Osteopontin (OPN) is an extracellular glycosylated phosphoprotein that promotes cell adhesion by interacting with several integrin receptors. We previously reported that an OPN mutant lacking five O-glycosylation sites (Thr134/Thr138/Thr143/Thr147/Thr152) in the threonine/proline-rich region increased cell adhesion activity and phosphorylation compared with the wild type. However, the role of O-glycosylation in cell adhesion activity and phosphorylation of OPN remains to be clarified. Here, we show that site-specific O-glycosylation in the threonine/proline-rich region of OPN affects its cell adhesion activity and phosphorylation independently and/or synergistically. Using site-directed mutagenesis, we found that OPN mutants with substitution sets of Thr134/Thr138 or Thr143/Thr147/Thr152 had decreased and increased cell adhesion activity, respectively. In contrast, the introduction of a single mutation into the O-glycosylation sites had no effect on OPN cell adhesion activity. An adhesion assay ...
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This application note analyzes the role of different adhesion molecules and chemokines involved in various stages of inflammation under physiological flow conditions. Using Cellixs biochips and Mirus pumping system, THP-1, monocyte and PBMC adhesion to VCAM-1; THP-1, monocyte and PBMC rolling on E-selectin; and respective adhesion blockades is investigated. THP-1 adhesion to HUVECs, correlating adhesion assay results with adhesion molecule expression levels on HUVECs from flow cytometry data, i
Expression of cyclooxygenases (COX) and lipoxygenases (LOX) has been linked to many pathophysiological phenotypes, including cell adhesion. However, many current approaches to measure cellular changes are performed only in a fixed-time point. Since cells dynamically move in conjunction with the cell matrix, there is a pressing need for dynamic or time-dependent methods for the investigation of cell properties. In the presented study, we used stable human colorectal cancer cell lines ectopically expressing COX-1, COX-2, and 15LOX-1, to investigate whether expression of COX-1, COX-2, or 15LOX-1 would affect cell adhesion using our opto-electric methodology. In a fixed-time point experiment, only COX-1- and COX-2-expressing cells enhanced phosphorylation of focal adhesion kinase, but all the transfected cells showed invasion activity. However, in a real-time experiment using opto-electric approaches, transmitted cellular morphology was much different with tight adhesion being shown in COX-2 expressing
The extravasation of leukocytes from the blood into tissues occurs as a multistep process: an initial transient interaction (rolling), generally thought to be mediated by the selectin family of adhesion molecules, followed by firm adhesion, usually mediated by integrins. Using a parallel plate flow chamber designed to approximate physiologic flow in postcapillary venules, we have characterized a rolling interaction between lymphoid cells and adherent primary and cultured endothelial cells that is not selectin mediated. Studies using blocking monoclonal antibodies indicate that this novel interaction is mediated by CD44. Abrogation of the rolling interaction could be specifically achieved using both soluble hyaluronate (HA) and treatment of the adherent cells with HA-reactive substances, indicating that HA is the ligand supporting this rolling interaction. Some B and T cell lines, as well as normal lymphocytes, either constitutively exhibit rolling or can be induced to do so by phorbol ester or ...
Cell adhesion involves receptor-mediated cell-surface interactions with the extracellular matrix (Burridge and Chrzanowska-Wodnicka 1996; Gumbiner 1996). These interactions play a central role in the organization of the cytoskeleton, thereby regulating cell shape and function. Focal adhesions are specialized structures linking the extracellular matrix to the actin microfilaments through integrin and syndecan transmembrane receptors. The structure of the focal adhesion plaque consists of an elaborate network of interconnecting proteins anchoring the microfilaments to the membrane at the contact site. As the points of closest apposition linking the cytoskeleton to the extracellular matrix, focal adhesions are ideally positioned for regulating the adhesive strength of the cell. It may help to think of the cell as having three grades of adhesiveness: (1) weak adherence, meaning that the cell is attached but not spread; (2) intermediate adherence, characterized by a spread cell that lacks stress ...
L-Arginine reduces human monocyte adhesion to endothelial cells and decreases expression of certain endothelial cell adhesion molecules.
