• Title/Summary/Keyword: Adherent cells

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Hepatocyte Growth Factor is the Key Cytokine in Stimulating Potential Stem Cells in the Cord Blood into Hepatic Lineage Cells

  • Ryu, Kyung-Ha;Cho, Su-Jin;Woo, So-Youn;Seoh, Ju-Young;Jung, Yun-Jae;Han, Ho-Seong
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.117-123
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    • 2007
  • Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

Effect of ZNimesulide on the Differentiation and Survival of Endothelial Progenitor Cells

  • Oh, Ho-Kyun;Kim, Sun-Yong;Baek, Sang-Hong;Lim, Sung-Cil;Ahn, Hyun-Young;Shin, Jong-Chul;Hong, Sung-Hee;Hong, Yong-Kil;Joe, Young-Ae
    • Biomolecules & Therapeutics
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    • v.12 no.4
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    • pp.221-227
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    • 2004
  • Nonsteroidal anti-inflammatory drugs (NSAIDs), particularly the highly selective cyclooxygenase (COX)-2 inhibitors have been shown to decrease the growth of tumor, in part, by inhibition of neovascularization. Recently, besides mature endothelial cells, endothelial progenitor cells (EPCs) have been shown to contribute neovascularization in angiogenic tissues. In this study, we addressed a question whether nimesulide, a selective COX-2 inhibitor, could affect differentiation of EPCs into adhesive endothelial cells in vitro. Total mononuclear cells were isolated from cord blood by Ficoll density gradient centrifugation, and then the cells were incubated with nimesulide or vehicle control for 7 days. The number of adherent and spindle-shaped cells decreased by nimesulide treatment in a concentration-dependent fashion at a concentration range of 5 - 200 ${\mu}M$. Moreover, the adherent cells double positive for DiI-ac-LDL uptake and lectin binding significantly decreased upon nimesulide treatment. There was no change of expression of CD31 between treatment and control groups, whereas slight reduction was detected upon treatment in expression of VE-cadherin, ICAM-1, vWF, ${\alpha}v$, and ${\alpha}5$. Nimesulide also reduced cell viability during first 3 days' culture and induced apoptosis in adherent EPCs, resulting in increased annexin-V-positive and propidium iodide-negative cells. Taken together, these results suggest that nimesulide could be applied for the inhibition of new vessel formation, in part, by inhibiting differentiation and survival of EPCs.

Adhesion-induced generation of oxygen free radical from human alveolar macrophages and its mechanisms (폐포대식세포의 부착에 의한 산소유리기 분비능 활성화 및 그 기전)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.2
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    • pp.210-220
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    • 1996
  • Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.

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Protein Profiles Associated with Anoikis Resistance of Metastatic MDA-MB-231 Breast Cancer Cells

  • Akekawatchai, Chareeporn;Roytrakul, Sittiruk;Kittisenachai, Suthathip;Isarankura-Na-Ayudhya, Patcharee;Jitrapakdee, Sarawut
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.581-590
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    • 2016
  • Resistance to anoikis, a cell-detachment induced apoptosis, is one of the malignant phenotypes which support tumor metastasis. Molecular mechanisms underlying the establishment of this phenotype require further investigation. This study aimed at exploring protein expression profiles associated with anoikis resistance of a metastatic breast cancer cell. Cell survival of suspension cultures of non-metastatic MCF-7 and metastatic MDA-MB-231 cells were compared with their adherent cultures. Trypan blue exclusion assays demonstrated a significantly higher percentage of viable cells in MDA-MB-231 than MCF-7 cell cultures, consistent with analysis of annexin V-7-AAD stained cells indicating that MDA-MB-231 possess anti-apoptotic ability 1.7 fold higher than MCF-7 cells. GeLC-MS/MS analysis of protein lysates of MDA-MB-231 and MCF-7 cells grown under both culture conditions identified 925 proteins which are differentially expressed, 54 of which were expressed only in suspended and adherent MDA-MB-231 but not in MCF-7 cells. These proteins have been implicated in various cellular processes, including DNA replication and repair, transcription, translation, protein modification, cytoskeleton, transport and cell signaling. Analysis based on the STITCH database predicted the interaction of phospholipases, PLC and PLD, and 14-3-3 beta/alpha, YWHAB, with the intrinsic and extrinsic apoptotic signaling network, suggesting putative roles in controlling anti-anoikis ability. MDA-MB-231 cells grown in the presence of inhibitors of phospholipase C, U73122, and phospholipase D, FIPI, demonstrated reduced ability to survive in suspension culture, indicating functional roles of PLC and PLD in the process of anti-anoikis. Our study identified intracellular mediators potentially associated with establishment of anoikis resistance of metastatic cells. These proteins require further clarification as prognostic and therapeutic targets for advanced breast cancer.

