• Title/Summary/Keyword: stable cells

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In vivo Dendritic Cell Migration Tracking Using Near-infrared (NIR) Imaging (Near-infrared (NIR) 영상기법을 이용한 생체 내 수지상세포의 이동)

  • Lee, Jun-Ho;Jung, Nam-Chul;Lee, Eun Gae;Lim, Dae-Seog
    • KSBB Journal
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    • v.27 no.5
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    • pp.295-300
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    • 2012
  • Matured dendritic cells (DCs) begin migration with their release from the bone marrow (BM) into the blood and subsequent traffic into peripheral lymphoid and non-lymphoid tissues. Throughout this long movement, migrating DCs must apply specialized skills to reach their target destination. Non-invasive in vivo cell-tracking techniques are necessary to advance immune cell-based therapies. In this study, we used a DiD cell-tracking solution for in vivo dendritic cell tracking in naive mice. We tracked DiD (non-invasive fluorescence dye)-labeled mature dendritic cells using the Near Infrared (NIR) imaging system in normal mice. We examined the immunophenotype of DiD-labeled cells compared with non-labelled mature DCs, and obtained time-serial images of NIR-DC trafficking after mouse footpad injection. In conclusion, we confirmed that DiD-labeled DCs migrated into the popliteal lymph node 24 h after the footpad injection. Here, these data suggested that the cell tracking system with the stable fluorescence dye DiD was useful as a cell tracking tool to advance dendritic cell-based immunotherapy.

High-Level Expression of Recombinant Human Interleukin-2 in Chinese Hamster Ovary Cells Using the Expression System Containing Transcription Terminator

  • Kim, Eun-Ju;Kim, Dong-Jun;Hwang, Hye-Yeon;Yoon, Jae-Seung;Yoon, Ye-Up;Baek, Kwang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.810-815
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    • 2004
  • Many biological properties and the clinical potential of human interleukin-2 (hIL-2) draw much attention to its high-level expression in mammalian cells. Recombinant human IL-2 (rhIL-2) was expressed in Chinese hamster ovary (CHO) cells, using the recently developed expression system which confers position-independent expression. Stable CHO cell lines carrying several hundred amplified copies of the rhIL-2 gene were easily obtained and rhIL-2 was expressed at high levels after selection with increasing concentrations of methotrexate. Interestingly, the insertion of the transcription terminator of the human gastrin gene into the downstream region of the gene for rhIL-2 considerably increased rhIL-2 expression. Using the expression system with the transcription terminator, it was possible to get a CHO cell line expressing the rhIL-2 at a very high level, about $11.4\mug/10^6$ cell/day, which is about 6 times higher than that previously reported. The biological activity of the rhIL-2 protein purified from the cell line was also confirmed by the cell proliferation assay.

Enhancement of Erythropoietin Production from Chinese Hamster Ovary(CHO) Cells by Introduction of the Urea Cycle Enzymes, Carbamoyl Phosphate Synthetase I and Ornithine Transcarbamylase

  • Kim, Na-Young;Lee, Yun-Jeong;Kim, Hyung-Jin;Choi, Jung-Ho;Kim, Jung-Kwon;Chang, Kern-Hee;Kim, Jung-Hoe;Kim, Hong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.844-851
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    • 2004
  • Efficient mammalian erythropoietin (EPO)-expression systems are required for therapeutic applications. The accumulation of ammonia is a major problem in the production of recombinant proteins in cultured animal cells. To counter this problem we introduced the first two genes of the urea cycle, carbamoyl phosphate synthetase (CPSI) and ornithine transcarbamylase (OTC), into IBE Chinese Hamster Ovary (CHO) cells by stable transfection. The resulting cell line, CO5, had a higher growth rate and accumulated less ammonia per cell than the parental cell line, IBE. In addition, it produced 2 times more EPO than the parent, and the purified EPO contained a higher proportion of acidic isoforms with approximately 15% more sialic acid.

