Biofunctional Thermo-Responsive Polymeric Surface with Micropatterns for Label Free Cell Separation

2014 ◽  
Vol 1621 ◽  
pp. 107-112
Author(s):  
Yoshikazu Kumashiro ◽  
Jun Ishihara ◽  
Terumasa Umemoto ◽  
Kazuyoshi Itoga ◽  
Jun Kobayashi ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Umbilical Vein ◽  
Free Cell ◽  
Specific Cell ◽  
Label Free ◽  
Stripe Pattern ◽  
Adhesion Properties ◽  
Patterned Surface ◽  
Umbilical Vein Endothelial Cells ◽  
Response To Temperature

ABSTRACTThready stripe-patterned thermo-responsive surfaces were prepared and their surface properties were characterized. Prepared 3 μm wide stripe-patterned surfaces were evaluated by observing the adhesions and detachments of three types of cells: HeLa cells (HeLas), human umbilical vein endothelial cells (HUVECs), and NIH-3T3 cells (3T3s). Although cell adhesion and detachment in response to temperature were observed on all cells on a conventional thermo-responsive surface without patterns, the thermo-responsive surface with a 3 μm striped-pattern exhibited various cell adhesion properties. HeLas hardly adhered to the patterned surface even at 37 °C. On the other hand, although HUVECs adhered on the patterned surface at 12 h after incubation at 37 °C, the adhered HUVECs detached themselves after another 12 h incubation at 37 °C. 3T3s adhered to the patterned surface at 37 °C and detached themselves after reducing temperature to 20 °C. A mixture of HeLa, HUVEC and 3T3 was separated using their different specific cell-adhesion properties, and the composition of cells was analyzed by a flow-cytometry. As a result, the conventional thermo-responsive surface with a stripe-pattern was found to function as a cell-separating interface by using specific cell adhesion properties.

scholarly journals Mammalian and Drosophila cells adhere to the laminin α4 LG4 domain through syndecans, but not glypicans

2004 ◽  
Vol 382 (3) ◽  
pp. 933-943 ◽  
Author(s):  
Hironobu YAMASHITA ◽  
Akira GOTO ◽  
Tatsuhiko KADOWAKI ◽  
Yasuo KITAGAWA
Keyword(s):  
Cell Adhesion ◽  
Umbilical Vein ◽  
Heparan Sulphate ◽  
Direct Interaction ◽  
Binding Activity ◽  
Main Body ◽  
Heparin Binding ◽  
Adhesion Activity ◽  
Umbilical Vein Endothelial Cells

We have previously shown that the LG4 (laminin G-like) domain of the laminin α4 chain is responsible for the significantly higher affinity of the α4 chain to heparin than found for other α chains [Yamaguchi, Yamashita, Mori, Okazaki, Nomizu, Beck and Kitagawa (2000) J. Biol. Chem. 275, 29458–29465]; four basic residues were identified to be essential for this activity [Yamashita, Beck and Kitagawa (2004) J. Mol. Biol. 335, 1145–1149]. By creating GST (glutathione S-transferase)-fused LG1, LG2, LG4 and LG5 proteins, we found that only LG4 is active for the adhesion of human HT1080 cells, human umbilical vein endothelial cells and Drosophila haemocytes Kc167 with a half-saturating concentration of 20 μg/ml. Adhesion was counteracted by treatment of the cells with heparin, heparan sulphate and heparitinase I. Upon mutating the four basic residues essential for heparin binding within LG4, the adhesion activity was abolished. Pull-down experiments using glutathione beads/GST-fusion proteins indicate a direct interaction of LG4 with syndecan-4, which might be the major receptor for cell adhesion. Neither the release of glypican-1 by treating human cells with phosphatidylinositol-specific phospholipase C nor targeted knockdown of dally or dally-like protein impaired the cell-adhesion activity. As the LG4–LG5 domain of the α4 chain is cleaved in vivo from the main body of laminin-8 (α4β1γ1), we suggest that the heparan sulphate proteoglycan-binding activity of LG4 is significant in modulating the signalling of Wnt, Decapentaplegic and fibroblast growth factors.

scholarly journals Anti-Inflammatory and Anti-Apoptotic Effects of Stybenpropol A on Human Umbilical Vein Endothelial Cells

2019 ◽  
Vol 20 (21) ◽  
pp. 5383 ◽  
Author(s):  
Li Zhang ◽  
Feifei Wang ◽  
Qing Zhang ◽  
Qiuming Liang ◽  
Shumei Wang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Umbilical Vein ◽  
Caspase 9 ◽  
Protein Levels ◽  
Tnf Α ◽  
Umbilical Vein Endothelial Cells ◽  
Cell Adhesion Molecule 1

