• 제목/요약/키워드: Adherent cells

검색결과 97건 처리시간 0.031초

Isolation of Mesenchymal Stem-like Cells from a Pituitary Adenoma Specimen

  • Shim, Jin-Kyoung;Kang, Seok-Gu;Lee, Ji-Hyun;Chang, Jong Hee;Hong, Yong-Kil
    • 대한의생명과학회지
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    • 제19권4호
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    • pp.295-302
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    • 2013
  • Some of the pituitary adenomas are invasive and spread into neighboring tissues. In previous studies, the invasion of pituitary adenomas is thought to be associated with epithelial-mesenchymal transition (EMT). In addition to that, we thought that mesenchymal stem cells (MSCs) exist in relevant microenvironment in pituitary adenoma. However, it has been little known about the existence of MSCs from pituitary adenoma. So we investigated whether mesenchymal stem-like cells (MSLCs) can be isolated from the pituitary adenoma specimen. We isolated and cultured candidate MSLCs from the fresh pituitary adenoma specimen with the same protocols used in culturing bone marrow derived MSCs (BM-MSCs). The cultured candidate MSLCs were analyzed by fluorescence-activated cell sorting (FACS) for surface markers associated with MSCs. Candidate MSLCs were exposed to mesenchymal differentiation conditions to determine the mesenchymal differentiation potential of these cells. To evaluate the tumorigenesis of candidate MSLCs from pituitary adenoma, we implanted these cells into the brain of athymic nude mice. We isolated cells resembling BM-MSCs named pituitary adenoma stroma mesenchymal stem-like cells (PAS-MSLCs). PAS-MSLCs were spindle shaped and had adherent characteristics. FACS analysis identified that the PAS-MSLCs had a bit similar surface markers to BM-MSCs. Isolated cells expressed surface antigen, positive for CD105, CD75, and negative for CD45, NG2, and CD90. We found that these cells were capable of differentiation into adipocytes, osteocytes and chondrocytes. Tumor was not developed in the nude mice brains that were implanted with the PAS-MSLCs. In this study, we showed that MSLCs can be isolated from a pituitary adenoma specimen which is not tumorigenic.

Toxoplasma 면역모체로 부터 출산된 신생마우스에 있어서 Vaccination 효능 저하 (Failure of Toxoplasma - Vaccination in Mice Born to Immune Mothers)

  • 이정호;하대유
    • 대한미생물학회지
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    • 제20권1호
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    • pp.103-107
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    • 1985
  • Female ICR mice were vaccinated against Toxoplasma gondii(RH strain), infected 2 weeks later and after recovery mated to normal ICR males. Control matings were with normal ICR females. The progeny of the above matings were weaned at 1 week, vaccinated at 1, 2, 3, 4 or 5 weeks of age and infected 2 weeks later with lethal Toxoplasma tachyzoites. As assessed by survival, the effectiveness of vaccination among offspring of vaccinated-recovered mothers was greatly impaired than that of control mice, with respective of age : where mice did survive, recovery was greatly delayed relative to the controls. The protective effect of vaccination among infants born to control mothers was also blocked by maternal specific antibodies, by administration of high-titered specific antibodies or by transfer of nylon wool adherent immune-spleen cells, but was augmented by transfer of nylon wool passed immune-spleen cells. These results indicate that this impairment of vaccination may be due to the transmission of maternal specific antibodies to the offspring which acts to suppress both priming by the vaccine and the generation of parasite-specific helper T cells.