Inhibitory Effect of Lactobacillus plantarum K11 on the Adhesion of Escherichia coli O157 to Caco-2 Cells

  • Lim, Sung-Mee;Ahn, Dong-Hyun;Im, Dong-Soon
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.343-349
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    • 2009
  • Inhibitory effect of Escherichia coli O157 adhered to Caco-2 cells by the cells of Lactobacillus plantarum K11 and the cell-free culture supernatant (CFCS) and bacteriocin prepared from this strain was investigated. As the cell counts of viable L. plantarum K11 previously adhered to Caco-2 were increased, the rate of adhesion and adherent cell counts of E. coli O157 was lower. However, because the heated L. plantarum K11 rarely have the adhesion ability to Caco-2, the adhesion rate and adherent cell counts of E. coli O157 were high. In addition, the inhibitory effects of E. coli O157 adhesion by the CFCS and bacteriocin of L. plantarum K11 were dose-dependent manner. Therefore, the inhibition of adhesion of E. coli O157 to Caco-2 may result from the antimicrobial substances such as lactic acid and bacteriocin. Moreover the inhibitory activity of adhesion by the heated bacteriocin for 30 min at 100oC was similar to activity of non-treated bacteriocin, but the activity was disappeared by treatment with protease.

Studies on the Activities of Peritoneal Macrophages Induced by Yookmijihwangtang, Palmijihwangtang and Gamijihwangtang (육미지황탕(六味地黃湯), 팔미지황탕(八味地黃湯) 및 가미지황탕(加味地黃湯)이 생쥐의 복강대식세포(腹腔大食細胞) 활성(活性)에 미치는 영향(影響))

  • Hahn, Il-Soo;Kim, Cheol-Joong
    • Journal of Haehwa Medicine
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    • v.6 no.1
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    • pp.331-347
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    • 1997
  • In order to investigate the effects of YOOKMIJIHWANGTANG, GAMIJIHWANGTANG and PALMIJIHWANGTANG on the various immune responses, the chemotactic, adherent, nitric oxide release ability of macrophages were studied in vivo and in vitro. The results obtained were follows : 1. The total number of peritoneal exudate cells from mice injected with three drugs were 2~3 times more than those from the PBS. 2. The administration of three drugs enhanced significantly the Fc receptor mediated activity(phagocytosis, rosette formation activity). 3. The administration of three drugs enhanced significantly the chemotactic activity of peritoneal macrophages. 4. The administration of three drugs enhanced significantly the adherent activity of macrophages and neutrophils. 5. The administration of GAMIJIHWANGTANG and PALMIJIHWANGTANG enhanced significantly the chemotatic activity of macrophages (p<0.01). And also the administration of YOOKMIJIHWANGTANG enhanced significantly the chemotatic activity of macrophages(p<0.05). 6. The administration of GAMIJIHWANGTANG and enhanced significantly the chemotatic activity of neutrophils(p<0.01). And also the administration of YOOKMIJIHWANGTANG and PALMIJIHWANGTANG enhanced significantly the chemotatic activity of neutrophils(p<0.05). From above findings, it is suggested that YOOKMIJDTWANGTANG, GAMIJIHWANGTANG and PALMIJIHWANGTANG seems to produce the increase of immune response such as the Fc receptor activity, chemotactic activity, adherent activity, nitrate release of macrophages. From the above results, the GAMIJIHWANGTANG among three drugs may be the most useful drug having immunostimulating effects.

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Pulmonary paraganglioma: report of a case (폐의 부신경절종 1례 보고)

  • 김광호
    • Journal of Chest Surgery
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    • v.15 no.4
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    • pp.456-459
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    • 1982
  • A case of pulmonary paraganglioma in a 32-year-old woman is presented. The tumor was adherent to the anteromedial basal segmental branch of the left pulmonary artery and had round groups of monotonous tumor cells with faintly eosinophilic and granular cytoplasm. Also tumor cells were divided into lobules by fine fibrous septae. The patient made an uneventful recovery and no recurrence of the lesion has occurred during the subsequent one year.

