;Ryu, Bong-Ha;Park, Dong-Won;Ryu, Ki-Won
    • The Journal of Internal Korean Medicine
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    • v.19 no.1
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    • pp.221-246
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    • 1998
  • The aim of this experimental study was to evaluate the anti-tumor effects of Chungsimbohyeltang through investigation about viability of tumor cell by MTT assay, survival period of mice transplanted with L-1210 cells, growth inhibition on the tumor cell, body weight variation in mice transplanted with sarcoma-180 cells and its immunomodulatory effects through the investigation on delayed type hyper-sensitivity, the hemagglutinin and hemolysin titers for humoral immune response, the appearance of rosette forming cells for cell-mediated immune response, the natural killer cell activity, the transformation of lymphocyte, the productivity of Interleukin-2 and phagocytic index K was performed in immune-depressed ICR mice induced by methotrexate treatments. The results were as follows ; 1. $IC_{50}$ of Chungsimbohyeltang treated group was 5.85mg/ml in SNU-C4 cell, 1.38mg/ml in SNU-396 cell, 0.21mg/ml in SNU-1 cell, so it had low anti-tumor activity. 2. The both groups of Chungsimbohyeltang extract 10mg/kg and Chungsimbohyeltang extract 20mg/kg had no toxicity and the group of Chungsimbohyeltang 20mg/kg which was shown 120% in ILS had the effect of life prolongation in mice transplanted with L-1210 cells. 3. In the growth inhibition on the tumor cells, only the group of Chungsimbohyeltang extract 20mg/kg was noted and in the weight variation in mice transplanted with sarcoma-180 cells, both groups of Chungsimbohyeltang extract had a significant effect. 4. In the delayed type hypersensitivity and appearance of rosette forming cells, both groups of Chungsimbohyeltang extract didn't have any significant effect. 5. The hemagglutinin titers was slightly increased with no significance, and the hemolysin titers was significantly increased in the only group of Chungsimbohyeltang extract 20mg/kg. 6. The natural killer cell activity of the Chungsimbohyeltang extract groups was significantly increased in the ratio of 100:1 of effector and target cells, but it was not significantly increased in the ratio of 50:1, 10:1. 7. The transformation of lymphocyte and the productivity of Interleukin-2 were increased significantly and in dose-dependent manner in both group of Chungsimbohyeltang extract. 8. The phagocytic effect of macropage was significantly increased in both groups of Chungsimbohyeltang extract. Considering the results above, we could conclude that Chungsimbohyeltang have an indirect anti-tumor effect through the modulation of immunme response, although it had not toxicity on the tumor cell it self.

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Transgenic Plants of Easter Lily (Lilium longiflorum) with Phosphinothricin Resistance

  • Ahn, Byung Joon;Joung, Young Hee;Kamo, Kathryn K.
    • Journal of Plant Biotechnology
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    • v.6 no.1
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    • pp.9-13
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    • 2004
  • Transient uidA expression was used to optimize parameters required for biolistic transformation of suspension cells of Easter lily, Lilium longiflourm. Maximum uidA expression occurred following bombardment with gold particles as compared to tungsten. A 3hr pre-treatment of suspension cells with 0.125M osmoticum resulted in a 1.5X increase in uidA expression. A helium pressure of 1550 psi combined with a particle travelling distance of 6cm resulted in maximum uidA expression as compared to either 1100, 1200, or 1800 psi. Transient transformation resulted in up to 493 uidA expressing cells/Petri plate. For stable transformation suspension cells of Lilium longiflorum, were co-bombarded with plasmid DNA containing cucumber mosaic virus (CMV) replicase under the rice actin (Act1) promoter and either the bar or PAT genes under the cauliflower mosaic virus (CaMV 355) promoter. Ten regenerated plants contained the transgene as analyzed by PCR, and two of the ten plants were confirmed to contain the transgene by Southern hybridization. The two transgenic plants were independent transformants, one containing the bar gene and the other both the CMV replicase and bar genes. Plants were sprayed at the rosette stage and found to be resistant to 1000 mg/L of phosphinothricin (Trade name-Ignite) indicating expression of the bar gene throughout the leaves when bar was under control of the CaMV 35S promoter.