• Title/Summary/Keyword: Cell surface properties

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Establishment and Characterization of Bone Marrow Mesenchymal Stromal/Stem Cells (MSCs) Derived from ${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) Pig (${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명)

  • Ock, Sun-A;Oh, Keon Bong;Hwang, Seongsoo;Im, Seoki;Kim, Youngim;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.281-287
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    • 2013
  • A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

$In$ $vitro$ MRI and Characterization of Rat Mesenchymal Stem Cells Transduced with Ferritin as MR Reporter Gene (페리틴 리포터 유전자를 발현하는 백서 중간엽 줄기세포의 특성과 자기공명영상 연구)

  • Shin, Cheong-Il;Lee, Whal;Woo, Ji-Su;Park, Eun-Ah;Kim, Pan-Ki;Song, Hyun-Bok;Kim, Hoe-Suk
    • Investigative Magnetic Resonance Imaging
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    • v.16 no.1
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    • pp.47-54
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    • 2012
  • Purpose : This study was performed to evaluate the characteristics of rat mesenchymal stem cells (RMSCs) transduced with human ferritin gene and investigate $in$ $vitro$ MRI detectability of ferritin-transduced RMSCs. Materials and Methods: The RMSCs expressing both myc-tagged human ferritin heavy chain subunit (myc-FTH) and green fluorescence protein (GFP) were transduced with lentiviurs. Transduced cells were sorted by GFP expression using a fluorescence-activated cell sorter. Myc-FTH and GFP expression in transduced cells were detected by immunofluorescence staining. The cell proliferative ability and viability were assessed by MTT assay. The RMSC surface markers (CD29+/CD45-) were analyzed by flow cytometry. The intracellular iron amount was measured spectrophotometically and the presence of ferritin-iron accumulation was detected by Prussian blue staining. $In$ $vitro$ magnetic resonance imaging (MRI) study of cell phantoms was done on 9.4 T MR scanner to evaluate the feasibility of imaging the ferritin-transduced RMSCs. Results: The myc-FTH and GFP genes were stably transduced into RMSCs. No significant differences were observed in terms of biologic properties in transduced RMSCs compared with non-transduced RMSCs. Ferritin-transduced RMSCs exhibited increased iron accumulation ability and showed significantly lower $T_2$ relaxation time than non-transduced RMSCs. Conclusion: Ferritin gene as MR reporter gene could be used for non-invasive tracking and visualization of therapeutic mesenchymal stem cells by MRI.

Change of Stratification of Three Dimensional Culture by Gingival Keratinocytes & Fibroblasts (치은 각화상피세포와 섬유아세포를 이용한 삼차원적 배양시 중층화 동안의 변화)

  • Jung, Tae-Heup;Hyun, Ha-Na;Kim, Yun-Sang;Kim, Eun-Cheol;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.129-142
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    • 2002
  • Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.

Characterization of Human Dental Pulp Cells from Supernumerary Teeth by Using Flow Cytometry Analysis (유세포 분석을 통한 과잉치 치수 유래 세포의 줄기세포 특성 연구)

  • You, Yonsook;Kim, Jongbin;Shin, Jisun;Lee, June-Haeng;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.46 no.3
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    • pp.337-342
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    • 2019
  • The aim of this study was to analyze cells from human dental pulp tissue of impacted supernumerary teeth as stem cells with flow cytometry. Human dental pulp cells from 15 supernumerary teeth were identified their characteristics as stem cells by expression of mesenchymal stem cell markers through flow cytometry analysis at passage 3 and passage 10. Cluster of differentiation (CD) 73, CD 90, CD 34, CD 45 and STRO-1 cell surface markers were used to figure out characteristics of dental pulp stem cells from supernumerary teeth. At passage 3, the cell population showed positive expression of CD 73, CD90 and STRO-1, lacked expression of CD 34 and CD 45. At passage 10, CD 73, CD 90 and STRO-1 showed positive expression while CD 34 and CD 45 showed negative expression. This study indicated that dental pulp stem cells of supernumerary teeth had the properties of mesenchymal stem cells at both early and late passage. Impacted supernumerary teeth could be considered as a noble source of stem cells because of rapid growth and maintaining characteristics of stem cells until late passage.