The integrin LFA-1 and its ligand ICAM-1 mediate B cell adhesion, but their role in membrane-bound antigen recognition is still unknown. Here, using planar lipid bilayers and cells expressing ICAM-1 fused to green fluorescence protein, we found that the engagement of B cell receptor (BCR) promotes B cell adhesion by an LFA-1-mediated mechanism. LFA-1 is recruited to form a mature B cell synapse segregating into a ring around the BCR. This distribution is maintained over a wide range of BCR/antigen affinities (10(6) M(-1) to 10(11) M(-1)). Furthermore, the LFA-1 binding to ICAM-1 reduces the level of antigen required to form the synapse and trigger a B cell. Thus, LFA-1/ICAM-1 interaction lowers the threshold for B cell activation by promoting B cell adhesion and synapse formation.
The aim of the first part of the thesis was to develop and validate an in vitro adherence assay involving porcine mononuclear cells (MCs) and porcine endothelium, present within gut and lymph node. Factors involved in MC / endothelium interactions were determined. In summary we found that cell adhesion in our assay system was temperature, Ca2+ and Mn2+ sensitive, required metabolic activity, was inhibited by the phosphorylated monosaccharide galactose 6-phosphate, and unaffected by the presence of mucus. These findings reflected certain aspects of in vivo cell adhesion, present within the in vitro assay used. The adhesion characteristics of porcine Peyer s patch (PP), peripheral blood (PB), and lymph node (LN) MCs to porcine gut and lymph node endothelium was examined and used as an guiding model for the future study of human MCs adherence. It was found that PP MCs adhered significantly better to gut endothelium than to LN endothelium and similarly LN MCs adhered significantly better to LN ...
Cell adhesion to extracellular matrix (ECM) is critical to various cellular processes like cell spreading, migration, growth and apoptosis. At the tissue level, cell adhesion is important in the pathological and physiological processes that regulate the tissue morphogenesis. Cell adhesion to the ECM is primarily mediated by the integrin family of receptors. The receptors that are recruited to the surface are reinforced by structural and signaling proteins at the adhesive sites forming focal adhesions that connect the cytoskeleton to further stabilize the adhesions. The functional roles of these focal adhesions extend beyond stabilizing adhesions and transduce mechanical signals at the cell-ECM interface in various signaling events. The objective of this research is to analyze the role of the spatial distribution of the focal adhesions in stabilizing the cell adhesion to the ECM in relation to cells internal force balance. The central hypothesis was that peripheral focal adhesions stabilize cell
Static adhesion of transfectants to immobilized ligands. Adhesion of various integrin transfectants to MAdCAM-1 (top) and ICAM-1 (bottom) was measured in
The modification of medical device surface with adhesive ligands has been recently shown to be an effective means for making a bioselective surface which can inhibit bacterial adhesion while promoting host cell adhesion on device materials. Currently, the lack of quantitative correlation between the adhesion strength of bacteria, nature of adhesive ligand and adhesion kinetics of mammalian cells hinders the development of such device surface. In this study, the biophysical responses of bacteria and mammalian cells towards adhesive ligand on model device surfaces formed by the chemisorption of dopamine (a moderate antibiotic) on glass are elucidated. The effects of RGD, collagen and dopamine modification on the adhesion strength of two clinically significant bacteria including Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were investigated by the determination of minimum lateral forces for bacterial detachment and the density of adhering bacteria. The result indicates that RGD ...
Antioxidants have been proposed to be anti-atherosclerotic agents; however, the mechanisms underlying their beneficial effects are poorly understood. We have examined the effect of alpha-tocopherol (alpha-tcp) on one cellular event in atherosclerotic plaque development, monocyte adhesion to stimulated endothelial cells (ECs). Human umbilical vein ECs were pretreated with alpha-tcp before stimulation with known agonists of monocyte adhesion: IL-1 (10 ng/ml), LPS (10 ng/ml), thrombin (30 U/ml), or PMA (10 nM). Agonist-induced monocytic cell adhesion, but not basal adhesion, was inhibited in a time- and concentration-dependent manner by alpha-tcp. The IC50 of alpha-tcp on an IL-1-induced response was 45 microM. The inhibition correlated with a decrease in steady state levels of E-selectin mRNA and cell surface expression of E-selectin which is consistent with the ability of a monoclonal antibody to E-selectin to inhibit monocytic cell adhesion in this system. Probucol (50 microM) and ...
