• Title/Summary/Keyword: Cell isolation

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Isolation and morphological characterization of natural killer cell in the sprague-dawley(SD) rats (Sprague-dawley(SD) 랫드에서 natural killer cell의 분리 ·동정 및 형태적 특징)

  • Kang, Kyung-sun;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.245-250
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    • 1992
  • This study was performed to demonstrate the presence of large granular lymphocyte(LGL) in Sprague-Dawley(SD) rats and morphologically observe NK cell and also establish the method of isolation of natural killer cell in SD rats. By percoll discontinuous density gradients centrifugation, highly enriched LGL population were shown to fraction 2(border line between 44.2% and 50.8%). LGL were shown to bind selectively to YAC1 mouse lymphoma cell. This fraction expressed very high NK cell cytolysis. Therefore, we thought that LGL have NK activity in SD rats. The Morphology of rat LGL is very similar to that of human LGL. These cells have an eccentric kidney-shaped nucleus. Their most distinctive feature was their cytoplasmic azurophilic granules. Another distinguishing feature of rat LGL was their high cytoplasmic : nuclear ratio. It was concluded that LGL played a role part in mediating natural killer activity in this species.

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Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

  • Park, Kyunghyuk;Frost, Jennifer M.;Adair, Adam James;Kim, Dong Min;Yun, Hyein;Brooks, Janie S.;Fischer, Robert L.;Choi, Yeonhee
    • Molecules and Cells
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    • v.39 no.10
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    • pp.768-775
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    • 2016
  • The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

High Efficiency High-Step-up Single-ended DC-DC Converter with Small Output Voltage Ripple

  • Kim, Do-Hyun;Kim, Hyun-Woo;Park, Joung-Hu;Jeon, Hee-Jong
    • Journal of Power Electronics
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    • v.15 no.6
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    • pp.1468-1479
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    • 2015
  • Renewable energy resources such as wind and photovoltaic power generation systems demand a high step-up DC-DC converters to convert the low voltage to commercial grid voltage. However, the high step-up converter using a transformer has limitations of high voltage stresses of switches and diodes when the transformer winding ratio increases. Accordingly, conventional studies have been applied to series-connect multioutput converters such as forward-flyback and switched-capacitor flyback to reduce the transformer winding ratio. This paper proposes new single-ended converter topologies of an isolation type and a non-isolation type to improve power efficiency, cost-effectiveness, and output ripple. The first proposal is an isolation-type charge-pump switched-capacitor flyback converter that includes an extreme-ratio isolation switched-capacitor cell with a chargepump circuit. It reduces the transformer winding number and the output ripple, and further improves power efficiency without any cost increase. The next proposal is a non-isolation charge-pump switched-capacitor-flyback tapped-inductor boost converter, which adds a charge-pump-connected flyback circuit to the conventional switched-capacitor boost converter to improve the power efficiency and to reduce the efficiency degradation from the input variation. In this paper, the operation principle of the proposed scheme is presented with the experimental results of the 100 W DC-DC converter for verification.

Isolation and Biological Activity of Verbascoside, A Potent Inhibitor of Protein Kinase C from the Calyx of Campsis grandiflora (능소화의 꽃받침으로부터 Protein Kinase C 저해물질인 Verbascoside의 분리 및 그 생물활성)

  • 이현선;박문수;오원근;안순철;김보연;김환묵;오구택;민태익;안종석
    • YAKHAK HOEJI
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    • v.37 no.6
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    • pp.598-604
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    • 1993
  • The calyx extract of Campsis grandiflora displayed inhibitory activity against protein kinase C from the bovine brain. Separation guided by protein kinase C enzyme assay and bleb forming assay led to isolation of a potent protein kinase C inhibitor that was identified as a known phenylpropanoid glycoside, verbascoside. It suppressed completely bleb-formation of K562 cell surface induced by phorbol 12,13-dibutylate at the concentration of 60 $\mu\textrm{g}$/ml and IC$_{50}$ of the protein kinase C occured at 20 $\mu{M}$. This compound was tested for cytotoxic activity against ten human tumor cell lines in vitro. it exhibited moderate cytotoxic activity against skin tumor cell line M14 (IC$_{50}$ 2.2 $\mu\textrm{g}$/ml) and very weak cytotoxicity against other cell lines (IC$_{50}$>10 $\mu\textrm{g}$/ml)

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Isolation of bovine Coronavirus from calves with diarrhea (소 Coronavirus 분리에 관한 연구)

