• Title/Summary/Keyword: Two-cell

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Construction of Glomerular Epithelial Cells Expressing Both Immune Tolerance and GFP Genes and Application to Cell Therapy by Cell Transplantation

  • Ohga, Masahiro;Ogura, Mariko;Matsumura, Mastoshi;Wang, Pi-Chao
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.5
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    • pp.303-310
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    • 2002
  • Cell therapy applied to wound healing or tissue regeneration presents a revolutionary realm to which principles of gene engineering and delivery may be applied. One promising application is the transplantation of cells into the wounded tissue to help the tissue repair. However, when cells are transplanted from in vitro to in vivo, immune rejection occurs due to the immune response triggered by the activation of T-cell, and the transplanted cells are destroyed by the attack of activated T-cell and lose their function. Immune suppressant such as FK506 is commonly used to suppress immune rejection during transplantation. However, such kind of immune suppressants not only suppresses immune rejection in the periphery of transplanted cells but also suppresses whole immune response system against pathogenic infection. In order to solve this problem, we developed a method to protect the desired cells from immune rejection without impairing whole immune system during cell transplantation. Previously, we reported the success of constructing glomerular epithelial cells for removal of immune complex, in which complement receptor of type 1 (CR1) was over-expressed on the membrane of renal glomerular epithelial cells and could bind immune complex of DNA/anti-DNA-antibody to remove immune complex through phagocy-tosis [1]. Attempting to apply the CR1-expressing cells to cell therapy and evade immune rejection during cell transplantation, we constructed three plasmids containing genes encoding a soluble fusion protein of cytolytic T lymphocyte associated antigen-4 (CTLA4Ig) and an enhanced green fluorescent protein (EGFP). The plasmids were transfected to the above-mentioned glomerular epithelial cells to express both genes simultaneously. Using the clone cells for cell transplantation showed that mice with autoimmune disease prolonged their life significantly as compared with the control mice, and two injections of the cells at the beginning of two weeks resulted in remarkable survivability, whereas it requires half a year and 50 administrations of proteins purified from the same amount of cells to achieve the same effect.

Mechanism of Differential Ag-specific Immune Induction by Different Tumor Cell Lysate Pulsed DC (종양 세포 용해액에 따른 수지상세포 유도 항원 특이 면역반응 차이의 기전 연구)

  • Lee, Kang-Eun;Shon, Hye-Jin;Kim, Myung-Joo;Baek, So-Young;Lee, Hyun-Ah
    • IMMUNE NETWORK
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    • v.6 no.3
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    • pp.145-153
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    • 2006
  • Background: Tumor cell lysate has been considered as a preferential antigen source for the therapeutic dendritic cell pulsing. Our experiences with in vivo study with animal tumor model indicate the tumor cell lysate dependent differential effect of DC therapy. Our previous data show that MC38 lysate pulsed-DC induced stronger ag-specific immunity than CT26 lysate pulsed-DC in vitro. In this study we tried to reveal the mechanism for differential induction of ag-specific immunity of different colon cancer cell lysate pulsed-DCs. Methods: MC38 and CT26 cell lines were prepared as lysate by freezing-thawing procedure. Tumor cell antigenicity was confirmed by detecting the surface expression of MHC I/II & B7.1/2 molecules. IL-10, IL-12 and TGF-beta in the tumor cell lysate were detected by ELISA and the presence of heat shock proteins were analysed by western blotting. Results: The secretion of IL-10, a immune-inhibitory cytokine was about 470% higher in CT26 lysate than in MC38. Hsp 70 was detected only in the MC38 lysate but not in the CT26. On the other hand, Hsp 60 and 90 expression were not different in two colon cancer cell lysates. Conclusion: In two different colon cancer cell lysate, immune inhibitory IL-10 (higher in CT26) and Hsp70 (MC38 superiority) were differentially expressed. These data indicate that higher agspecific immunity induction by MC38 lysate pulsed-DC may due to the expression of hsp70 and lower secretion of IL-10, a immune-inhibitory cytokine than CT26 lysate. The significance of other cytokine and the surface marker expression will be discussed.