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한우 림프구에 대한 단일항체 생산 1. T 림프구에 대한 단일항체 (Production of monoclonal antibody(MAb) for lymphocytes of Korean native cattle 1. MAb to T lymphocytes)

  • 송희종;운재호;김태중;채효석;허부홍;최민순
    • 대한수의학회지
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    • 제35권4호
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    • pp.769-776
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    • 1995
  • This study was undertaken to develop the monoclonal antibody(MAb) for lymphocytes of Korean native cattle by the cell hybridization of myeloma P3/NS-1/ 1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with nylon wool column eluted peripheral T lymphocytes of Korean native cattle. The isotype of MAb KCT-14 against T lymphocyte was mouse $IgG_1$. KCT-14 positivity of mononuclear cells(MNC) from peripheral blood lymphocytes, nylon wool nonadherent and adherent-lymphocytes was 41.7%, 58.4% and 22.6%, respectively. And that of mesenteric lymph node-, spleen and thymus-MNC was 43.3%, 40.2% and 33.6%, respectively. Immunoperoxidase staining of frozen tissue sections showed that the MAb positive cells were located in the medulla of the thymus and in the paracortical area and the mantle zone of the germinal center in the lymph nodes. These results indicated that KCT-14 was one of the MAb for investigate of T lymphocyte subpopulations in the Korean native cattle.

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Lysyl-tRNA Synthetase Inhibits Various Shear Stress-stimulated Signaling Pathways in Endothelial Cells

  • Park, Heon-Yong
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2008년도 Proceedings of the Convention
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    • pp.103-115
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    • 2008
  • Hemodynamic shear stress, the dragging force generated by blood flow, is known as an anti-atherogenic factor. We tested whether lysyl-tRNA synthetase (KRS) will be utilized as an agent controlling shear-sensing systems. KRS was previously known to be secreted as a pro-inflammatory agent. Here we found that KRS inhibited various shear-stimulated signaling pathways. We further found that KRS binds to detergent-resistant membrane (DRM), indicating that KRS binding molecules exist in DRM, specialized regions of the plasma membrane. DRM plays important roles in a variety of cellular processes and consists of gangliosides, signaling molecules and cytoskeletons. We then determined that KRS was colocalized with integrins ${\alpha}4$, ${\alpha}5$ and $av{\beta}3$. In addition, KRS was shown to be associated with sialic acid, existing at the end of gangliosides. Interestingly, the adherent effect of KRS was inhibited by pretreatment with sialic acid. Moreover, treatment of endothelial cells with neuraminidase appeared to inhibit both the KRS adhesion to endothelial cells and shear-stimulated signaling. In conclusion, KRS is likely to be utilized as a vascular regulator.

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Butyrate modulates bacterial adherence on LS174T human colorectal cells by stimulating mucin secretion and MAPK signaling pathway

  • Jung, Tae-Hwan;Park, Jeong Hyeon;Jeon, Woo-Min;Han, Kyoung-Sik
    • Nutrition Research and Practice
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    • 제9권4호
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    • pp.343-349
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    • 2015
  • BACKGROUND/OBJECTIVES: Fermentation of dietary fiber results in production of various short chain fatty acids in the colon. In particular, butyrate is reported to regulate the physical and functional integrity of the normal colonic mucosa by altering mucin gene expression or the number of goblet cells. The objective of this study was to investigate whether butyrate modulates mucin secretion in LS174T human colorectal cells, thereby influencing the adhesion of probiotics such as Lactobacillus and Bifidobacterium strains and subsequently inhibiting pathogenic bacteria such as E. coli. In addition, possible signaling pathways involved in mucin gene regulation induced by butyrate treatment were also investigated. MATERIALS/METHODS: Mucin protein content assay and periodic acid-Schiff (PAS) staining were performed in LS174T cells treated with butyrate at various concentrations. Effects of butyrate on the ability of probiotics to adhere to LS174T cells and their competition with E. coli strains were examined. Real time polymerase chain reaction for mucin gene expression and Taqman array 96-well fast plate-based pathway analysis were performed on butyrate-treated LS174T cells. RESULTS: Treatment with butyrate resulted in a dose-dependent increase in mucin protein contents in LS174T cells with peak effects at 6 or 9 mM, which was further confirmed by PAS staining. Increase in mucin protein contents resulted in elevated adherence of probiotics, which subsequently reduced the adherent ability of E. coli. Treatment with butyrate also increased transcriptional levels of MUC3, MUC4, and MUC12, which was accompanied by higher gene expressions of signaling kinases and transcription factors involved in mitogen-activated protein kinase (MAPK) signaling pathways. CONCLUSIONS: Based on our results, butyrate is an effective regulator of modulation of mucin protein production at the transcriptional and translational levels, resulting in changes in the adherence of gut microflora. Butyrate potentially stimulates the MAPK signaling pathway in intestinal cells, which is positively correlated with gut defense.