Effect of Cleaning Processes of Silicon Wafer on Surface Passivation and a-Si:H/c-Si Hetero-Junction Solar Cell Performances (기판 세정특성에 따른 표면 패시배이션 및 a-Si:H/c-Si 이종접합 태양전지 특성변화 분석)

  • Song, Jun-Yong;Jeong, Dae-Young;Kim, Chan-Seok;Park, Sang-Hyun;Cho, Jun-Sik;Song, Jin-Soo;Wang, Jin-Suk;Lee, Jeong-Chul
    • Korean Journal of Materials Research
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    • v.20 no.4
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    • pp.210-216
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    • 2010
  • This paper investigates the dependence of a-Si:H/c-Si passivation and heterojunction solar cell performances on various cleaning processes of silicon wafers. It is observed that the passivation quality of a-Si:H thin-films on c-Si wafers depends highly on the initial H-termination properties of the wafer surface. The effective minority carrier lifetime (MCLT) of highly H-terminated wafer is beneficial for obtaining high quality passivation of a-Si:H/c-Si. The wafers passivated by p(n)-doped a-Si:H layers have low MCLT regardless of the initial H-termination quality. On the other hand, the MCLT of wafers incorporating intrinsic (i) a-Si:H as a passivation layer shows sensitive variation with initial cleaning and H-termination schemes. By applying the improved cleaning processes, we can obtain an MCLT of $100{\mu}sec$ after H-termination and above $600{\mu}sec$ after i a-Si:H thin film deposition. By adapting improved cleaning processes and by improving passivation and doped layers, we can fabricate a-Si:H/c-Si heterojunction solar cells with an active area conversion efficiency of 18.42%, which cells have an open circuit voltage of 0.670V, short circuit current of $37.31\;mA/cm^2$ and fill factor of 0.7374. These cells show more than 20% pseudo efficiency measured by Suns-$V_{oc}$ with an elimination of series resistance.

Surface-Displayed Porcine IFN-λ3 in Lactobacillus plantarum Inhibits Porcine Enteric Coronavirus Infection of Porcine Intestinal Epithelial Cells

  • Liu, Yong-Shi;Liu, Qiong;Jiang, Yan-Long;Yang, Wen-Tao;Huang, Hai-Bin;Shi, Chun-Wei;Yang, Gui-Lian;Wang, Chun-Feng
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.515-525
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    • 2020
  • Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN-λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.

Performance of Carbon Cathode and Anode Electrodes Functionalized by N and O Doping Treatments for Charge-discharge of Vanadium Redox Flow Battery (탄소전극의 질소 및 산소 도핑에 따른 바나듐 레독스-흐름전지 양극 및 음극에서의 촉매화학적 특성 연구)

  • Lim, Hyebin;Kim, Jiyeon;Yi, Jung S.;Lee, Doohwan
    • Clean Technology
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    • v.23 no.3
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    • pp.308-313
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    • 2017
  • In this study, we investigated the electrocatalytic effects of the N and O co-doping of Graphite Felt (GF) electrode for the vanadium redox flow battery (VRFB) at the cathode and the anode reaction, respectively. The electrodes were prepared by chemical vapor deposition (CVD) with $NH_3-O_2$ at 773 K, and its effects were compared with an electrode prepared by an O doping treatment. The surface morphology and chemical composition of the electrodes were characterized by scanning electron microscopy (SEM) and photoelectron spectroscopy (XPS). The electrocatalytic properties of these electrodes were characterized in a VRFB single cell comparing the efficiencies and performance of the electrodes at the cathode, anode, and single cell level. The results exhibited about 2% higher voltage and energy efficiencies on the N-O-GF than the O-GF electrode. It was found that the N and O co-doping was particularly effective in the enhancement of the reduction-oxidation reaction at the anode.