In the 9 years since the last review on leukocyte and endothelial interactions was published in this journal many of the critical structures involved in leukocyte adherence to and migration across endothelium have been elucidated. With the advent of cell and molecular biology approaches, investigations have progressed from the early descriptions by intravital microscopy and histology, to functional and immunologic characterization of adhesion molecules, and now to the development of genetically deficient animals and the first phase I trial of anti-adhesion therapy in humans. The molecular cloning and definition of the adhesive functions of the leukocyte integrins, endothelial members of the Ig gene superfamily, and the selectins has already provided sufficient information to construct an operative paradigm of the molecular basis of leukocyte emigration. The regulation of these adhesion molecules by chemoattractants, cytokines, or chemokines, and the interrelationships of adhesion pathways need ...
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The integrin subunit beta 1B, a beta 1 isoform with a unique sequence at the cytoplasmic domain, forms heterodimers with integrin alpha chains and binds fibronectin, but it does not localize to focal adhesion sites (Balzac, F., A. Belkin, V. Koteliansky, Y. Balabanow, F. Altruda, L. Silengo, and G. Tarone. 1993. J. Cell Biol. 121:171-178). Here we analyze the functional properties of human beta 1B by expressing it in hamster CHO cells. When stimulated by specific antibodies, beta 1B does not trigger tyrosine phosphorylation of a 125-kD cytosolic protein, an intracellular signalling pathway that is activated both by the endogenous hamster or the transfected human beta 1A. Moreover, expression of beta 1B results in reduced spreading on fibronectin and laminin, but not on vitronectin. Expression of beta 1B also results in severe reduction of cell motility in the Boyden chamber assay. Reduced cell spreading and motility could not be accounted for by preferential association of beta 1B with a given ...
Cell adhesion to the extracellular matrix is required to execute growth factor (GF)-mediated cell behaviors, such as proliferation. A major underlying mechanism is that cell adhesion enhances GF-mediated intracellular signals, such as extracellular signal-regulated kinase (Erk). However, because GFs use distinct mechanisms to activate Ras-Erk signaling, it is unclear whether adhesion-mediated enhancement of Erk signaling is universal to all GFs. We examined this issue by quantifying the dynamics of Erk signaling induced by epidermal growth factor, basic fibroblast growth factor (bFGF), and platelet-derived growth factor (PDGF) in NIH-3T3 fibroblasts. Adhesion to fibronectin-coated surfaces enhances Erk signaling elicited by epidermal growth factor but not by bFGF or PDGF. Unexpectedly, adhesion is not always a positive influence on GF-mediated signaling. At critical subsaturating doses of PDGF or bFGF, cell adhesion ablates Erk signaling; that is, adhesion desensitizes the cell to GF ...
Cell adhesion is a fundamental phenomenon vital for all multicellular organisms. Recognition of and adhesion to specific macromolecules is a crucial task of leukocytes to initiate the immune response. To gain statistically reliable information of cell adhesion, large numbers of cells should be measured. However, direct measurement of the adhesion force of single cells is still challenging and todays techniques typically have an extremely low throughput (5-10 cells per day). Here, we introduce a computer controlled micropipette mounted onto a normal inverted microscope for probing single cell interactions with specific macromolecules. We calculated the estimated hydrodynamic lifting force acting on target cells by the numerical simulation of the flow at the micropipette tip. The adhesion force of surface attached cells could be accurately probed by repeating the pick-up process with increasing vacuum applied in the pipette positioned above the cell under investigation. Using the introduced methodology
Hematogenous metastasis requires the arrest and extravasation of blood-borne tumor cells, possibly involving direct adhesive interactions with vascular endothelium. Cytokine activation of cultured human endothelium increases adhesion of melanoma and carcinoma cell lines. An inducible 110-kD endothelial cell surface glycoprotein, designated INCAM-110, appears to mediate adhesion of melanoma cells. In addition, an inducible endothelial receptor for neutrophils, ELAM-1, supports the adhesion of a human colon carcinoma cell line. Thus, activation of vascular endothelium in vivo that results in increased expression of INCAM-110 and ELAM-1 may promote tumor cell adhesion and affect the incidence and distribution of metastases.. ...