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Preclinlcal Evaluation of Polysaccharides Extracted from Korean Red-ginseng as an Antineoplastic Immunostimulator (홍삼다당체의 항암면역증강작용 연구)

  • 김기환;정인성
    • Journal of Ginseng Research
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    • v.21 no.2
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    • pp.78-84
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    • 1997
  • As a part of our ongoing effort to develop new antineoplastic immunostimulator from natural sources, bioassay-directed fractionationn of polysaccharides from Korean red ginseng was carried out by observing the proliferation of marine spleen cells and the generation of lymphoklne activated killer (LAK) cells. The acidic polysaccharide fractions proliferated spleen cells and generated LAK cells in proportion to their acidity in vitro. The LAK cell which was induced by ginseng showed tumoricidal activity against both NK celt sensitive and insensitive tumor target cells without major histocompatibility (MHC) restriction. Adherent macrophages and CD4+helper T cells were involved in the generation of the LAK cells. The acidic polysaccharide from Korean red ginseng synerglzed with recombinant IL-2 (rIff-2) at lower than 3 U/ml. The optimal doses of the acidic polysaccharide from Korean red ginseng for the proliferation of spleen cells and for the generation of LAK cells were 1 mg/ml and 100 $\mu\textrm{g}$/ml, respectively; this means that the mechanisms for the both activities may be different from each other.

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Immunomodulatory activity of Salicornia herbacea L. Components

  • Im, Sun-A;Kim, Goo-Whan;Lee, Chong-Kil
    • Natural Product Sciences
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    • v.9 no.4
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    • pp.273-277
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    • 2003
  • Salicornia herbacea is an annual herb growing in salt marshes and on muddy seashores. Salicornia herbacea has been used as a fork medicine as well as a seasoned vegetable. In fork medicine, Salicornia herbacea has been used to treat a variety of diseases such as constipation, obesity, diabetes, asthma, arthritis and cancer. However, the biological mechanisms for these activities have not been characterized, nor the active components. The immunomodulatory activity of Salicornia herbacea components were studied in the present study. The components of Salicornia herbacea were prepared from the whole plant by passage through a fine screen, and then dialyzed against PBS overnight. Immunomodulatory activities of the Salicornia herbacea components were examined on a mouse macrophage cell line, RAW 264.7 cells. The Salicornia herbacea components were shown to stimulate cytokine production, nitric oxide release, and expression of surface molecules in a dose dependent manner. The Salicornia herbacea components also induced further differentiation of slightly adherent RAW 264.7 cell into strongly adherent macrophages. These results indicate that Salicornia herbacea contains immunomodulator(s) that induces activation of macrophages.

Comparison of alkaline phosphatase activity of MC3T3-E1 cells cultured on different Ti surfaces: modified sandblasted with large grit and acid-etched (MSLA), laser-treated, and laser and acid-treated Ti surfaces

  • Li, Lin-Jie;Kim, So-Nam;Cho, Sung-Am
    • The Journal of Advanced Prosthodontics
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    • v.8 no.3
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    • pp.235-240
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    • 2016
  • PURPOSE. In this study, the aim of this study was to evaluate the effect of implant surface treatment on cell differentiation of osteoblast cells. For this purpose, three surfaces were compared: (1) a modified SLA (MSLA: sand-blasted with large grit, acid-etched, and immersed in 0.9% NaCl), (2) a laser treatment (LT: laser treatment) titanium surface and (3) a laser and acid-treated (LAT: laser treatment, acid-etched) titanium surface. MATERIALS AND METHODS. The MSLA surfaces were considered as the control group, and LT and LAT surfaces as test groups. Alkaline phosphatase expression (ALP) was used to quantify osteoblastic differentiation of MC3T3-E1 cell. Surface roughness was evaluated by a contact profilometer (URFPAK-SV; Mitutoyo, Kawasaki, Japan) and characterized by two parameters: mean roughness (Ra) and maximum peak-to-valley height (Rt). RESULTS. Scanning electron microscope revealed that MSLA (control group) surface was not as rough as LT, LAT surface (test groups). Alkaline phosphatase expression, the measure of osteoblastic differentiation, and total ALP expression by surface-adherent cells were found to be highest at 21 days for all three surfaces tested (P<.05). Furthermore, ALP expression levels of MSLA and LAT surfaces were significantly higher than expression levels of LT surface-adherent cells at 7, 14, and 21 days, respectively (P<.05). However, ALP expression levels between MSLA and LAT surface were equal at 7, 14, and 21 days (P>.05). CONCLUSION. This study suggested that MSLA and LAT surfaces exhibited more favorable environment for osteoblast differentiation when compared with LT surface, the results that are important for implant surface modification studies.