Inflammation is a key mediator in the progression of atherosclerosis (AS). Benzoinum, a resin secreted from the bark of Styrax tonkinensis, has been widely used as a form of traditional Chinese medicine in clinical settings to enhance cardiovascular function, but the active components of the resin responsible for those pharmaceutical effects remain unclear. To better clarify these components, a new phenylpropane derivative termed stybenpropol A was isolated from benzoinum and characterized via comprehensive spectra a nalysis. We further assessed how this phenylpropane derivative affected treatment of human umbilical vein endothelial cells (HUVECs) with tumor necrosis factor-α (TNF-α). Our results revealed that stybenpropol A reduced soluble intercellular cell adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), interleukin-8 (IL-8), and interleukin-1β (IL-1β) expression by ELISA, inhibited apoptosis, and accelerated nitric oxide (NO) release in TNF-α-treated HUVECs. We further found that stybenpropol A decreased VCAM-1, ICAM-1, Bax, and caspase-9 protein levels, and increased the protein levels of Bcl-2, IKK-β, and IκB-α. This study identified a new, natural phenylpropane derivative of benzoinum, and is the first to reveal its cytoprotective effects in the context of TNF-α-treated HUVECs via regulation of the NF-κB and caspase-9 signaling pathways.

Abstract 110: Thrombospondin Type-1 Domain-Containing Protein 1 (THSD1), an Intracranial Aneurysm Susceptibility Gene, Mediates Cell Adhesion in Human Umbilical Vein Endothelial Cells

Stroke ◽  
2015 ◽  
Vol 46 (suppl_1) ◽  
Author(s):  
Xiaoqian Fang ◽  
Dong H Kim ◽  
Teresa Santiago-Sim
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Focal Adhesion ◽  
Umbilical Vein ◽  
Adhesion Formation ◽  
Wild Type ◽  
Focal Adhesion Formation ◽  
Umbilical Vein Endothelial Cells

Introduction: An intracranial aneurysm (IA) is a weak spot in cerebral blood vessel wall that can lead to its abnormal bulging. Previously, we reported that mutations in THSD1 , encoding thrombospondin type-1 domain-containing protein 1, are associated with IA in a subset of patients. THSD1 is a transmembrane molecule with a thrombospondin type-1 repeat (TSR). Proteins with TSR domain have been implicated in a variety of processes including regulation of matrix organization, cell adhesion and migration. We have shown that in mouse brain Thsd1 is expressed in endothelial cells. Hypothesis: THSD1 plays an important role in maintaining the integrity of the endothelium by promoting adhesion of endothelial cells to the underlying basement membrane. Methods: Human umbilical vein endothelial cells are used to investigate the role of THSD1 in vitro . THSD1 expression was knocked-down by RNA interference. Cell adhesion assay was done on collagen I-coated plates and focal adhesion formation was visualized using immunofluorescence by paxillin and phosphorylated focal adhesion kinase (pFAK) staining. THSD1 re-expression is accomplished by transfection with a pCR3.1-THSD1-encoding plasmid. Results: Knockdown of THSD1 caused striking change in cell morphology and size. Compared to control siRNA-treated cells that exhibited typical cobblestone morphology, THSD1 knockdown cells were narrow and elongated, and were significantly smaller ( p <0.01). Cell adherence to collagen I-coated plates was also attenuated in THSD1 knockdown cells ( p <0.01). Consistent with this finding is the observation that the number and size of focal adhesions, based on paxillin and pFAK staining, were significantly reduced after THSD1 knockdown ( p <0.01). These defects in cell adhesion and focal adhesion formation were rescued by re-expression of wild type THSD1 ( p <0.05). In contrast, initial studies indicate that expression of mutated versions of THSD1 as seen in human patients (L5F, R450*, E466G, P639L) could not restore cell adhesion and focal adhesion formation to wild type levels. Conclusions: Our studies provide evidence for a role of THSD1 and THSD1 mutations in endothelial cell adhesion and suggest a possible mechanism underlying THSD1 -mediated aneurysm disease.

scholarly journals Microcystins Induces Vascular Inflammation in Human Umbilical Vein Endothelial Cells via Activation of NF-κB

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Jun Shi ◽  
Jie Zhou ◽  
Min Zhang
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Adhesion Molecules ◽  
Cell Adhesion Molecules ◽  
Inflammatory Process ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Health Hazard ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Microcystins (MCs) produced by toxic cyanobacteria cause serious water pollution and public health hazard to humans and animals. However, direct molecular mechanisms of MC-LR in vascular endothelial cells (ECs) have not been understood yet. In this study, we investigated whether MC-LR induces vascular inflammatory process in cultured human umbilical vein endothelial cells (HUVECs). Our data demonstrated that MC-LR decreased HUVECs proliferation and tube formation and enhanced apoptosis. MC-LR also induced intracellular reactive oxygen species formation (ROS) in HUVECs. The MC-LR directly stimulated phosphorylation of NF-κB. Furthermore, MC-LR also increased cell adhesion molecules (ICAM-1 and VCAM-1) expression in HUVECs. Taken together, the present data suggested that MC-LR induced vascular inflammatory process, which may be closely related to the oxidative stress, NF-κB activation, and cell adhesion molecules expression in HUVECs. Our findings may highlight that MC-LR causes potential damage to blood vessels.