Botryococcus braunii 배양에서 탄화수소의 two-stage 동시추출공정

  • An, Jin-Yeong;Choe, Jeong-Gyu;Sim, Sang-Jun;Kim, Byeong-U
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.229-232
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    • 2000
  • The carbon dioxide, nitrogen, and phosphate removals from wastewater using microalgae have extensively been studied. Especially, the green colonial algae Botryococcus braunii is characterized by unusual high hydrocarbon contents, ranging from 15 to 75% of dry weight, as long-chain unsaturated hydrocarbons. These hydrocarbons suggest that the possibility of renewable biofuels to be converted into useful fuels such as gasoline by simple catalytic cracking. The poor recovery (18 - 32%) of hydrocarbon from B. braunii culture in two-phase bubble column seems to be caused by insufficient mixing between two phases, which was operated using only aeration on the narrow interface between hydrophobic solvent and cell suspension. In addition, hydrocarbon was entrapped tightly in cell-matrix (formed by exopolysaccharide) of algal colony, which make difficult to extract using two-phase system. In order to overcome low recovery efficiency, two-stage extraction culture system including culture vessel and two-phase separator is now under development, resulted improving contact between solvent phase and cell suspension. Hydrocarbon recovery using this process was more than two times as that using two-phase extraction culture.

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A Modularized Two-Stage Charge Equalization Converter for Series Connected Lithium-Ion Battery Strings

  • Kim, Chol-Ho;Park, Hong-Sun;Moon, Gun-Woo
    • Proceedings of the KIPE Conference
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    • 2008.06a
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    • pp.535-537
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    • 2008
  • This paper proposes a modularized two-stage charge equalization converter for a series-connected lithium-ion battery string. In this paper, the series-connected battery sting is modularized into M modules, and each module has K cells in series. With this modularization, low voltage stress on the electronic devices can be achieved. A two-stage dc-dc converter with cell selection switches is employed. The first stage dc-dc converter steps down the high bus voltage to about 10 V. The second stage dc-dc converter integrated with selection switches equalizes the cell voltages. A prototype for 88 lithium-ion battery cells is optimally designed and implemented. Experimental results verify that the proposed equalization method has good cell balancing performance showing low voltage stress, small size, and low cost.

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Animal Cell Culture and the Production of Monoclonal Antibody(MAb) Using Biopolymer Membrane (생물고분자 막 형성을 이용한 동물세포 배양 및 단클론항체 생산)

  • 손정화;유선희;김성구
    • KSBB Journal
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    • v.13 no.1
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    • pp.13-19
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    • 1998
  • Biopolymer membrane was prepared using two oppositely charged natural biopolymers. The biopolymer membrane was used for the encapsulation of two hybridoma cell lines(ATCC CRL-1606, ATCC HB-8852) to produce monoclonal antibodies. In order to reduce the down stream steps, the pre size of the membrane was controlled to retain the monoclonal antibodies in the capsules based on the diffusion experiments with standard proteins. T-flask culture showed cell densities of 8$\times$107 cells/mL and 3$\times$107 cells/mL, and MAb concentrations of 506$\mu$g/mL and 109$\mu$g/mL for encapsulated ATCC CRL-1606 and HB-8852, respectively. Two liter perfusion cultures with encapsulated ATCC HB-8852 were performed to enhance the MAb production. The MAb production of the encapsulated hybridoma increased considerably comparing to the culture using silicon tubing for oxygen transfer.

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A High speed Standard Basis GF(2$^{m}$ ) Multiplier with A Known Primitive Coefficient Set (Standard Basis를 기반으로 하는 유한체내 고속 GF($2^m$) 곱셈기 설계)

  • 최성수;이영규;박민경;김기선
    • Proceedings of the IEEK Conference
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    • 1999.06a
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    • pp.333-336
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    • 1999
  • In this paper, a new high speed parallel input and parallel output GF(2$^{m}$ ) multiplier based on standard basis is proposed. The concept of the multiplication in standard basis coordinates gives an easier VLSI implementation than that of the dual basis. This proposed algorithm and method of implementation of the GF(2$^{m}$ ) multiplication are represented by two kinds of basic cells (which are the generalized and fixed basic cell), and the minimum critical path with pipelined operation. In the case of the generalized basic cell, the proposed multiplier is composed of $m^2$ basic cells where each cell has 2 two input AND gates, 2 two input XOR gates, and 2 one bit latches Specifically, we show that the proposed multiplier has smaller complexity than those proposed in 〔5〕.

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Fabrication of a Dual-Gap Substrate Using the Replica-molding Technique for Transflective Liquid Crystal Displays

  • Kim, Yeun-Tae;Hong, Jong-Ho;Cho, Seong-Min;Lee, Sin-Doo
    • Journal of Information Display
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    • v.10 no.2
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    • pp.68-71
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    • 2009
  • A replica-molding method of fabricating a dual-gap substrate for transflective liquid crystal (LC) displays is demonstrated. The dual-gap substrate provides homeotropic alignment for the LC molecules without any surface treatment and embedded bilevel microstructure on one of the two surfaces to maintain different cell gaps between the transmissive and reflective subpixels. The proposed transflective LC cell shows no electro-optic disparity between two subpixels and reduces the panel thickness and weight by 30% compared to the conventional transflective LC cell, which has two glass substrates.