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TY - JOUR. T1 - Control of integrin αIIbβ3 outside-in signaling and platelet adhesion by sensing the physical properties of fibrin(ogen) substrates. AU - Podolnikova, Nataly. AU - Yermolenko, Ivan S.. AU - Fuhrmann, Alexander. AU - Lishko, Valeryi K.. AU - Magonov, Sergei. AU - Bowen, Benjamin. AU - Enderlein, Joerg. AU - Podolnikov, Andriy V.. AU - Ros, Robert. AU - Ugarova, Tatiana. PY - 2010/1/12. Y1 - 2010/1/12. N2 - The physical properties of substrates are known to control cell adhesion via integrin-mediated signaling. Fibrin and fibrinogen, the principal components of hemostatic and pathological thrombi, may represent biologically relevant substrates whose variable physical properties control adhesion of leukocytes and platelets. In our previous work, we have shown that binding of fibrinogen to the surface of fibrin clot prevents cell adhesion by creating an antiadhesive fibrinogen layer. Furthermore, fibrinogen immobilized on various surfaces at high density supports weak cell adhesion ...
The broad tissue distribution and evolutionary conservation of the GPI-anchored protein PrP suggests that it plays a role in cellular homeostasis. Since integrin adhesion determines cell behavior, the proposed role of PrP in cell adhesion may underlie the various in vitro and in vivo effects associated to PrP loss-of-function, including the immune phenotypes described in PrP−/- mice. We have investigated the role of PrP in the adhesion and (transendothelial) migration of human (pro)monocytes. We found that PrP regulates β1 integrin-mediated adhesion of monocytes. Additionally, PrP controls cell morphology and migratory behavior of monocytes: PrP-silenced cells show deficient uropod formation on immobilized VCAM and display bleb-like protrusions on the endothelium. Our data further show that PrP regulates ligand-induced integrin activation. Finally, we found that PrP controls the activation of several proteins involved in cell adhesion and migration, including RhoA and its effector cofilin as ...
University of Turku. Professor Johanna Ivaskas (University of Turku) research focus is on the changes that occur in cells with the development of cancer metastases. Integrins are important cell adhesion receptors that regulate the division of cells and their movement in tissue. Changes in cell adhesion properties are a key factor in the formation of cancer metastases. The aim of Professor Ivaskas research is to reach a fundamentally new mechanical understanding of how integrins work in cancer cells and to produce a roadmap of integrin receptor operation and communication chains.. The research combines different methods, including in vivo models, high-throughput screening and applications of synthetic biology. These innovative approaches will yield significant new information about the pathways of cancer cells and their movement in tissue. The project is expected to result in major scientific breakthroughs in this topical field of biomedicine.. Johanna Ivaska is a highly merited researcher. For ...
The idea that cells adhere to one another in a specific manner, such that cells of one type stick only to cells of the same type, appears to have had its origin from the work of Wilson (1907). He found that when cell suspensions from two species of marine sponge were mixed and allowed to aggregate, each individual aggregate body was composed of cells of one species alone. This conclusion has been supported by the results obtained by Humphreys (1963) amongst others, though some workers, who have used different species of sponge, have failed to detect signs of specific adhesion of the cells (Sara, Liaci & Melone, 1966). Until recently there has been little evidence in favour or against the idea that specific adhesion occurs between the cells of higher animals.. ...
Project leader: Prof. Dr. T. Chavakis. Integrin-dependent adhesive interactions between leukocytes and the endothelium contribute to inflammatory processes. In addition, similar adhesive events between haematopoietic stem cells (HSC) and bone marrow stromal cells, including endothelial cells, play a major role for the mobilisation of HSC into peripheral blood and for the homing of HSC to the bone marrow, both processes being relevant for bone marrow transplantation. The beta2-integrin LFA-1, exclusively expressed on cells of haematopoietic origin, is a major adhesion receptor in this context. We recently identified developmental endothelial locus-1 (Del-1 or Edil3), secreted by endothelial cells, as an endogenous inhibitor of LFA-1-dependent leukocyte adhesion to endothelial cells in vitro and leukocyte recruitment in vivo as well as of interleukin-17 (IL-17)-dependent inflammation in the context of aging-associated inflammatory bone loss. Since LFA-1 can regulate adhesive functions of HSC and ...