scholarly journals Ellagic acid inhibits IL-1β-induced cell adhesion molecule expression in human umbilical vein endothelial cells

2007 ◽  
Vol 97 (4) ◽  
pp. 692-698 ◽  
Author(s):  
Ya-Mei Yu ◽  
Zhi-Hong Wang ◽  
Chung-Hsien Liu ◽  
Chin-Seng Chen
Keyword(s):  
Reactive Oxygen Species ◽  
Endothelial Cells ◽  
Cell Adhesion ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Nuclear Translocation ◽  
Umbilical Vein ◽  
Oxygen Species ◽  
Adhesion Molecule Expression ◽  
Umbilical Vein Endothelial Cells

Expression of cell adhesion molecules by endothelium and the attachment of monocytes to endothelium may play a major role in atherosclerosis. Ellagic acid (EA) is a phenolic compound found in fruits and nuts including raspberries, strawberries, grapes and walnuts. Previous studies have indicated that EA possesses antioxidant activity in vitro. In the present study, we investigated the effects of EA on the formation of intracellular reactive oxygen species, the translocation of NFκB and expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 and endothelial leucocyte adhesion molecule (E-selectin) induced by IL-1β in human umbilical vein endothelial cells (HUVEC). We found that EA significantly reduced the binding of human monocytic cell line, U937, to IL-1β-treated HUVEC. The production of reactive oxygen species by IL-1β was dose-dependently suppressed by EA. Supplementation with increasing doses of EA up to 50 μmol/l was most effective in inhibiting the expression of VCAM-1 and E-selectin. Furthermore, the inhibition of IL-1β-induced adhesion molecule expression by EA was manifested by the suppression of nuclear translocation of p65 and p50. In conclusion, EA inhibits IL-1β-induced nuclear translocation of p65 and p50, thereby suppressing the expression of VCAM-1 and E-selectin, resulting in decreased monocyte adhesion. Thus, EA has anti-inflammatory properties and may play an important role in the prevention of atherosclerosis.

scholarly journals Synthesis of 1,25-Dihydroxyvitamin D3 by Human Endothelial Cells Is Regulated by Inflammatory Cytokines: A Novel Autocrine Determinant of Vascular Cell Adhesion

2002 ◽  
Vol 13 (3) ◽  
pp. 621-629
Author(s):  
Daniel Zehnder ◽  
Rosemary Bland ◽  
Ravinder S. Chana ◽  
David C. Wheeler ◽  
Alexander J. Howie ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Inflammatory Cytokines ◽  
Vascular Tissue ◽  
Umbilical Vein ◽  
Primary Cultures ◽  
Human Endothelial Cells ◽  
Functional Studies ◽  
Reverse Transcription Pcr ◽  
Umbilical Vein Endothelial Cells

ABSTRACT. In addition to its calciotropic function, the secosteroid 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) has potent nonclassical effects. In particular, local production of 1,25D3 catalyzed by the enzyme 1α-hydroxylase (1α-OHase) may act as an autocrine/paracrine immunomodulatory mechanism. To investigate the significance of this in vascular tissue the expression and function of 1α-OHase in human endothelial cells was characterized. Immunohistochemical and in situ hybridization analyses show, for the first time, the presence of 1α-OHase mRNA and protein in endothelial cells from human renal arteries as well as postcapillary venules from lymphoid tissue. Reverse transcription–PCR and Western blot analyses confirmed the presence of 1α-OHase in primary cultures of human umbilical vein endothelial cells (HUVEC). Enzyme activity in HUVEC (318 ± 56 fmoles 1,25(OH)2D3/hr/mg protein) increased after treatment with tumor necrosis factor–α (1054 ± 166, P < 0.01), lipopolysaccharide (1381 ± 88, P < 0.01), or forskolin (554 ± 56, P < 0.05). Functional studies showed that exogenously added 1,25(OH)2D3 or its precursor, 25-hydroxyvitamin D3 (25(OH)D3), significantly decreased HUVEC proliferation after 72 h of treatment (33% and 11%, respectively). In addition, after 24 h treatment, both 1,25(OH)2D3 and 25(OH)D3 increased the adhesion of monocytic U937 cells to HUVEC (159% and 153%, respectively). These data indicate that human endothelia are able to produce active vitamin D. The rapid induction of endothelial 1α-OHase activity by inflammatory cytokines suggests a novel autocrine/paracrine role for the enzyme, possibly as a modulator of endothelial cell adhesion.

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