Encapsulated Animal Cell Culture for the Production of Monoclonal Antibody(MAb)

  • Kim, Sung-Koo;Son, Jeong-Hwan;Yu, Sun-Hee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.2 no.2
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    • pp.73-76
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    • 1997
  • Biopolymer membrane was prepared using two oppositely charged natural biopolymer. The biopolymer membrane was used for the encapsulation of two hybridoma cell lines(ATCC CRL-1606, ATCC BH-8852) to produce monoclonal antibodies. In order to reduce the down stream steps, the pore size of the membrane was controlled to retain the monoclonal antibodies in the capsules based on the diffusion experiments with standard proteins. T-flask culture showed cell densities of 8$\times$107cells/mL 3$\times$107cells/mL, and MAb concentrations of 506 $\mu\textrm{g}$/mL and 109$\mu\textrm{g}$/mL for encapsulated ATCC CRL-1606 and HB-8852, respectively. Two liter perfusion culture with encapsulated ATCC HB-8852 was performed to enhance the MAb production. The MAb production of the encapsulated hybridoma increased considerably comparing to the culture using silicone tubing for oxygen transfer.

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Plant Cells on Earth and in Space

  • Braun, Markus;Sievers, Andreas
    • Animal cells and systems
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    • v.4 no.3
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    • pp.201-214
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    • 2000
  • Two quite different types of plant cells are analysed with regard to transduction of the gravity stimulus: (i) Unicellular rhizoids and protonemata of characean green algae; these are tube-like, tip-growing cells which respond to the direction of gravity. (ii) Columella cells located in the center of the root cap of higher plants; these cells (statocytes) perceive gravity. The two cell types contain heavy particles or organelles (sataoliths) which sediment in the field of gravity, thereby inducing the graviresponse. Both cell types were studied under microgravity conditions ($10^{-4}$/ g) in sounding rockets or spacelabs. From video microscopy of living Chara cells and different experiments with both cell types it was concluded that the position of statoliths depends on the balance of two forces, i.e. the gravitational force and the counteracting force mediated by actin microfilaments. The actomyosin system may be the missing link between the gravity-dependent movement of statoliths and the gravity receptor(s); it may also function as an amplifier.

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Two-Cell Spheroid Angiogenesis Assay System Using Both Endothelial Colony Forming Cells and Mesenchymal Stem Cells

  • Shah, Sajita;Kang, Kyu-Tae
    • Biomolecules & Therapeutics
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    • v.26 no.5
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    • pp.474-480
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    • 2018
  • Most angiogenesis assays are performed using endothelial cells. However, blood vessels are composed of two cell types: endothelial cells and pericytes. Thus, co-culture of two vascular cells should be employed to evaluate angiogenic properties. Here, we developed an in vitro 3-dimensional angiogenesis assay system using spheroids formed by two human vascular precursors: endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs). ECFCs, MSCs, or ECFCs+MSCs were cultured to form spheroids. Sprout formation from each spheroid was observed for 24 h by real-time cell recorder. Sprout number and length were higher in ECFC+MSC spheroids than ECFC-only spheroids. No sprouts were observed in MSC-only spheroids. Sprout formation by ECFC spheroids was increased by treatment with vascular endothelial growth factor (VEGF) or combination of VEGF and fibroblast growth factor-2 (FGF-2). Interestingly, there was no further increase in sprout formation by ECFC+MSC spheroids in response to VEGF or VEGF+FGF-2, suggesting that MSCs stimulate sprout formation by ECFCs. Immuno-fluorescent labeling technique revealed that MSCs surrounded ECFC-mediated sprout structures. We tested vatalanib, VEGF inhibitor, using ECFC and ECFC+MSC spheroids. Vatalanib significantly inhibited sprout formation in both spheroids. Of note, the $IC_{50}$ of vatalanib in ECFC+MSC spheroids at 24 h was $4.0{\pm}0.40{\mu}M$, which are more correlated with the data of previous animal studies when compared with ECFC spheroids ($0.2{\pm}0.03{\mu}M$). These results suggest that ECFC+MSC spheroids generate physiologically relevant sprout structures composed of two types of vascular cells, and will be an effective pre-clinical in vitro assay model to evaluate pro- or anti-angiogenic property.