  • 안재문;유기조;이종인
    • Korean Journal of Veterinary Service
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    • v.20 no.2
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    • pp.195-203
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    • 1997
  • Fecal samples were collected from 47 calves with diarrhea and 12 clinically normal co-h-abitants, and tested for virus using MDBK cell cultures. Three cytopathic viruses were isolated from 8 fecal samples obtained from diarrheic calves. The isolated viruses were neutralized by bovine coronavirus hyperimmune serum In plaque reduction assay and were detected in the cytoplasm of MDBK cell by bovine coronavirus hyperimmune serum using immunofluorescence staining. The viruses agglutinated mouse erythrocytes only among the various animal erythrocytes tested and new isolates were identified as bovine coronavirus.

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HOT CELL RENOVATION IN THE SPENT FUEL CONDITIONING PROCESS FACILITY AT THE KOREA ATOMIC ENERGY RESEARCH INSTITUTE

  • YU, SEUNG NAM;LEE, JONG KWANG;PARK, BYUNG SUK;CHO, ILJE;KIM, KIHO
    • Nuclear Engineering and Technology
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    • v.47 no.6
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    • pp.776-790
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    • 2015
  • Background: The advanced spent fuel conditioning process facility (ACPF) of the irradiated materials examination facility (IMEF) at the Korea Atomic Energy Research Institute (KAERI) has been renovated to implement a lab scale electrolytic reduction process for pyroprocessing. The interior and exterior structures of the ACPF hot cell have been modified under the current renovation project for the experimentation of the electrolytic reduction process using spent nuclear fuel. The most important aspect of this renovation was the installation of the argon compartment within the hot cell. Method: For the design and system implementation of the argon compartment system, a full-scale mock-up test and a three-dimensional (3D) simulation test were conducted in advance. The remodeling and repairing of the process cell (M8a), the maintenance cell (M8b), the isolation room, and their utilities were also planned through this simulation to accommodate the designed argon compartment system. Results and conclusion: Based on the considered refurbishment workflow, previous equipment in the M8 cell, including vessels and pipes, were removed and disposed of successfully after a zoning smear survey and decontamination, and new equipment with advanced functions and specifications were installed in the hot cell. Finally, the operating area and isolation room were also refurbished to meet the requirements of the improved hot cell facility.

Production of Trench Epitaxial Transistor(TETC) (Trench Epitaxial Transistor Cell(TETC)의 제조)

  • Yi, Cheon-Hee
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.26 no.8
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    • pp.1290-1298
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    • 1989
  • A new dynamic RAM cell called Trench Epitaxial Transistor Cell (TETC) has been developed for 4M to 16M DRAMS. Also the fabrication process for device isolation which can decrease the narrow effect using SEG process has been developed. We verified the characteristic of the new cell structure with the PICSES simulator on VAX8450.

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Study on the Evaluation Method of Electrical Isolation Property for Hydrogen Fuel Cell Vehicle (수소연료전지자동차의 절연성능 평가방법에 관한 고찰)

  • Lee, Ki-Yeon;Kim, Dong-Ook;Kim, Hyang-Kon
    • Transactions of the Korean hydrogen and new energy society
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    • v.22 no.6
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    • pp.810-818
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    • 2011
  • This paper analyzed the suitability about the isolation performance criteria which was based on human impedance and effect of current in IEC 60479-1 on the safety of human being was examined. The method of evaluation by megger and DC voltmeter was analyzed. The differences of isolation performance according to design of high-voltage system were analyzed. The factors which affect the insulation performance were analyzed for HFCV, EV, HEV, etc. through analysis of the isolation performance evaluation method. Finally, design for improved isolation performance was proposed.

Isolation, Culture and Identification of Porcine Skeletal Muscle Satellite Cells

  • Li, Bo-jiang;Li, Ping-hua;Huang, Rui-hua;Sun, Wen-xing;Wang, Han;Li, Qi-fa;Chen, Jie;Wu, Wang-jun;Liu, Hong-lin
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.8
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    • pp.1171-1177
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    • 2015
  • The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.

Identification of a Technique Optimized for the Isolation of Spermatogonial Stem Cells from Mouse Testes

  • Han, Na Rae;Park, Hye Jin;Lee, Hyun;Yun, Jung Im;Choi, Kimyung;Lee, Eunsong;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.327-336
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    • 2018
  • To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.