In Vitro Differentiation of Mesenchymal Progenitor Cells Derived from Porcine Umbilical Cord Blood

  • Kumar, Basavarajappa Mohana;Yoo, Jae-Gyu;Ock, Sun-A;Kim, Jung-Gon;Song, Hye-Jin;Kang, Eun-Ju;Cho, Seong-Keun;Lee, Sung-Lim;Cho, Jae-Hyeon;Balasubramanian, Sivasankaran;Rho, Gyu-Jin
    • Molecules and Cells
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    • 제24권3호
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    • pp.343-350
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    • 2007
  • Mesenchymal stem/progenitor cells (MPCs) were isolated from porcine umbilical cord blood (UCB) and their morphology, proliferation, cell cycle status, cell-surface antigen profile and expression of hematopoietic cytokines were characterized. Their capacity to differentiate in vitro into osteocytes, adipocytes and chondrocytes was also evaluated. Primary cultures of adherent porcine MPCs (pMPCs) exhibited a typical fibroblast-like morphology with significant renewal capacity and proliferative ability. Subsequent robust cell growth was indicated by the high percentage of quiescent (G0/G1) cells. The cells expressed the mesenchymal surface markers, CD29, CD49b and CD105, but not the hematopoietic markers, CD45 and CD133 and synthesized hematopoietic cytokines. Over 21 days of induction, the cells differentiated into osteocytes adipocytes and chondrocytes. The expression of lineage specific genes was gradually upregulated during osteogenesis, adipogenesis and chondrogenesis. We conclude that porcine umbilical cord blood contains a population of MPCs capable of self-renewal and of differentiating in vitro into three classical mesenchymal lineages.

Sun Ginseng Protects Endothelial Progenitor Cells From Senescence Associated Apoptosis

  • Im, Woo-Seok;Chung, Jin-Young;Bhan, Jae-Jun;Lim, Ji-Yeon;Lee, Soon-Tae;Chu, Kon;Kim, Man-Ho
    • Journal of Ginseng Research
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    • 제36권1호
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    • pp.78-85
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    • 2012
  • Endothelial progenitor cells (EPC) are a population of cells that circulate in the blood stream. They play a role in angiogenesis and, therefore, can be prognostic markers of vascular repair. Ginsenoside $Rg_3$ prevents endothelial cell apoptosis through the inhibition of the mitochondrial caspase pathway. It also affects estrogen activity, which reduces EPC senescence. Sun ginseng (SG), which is heat-processed ginseng, has a high content of ginsenosides. The purpose of this study was to investigate the protective effects of SG on senescence-associated apoptosis in EPCs. In order to isolate EPCs, mononuclear cells of human blood buffy coats were cultured and characterized by their uptake of acetylated low-density lipoprotein (acLDL) and their binding of Ulex europaeus agglutinin I (ulex-lectin). Flow cytometry with annexin-V staining was performed in order to assess early and late apoptosis. Senescence was determined by ${\beta}$-galactosidase (${\beta}$-gal) staining. Staining with 4'-6-Diamidino-2-phenylindole verified that most adherent cells (93${\pm}$2.7%) were acLDL-positive and ulex-lectin-positive. The percentage of ${\beta}$-gal-positive EPCs was decreased from 93.8${\pm}$2.0% to 62.5${\pm}$3.6% by SG treatment. A fluorescence-activated cell sorter (FACS) analysis showed that 4.9% of EPCs were late apoptotic in controls. Sun ginseng decreased the apoptotic cell population by 39% in the late stage of apoptosis from control baseline levels. In conclusion, these results show antisenescent and antiapoptotic effects of SG in human-derived EPCs, indicating that SG can enhance EPC-mediated repair mechanisms.