Selection of Immobilization Material for Stabilization of Bioluminescence from Photobacterium phosphoreum (Bioluminescence 안정성을 위한 Photobacterium phosphoreum의 고정화 물질에 관한 연구)

  • Lee, Eun-Su;Kim, Hyeon-Suk;Jeon, Eok-Han
    • KSBB Journal
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    • v.14 no.4
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    • pp.403-407
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    • 1999
  • Various materials including sodium alginate, k-carragreenan, collagen and polyacrylamide were studied in order to maintain stability of bioluminescence of P. phosphoreum for the purpose of continuos monitoring of toxic subtances. Collagen and polycryamide were shown to be inadequate for immobilization of p. phosphoreum since the bioluminescence decreased when cells were mixed with such materials. In case of k-carrageenan, the bioluminescence was stable when compared with collagen and polyacryamide. However, the k-carrageenan was not suitable for immobilization of p. phosphoreum as cells could not be mixed with the material properly in temperature at which gel formation already occurred. P . phosphoreum must be treated at low temperature below that of gel formation since these are psychrophilic luminescent bacterial. When cells were immobilized on sodium alginate, the bioluminescence was stably maintained for 20 minutes.

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Physiological Characteristics of Immobilized Streptomyces Cells in Continuous Cultures at Different Dilution Rates

  • Kim, Chang-Joon;Chang, Yong-Keun;Chun, Gie-Taek;Jeong, Yeon-Ho;Lee, Sang-Jong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.557-562
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    • 2002
  • Physiological characteristics such as specific productivity, morphology of Streptomyces cells Immobilized on celite beads, and operational stability at different dilution rates were investigated in continuous immobilized-cell cultures for the production of kasugamycin. At a dilution rate (D) of 0.05 $h^{-1}$, a relatively high specific productivity was attained and the loss of cell-loaded beads was negligible. At D=0.1 $h^{-1}$, a higher specific productivity and cell concentration could be obtained, resulting in a significantly improved volumetric kasugamycin productivity. However, no stable operation could be maintained due to a significant loss of cell-loaded beads from the reactor that was caused by their fluffy morphology developed in the later stage. At D=0.2 $h^{-1}$, the production of kasugamycin and cell growth were observed to be severely inhibited by the high concentration of residual maltose.

Global analysis of ginsenoside Rg1 protective effects in β-amyloid-treated neuronal cells

  • Shim, Ji Seon;Song, Min-Young;Yim, Sung-Vin;Lee, Seung-Eun;Park, Kang-Sik
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.566-571
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    • 2017
  • Background: A number of reports have described the protective effects of ginsenoside Rg1 (Rg1) in Alzheimer's disease (AD). However, the protective mechanisms of Rg1 in AD remain elusive. Methods: To investigate the potential mechanisms of Rg1 in ${\beta}$-amyloid peptide-treated SH-SY5Y cells, a comparative proteomic analysis was performed using stable isotope labeling with amino acids in cell culture combined with nano-LC-MS/MS. Results: We identified a total of 1,149 proteins in three independent experiments. Forty-nine proteins were significantly altered by Rg1 after exposure of the cells to ${\beta}$-amyloid peptides. The protein interaction network analysis showed that these altered proteins were clustered in ribosomal proteins, mitochondria, the actin cytoskeleton, and splicing proteins. Among these proteins, mitochondrial proteins containing HSD17B10, AARS2, TOMM40, VDAC1, COX5A, and NDUFA4 were associated with mitochondrial dysfunction in the pathogenesis of AD. Conclusion: Our results suggest that mitochondrial proteins may be related to the protective mechanisms of Rg1 in AD.

Identification and Characterization of Phytochrome-Regulated Phospholipase D in Oat Cells (Avena sativa L.)

  • Park, Cheon;Park, Moon-Hwan;Chae, Quae
    • BMB Reports
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    • v.29 no.6
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    • pp.535-539
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    • 1996
  • The activation of phospholipase D (PLD) catalyzes hydrolysis of phosphatidylcholine (PC) to phosphatidic acid (PA) and choline in plants as well as animals. To determine the presence of PLD in oat cells, we prepared inside-out plasma membrane and cytosolic fractions from oat tissues. PLD activities in both cytosol and plasma membrane were detected by ion chromatography method. The activity of PLD in plasma membrane was dependent upon $Ca^{2+}$ concentration and was heat stable. To investigate whether G-protein couples to PLD, the effects of $GTP{\gamma}S$ and $GDP{\beta}S$ on the PLD activity were measured. PLD activity was dramatically increased 300~400% in the presence of 50 ${\mu}M$ $GTP{\gamma}S$ but not in the presence of 50 ${\mu}M$ $GDP{\beta}S$. These results indicate that G-protein may be involved in regulation of PLD activity. To identify whether PLD is regulated by red light receptor, phytochrome, we irradiated red, far-red, or red/far-red/red light on oat protoplasts. PLD activity has increased 5-fold and 3-fold by treatment with red light and red/far-red/red light, respectively. In contrast, irradiation with far-red light had little or no effect on PLD activity. These results suggest that phytochrome regulates PLD activity through activation of G-protein in oat cells.