A Study on the Vanadium Oxide Thin Films as Cathode for Lithium Ion Battery Deposited by RF Magnetron Sputtering (RF 마그네트론 스퍼터링으로 증착된 리튬 이온 이차전지 양극용 바나듐 옥사이드 박막에 관한 연구)

  • Jang, Ki-June;Kim, Ki-Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.6
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    • pp.80-85
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    • 2019
  • Vanadium dioxide is a well-known metal-insulator phase transition material. Lots of researches of vanadium redox flow batteries have been researched as large scale energy storage system. In this study, vanadium oxide($VO_x$) thin films were applied to cathode for lithium ion battery. The $VO_x$ thin films were deposited on Si substrate($SiO_2$ layer of 300 nm thickness was formed on Si wafer via thermal oxidation process), quartz substrate by RF magnetron sputter system for 60 minutes at $500^{\circ}C$ with different RF powers. The surface morphology of as-deposited $VO_x$ thin films was characterized by field-emission scanning electron microscopy. The crystallographic property was confirmed by Raman spectroscopy. The optical properties were characterized by UV-visible spectrophotometer. The coin cell lithium-ion battery of CR2032 was fabricated with cathode material of $VO_x$ thin films on Cu foil. Electrochemical property of the coin cell was investigated by electrochemical analyzer. As the results, as increased of RF power, grain size of as-deposited $VO_x$ thin films was increased. As-deposited thin films exhibit $VO_2$ phase with RF power of 200 W above. The transmittance of as-deposited $VO_x$ films exhibits different values for different crystalline phase. The cyclic performance of $VO_x$ films exhibits higher values for large surface area and mixed crystalline phase.

Preparation of superhydrophilic coating solutions containing fluorosurfactants and characterization of their antifogging and antifouling properties (불소계면활성제를 함유한 초친수 코팅액의 제조 및 방담 방오 특성)

  • Lee, Soo;Im, Sun Moon;Hwang, Heon
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.525-535
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    • 2017
  • In order to produce hydrophilic coating solution, which has superior antifogging and antifouling effect on the glass surface of solar cell module as well as improving photovoltaic efficiency, nanosilica was dispersed in an aqueous solution of Tween 20 and fluorosurfactant composed of decafluorobutane and polyethylene glycol. The antifogging effect at high temperature was excellent for all the coating solutions containing nanosilica, but the antifouling effect was observed when the content of nanosilica was over 6 wt%. As the content of fluorosurfactant increased, the initial water contact angle slightly increased and the antifogging effect remained well until 500 wiping with wet $Wipeol^{(R)}$. The antifouling effect was also excellent regardless of the content of fluorosurfactant, thus 0.1 wt% of the fluorosurfactant was enough for a coating solution production. From the AFM results, when 0.1 wt% to 0.3 wt% of the fluoro surfactant was added, the fractal structure of the coated glass surface was clearly existed and contributed to the better antifouling effect. The transmittance of coated glass surface was highest in TL-1 coating solution containing 0.1 wt% of fluorosurfactant, and the addition of fluorosurfactant in a larger amount than 0.1 wt% did not improve the transmittance. This result is in good agreement with the previous AFM result which shows a high surface roughness as well as a fractal structure formation for the TL-1 coating solution.

Study on the Improvement of Electrochemical Performance by Controlling the Surface Characteristics of the Oxygen Electrode Porous Transport Layer for Proton Exchange Membrane Water Electrolysis (양이온 교환막 수전해용 산화전극 확산층의 표면 특성 제어를 통한 전기화학적 성능 개선 연구)

  • Lee, Han Eol;Linh, Doan Tuan;Lee, Woo-kum;Kim, Taekeun
    • Applied Chemistry for Engineering
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    • v.32 no.3
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    • pp.332-339
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    • 2021
  • Recently, due to concerns about the depletion of fossil fuels and the emission of greenhouse gases, the importance of hydrogen energy technology, which is a clean energy source that does not emit greenhouse gases, is being emphasized. Water electrolysis technology is a green hydrogen technology that obtains hydrogen by electrolyzing water and is attracting attention as one of the ultimate clean future energy resources. In this study, the surface properties of the porous transport layer (PTL), one of the cell components of the proton exchange membrane water electrolysis (PEMWE), were controlled using a sandpaper to reduce overvoltage and increase performance and stability. The surfaces of PTL were sanded using sandpapers of 400, 180, and 100 grit, and then all samples were finally treated with the sandpaper of 1000 grit. The prepared PTL was analyzed for the degree of hydrophilicity by measuring the water contact angle, and the surface shape was observed through SEM analysis. In order to analyze the electrochemical characteristics, I-V performance curves and impedance measurements were conducted.