Proteolysis of $\beta$-Catenin in Apoptotic Jurkat Cells

  • Hwang, Sang-Gu;Park, Jeong-Uck;Lee, Hyung-Chul;Joo, Woo-Hong;Cho, Yong-Kweon;Moon, Ja-Young
    • Journal of Life Science
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    • 제10권1호
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    • pp.57-63
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    • 2000
  • ${\beta}$-catenin, which plays a critical role in both the cytoskeleton and in transcriptional regulation in variousadherent cell types, undergoes degradation during adherent cell apoptosis. Although ${\beta}$-catenin has been reported to be present in Jurkat T-acute lymphoblastic leukemia cells, the regulation of ${\beta}$-catenin in hematologic malignancies have not been examined. The data presented here demonstrate that treatment of the T cell leukemia Jurkat iwht the apoptosis inducer anti-Fas induced proteolytic cleavage of ${\beta}$-catenin. ${\beta}$-catenin was cleaved at both the N- and C-terminus after anti-Fas treatment. Cleavage of intact ${\beta}$-catenin was completely inhibited by caspase selective protease inhibitors. These data demonstrate that ${\beta}$ -catenin proteolysis is triggered by the cross-linking of the Fas receptor on Jurkat cells and subsequent activation of caspase protease. There was a clear accumulatio of the large proteolytic fragment in Jurkat cells treated with lactacystin of ALLM. These are potent inhibitors of proteasome and calpain. these results suggest that both the proteasome and clapain may recognize the large ${\beta}$-catenin fragment as a substrate fot further degradation and that these pathewasy may act downstream of scapase in response to Fas receptor activation. Therefore, we suggest that ${\beta}$-catenin may play a role in promoting Jurkat survival.

제브라피쉬(Danio rerio) 배아로부터 동형세포주 확립 (Establishment and Characterization of Clonal Cell Lines from Zebrafish, Danio rerio)

  • 이기영
    • 한국어류학회지
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    • 제20권1호
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    • pp.1-6
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    • 2008
  • 제브라피쉬 배아로부터 확보한 세포주로부터 외부형태와 세포 크기에 따라 3종류의 동형세포주를 확립하였다. 활발하게 증식하는 안정된 세포주 및 이들로부터 확립된 동형세포주의 세포특성은 변하지 않고 지속적으로 유지되었으며, 안정된 세포주로부터 총 18개의 콜로니를 확보하여 배양한 다음 3종류의 동형세포주를 선별하여 세포 특성을 분석하였다. 대부분의 동형세포주는 약 80% 정도 정상적인 염색체(2N=50)를 가지고 있었으며 FACs 분석과 일치하였다. 배아로부터 확립된 동형세포주에 항체테스트 결과, vimentin에서 양성을 보이는 결과로 볼 때 확립된 세포주는 분화된 섬유세포임이 확인되었다. 이러한 결과는, 확립된 동형세포주를 이용한 유전자조작과 어류복제에의 활용도를 높일 수 있음을 시사한다.

Palmophyllum crassum, a New Record of an Ancient Species in Green Algae from Korea

  • Lee, Hyung Woo;Kim, Myung Sook
    • 환경생물
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    • 제35권3호
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    • pp.319-328
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    • 2017
  • The continuous exploration in deep seawater from Korea makes us lead the discovery of ancient Chlorophyta, Palmophyllum, in the Korean coast. The phylogenetic analyses of 18S rRNA and rbcL genes demonstrate that our specimens are Palmophyllum crassum (Naccari) Rabenhorst, recorded in Japan and clearly distinguished from P. umbracola from New Zealand and California, USA. Palmophyllum crassum grows in the subtidal region, 8-30 m deep, and has a crustose thallus which is closely adherent to substrates such as non-geniculate crustose coralline algae, sponge, shells, or rocks. P. crassum is composed of numerous spherical cells embedded in the gelatinous matrix. The discovery of this ancient green seaweed implies that the Korean coast is one of the hotspots of algal species diversity and has the suitable marine environment for algal speciation. We suggest the grounds to conserve the Korean coast environmentally as the biodiversity center of marine species by studying the phylogeny of seaweeds.