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Intrathoracic Desmoid Tumor Mimicking Pleural Mass: A Case Report

  • Kim, Na Rae;Chung, Dong-Hae;Lee, Jae-Ik;Jeong, Sung Hwan;Ha, Seung-Yeon
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.5
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    • pp.449-453
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    • 2009
  • Desmoid tumor (fibromatosis) is a histologically benign fibrous neoplasm showing locally infiltrating growth. This type of tumor commonly occurs in the abdomen, but intrathoracic desmoid tumor is uncommon. To date, 12 cases of intrathoracic desmoid tumor protruding into the pleural cavity, radiologically mimicking pleural masses, have been reported. Here, we report on a case of intrathoracic desmoid tumor protruding into the pleural cavity, and partially covered by parietal pleura. The main preoperative differential diagnoses included pleural solitary fibrous tumor, inflammatory pseudotumor or malignant mesothelioma. A near-total mass excision was performed. Pathologically, the tumor was composed of a paucicellular arrangement of spindle-shaped cells with fibromyxoid stroma. The resection margin was partially involved with spindle cells present. On histochemical staining, the spindle cells were strongly positive for vimentin and negative for CD34, consistent with a desmoid tumor. The patient was stable without further adjuvant treatment during 6-years of follow-up.

Repeated-batch Culture of Immobilized Gibberella fujikuroi B9 for Gibberellic Acid Production: An Optimization Study

  • Kim, Chang-Joon;Lee, Sang-Jong;Chang, Yong-Keun;Chun, Gie-Taek;Jeong, Yeon-Ho;Kim, Sung-Bae
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.6
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    • pp.544-549
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    • 2006
  • The performance of immobilized fungal cells on celite beads for the production of gibberrelic acid was investigated in flasks and 7-L stirred-tank reactor. Repeated incubations of immobilized fungal cells increased cell concentrations and volumetric productivity. The maximum volumetric productivity obtained in the immobilized-cell culture was 3-fold greater than that in suspended-cell culture. The concentration of cotton seed flour (CSF), among the various nutrients supplied, most significantly influenced productivity and operational stability. Notably, insoluble components in CSF were found to be essential for production. CSF at 6 g/L with 60 g/L glucose was found to be optimal for gibberellic acid production and stable operation by preventing excessive cell growth.

Hydrogen Sulfide Removal by Immobilized Thiobacillus novellas on $SiO_2$ in a Fluidized Bed Reactor

  • Cha, Jin-Myung;Shin, Hyun-Jae;Roh, Sung-Hee;Kim, Sun-Il
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.320-324
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    • 2007
  • The removal of hydrogen sulfide ($H_2S$) from aqueous media was investigated using Thiobacillus novellas cells immobilized on a $SiO_2$ carrier (biosand). The optimal growth conditions for the bacterial strain were $30^{\circ}C$ and initial pH of 7.0. The main product of hydrogen sulfide oxidation by T. novellus was identified as the sulfate ion. A removal efficiency of 98% was maintained in the three-phase fluidized-bed reactor, whereas the efficiency was reduced to 90% for the two-phase fluidized-bed reactor and 68% for the two-phase reactor without cells. The maximum gas removal capacity for the system was 254 g $H_2S/m^3/h$ when the inlet $H_2S$ loading was $300g/m^3/h(1,500ppm)$. Stable operation of the immobilized reactor was possible for 20 days with the inlet $H_2S$ concentration held to 1,100 ppm. The fluidized bed bioreactor appeared to be an effective means for controlling hydrogen sulfide emissions.