The invention discloses a cell culture support which provides for the adhesion and culturing of one or more adhesive cells using a photoresist in which to provide a particular patterned design on a surface of the support. The patterned design is provided by the photoresist which is partially removed by photolithography during the making of the support which in turn imparts a striped, checkerboard or dotted pattern on the surface of the support. Further, the cell culture support is produced by pretreating the support surface with a reagent to provide hydrophobicity to the support surface. Also a reagent can be added to pretreat the support surface in order to facilitate adhesion at the photoresist prior to applying the photoresist into the cell culture support. Collagen is applied in the form of a solution, containing in addition thereto albumin and a crosslinking agent, in order to form a film. Collagen specifically affects the cell adhesion rate or the morphology of the cells to be adhered to the
The Phd thesis titled The role of trade partners cohesiveness in the conclusion of interregional agreements with the European Union explores the most important factors helping cohesiveness to explain the likelihood of concluding an agreement with the EU.
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Cells exert actomyosin contractility and cytoskeleton-dependent force in response to matrix stiffness cues. Cells dynamically adapt to force by modifying their behavior and remodeling their microenvironment. This adaptation is favored by integrin activation switch and their ability to modulate their clustering and the assembly of an intracellular hub in response to force. Indeed integrins are mechanoreceptors and mediate mechanotransduction by transferring forces to specific adhesion proteins into focal adhesions which are sensitive to tension and activate intracellular signals. α(5)β(1) integrin is considered of major importance for the formation of an elaborate meshwork of fibronectin fibrils and for the extracellular matrix deposition and remodeling. Here we summarize recent progress in the study of mechanisms regulating the activation cycle of β(1) integrin and the specificity of α(5)β(1) integrin in mechanotransduction.
Nano-scale or micro-scale adhesive structures comprising an array of nano-fabricated, pillars, the pillars having coated upon, or having disposed on a working surface thereof, a protein-mimetic, marine-adhesive coating. Methods of fabricating the nano-scale pillars, synthesis of the protein-mimetic coating or wet adhesive and application of the adhesive to the pillars are described.
The functionalization of hydrogels for receptor-mediated cell adhesion is one approach for targeted cell and tissue engineering applications. In this study, polyacrylamide gel surfaces were functionalized with specific cell adhesion ligands via the self-assembly of a peptide-based heterodimer. The s …
The CD2 receptor on T lymphocytes is essential for T cell adhesion and stimulation by antigen presenting cells (APCs). Blockade of CD2 function is immunosuppressive in both model systems and humans, indicating the importance of CD2 for the cellular immune response. Although the affinity of the molecular interaction between CD2 and its counter-receptor, CD58, is relatively low when measured in solution, this interaction mediates tight adhesion within the 2D cell-cell interface. To understand the mechanisms responsible for regulating the avidity of the CD2-CD58 interaction, we measured the number, affinity, and lateral mobility of CD2 molecules on resting and activated T cells. Cell activation caused a 1.5-fold increase in the number of CD2 sites on the cell surface, and the 2D affinity of CD2 for CD58 increased by 2.5-fold. The combination of T cell activation and CD2 ligation to CD58 decreased the laterally mobile fraction of the ligated CD2. Together, these changes would substantially enhance CD2
The integrin VLA-3 is a cell surface receptor, which binds to fibronectin, laminin, collagen type I and VI (Takada, Y., E. A. Wayner, W. G. Carter, and M. E. Hemler. 1988. J. Cell. Biochem. 37:385-393) and is highly expressed in substrate adherent cultures of almost all human cell types. The ligand specificity of VLA-3 and the inhibition of cell adhesion by anti-VLA-3 monoclonal antibodies suggest its involvement in cell-substrate interaction. In normal tissues, VLA-3 is restricted to few cell types, notably the kidney glomeruli and basal cells of the epidermis. In the epidermis, VLA-3 is generally strongly expressed on the entire plasma membrane of basal cells and is not polarized towards the basement membrane (Klein, C. E., C. Cardon-Cardo, R. Soehnchen, R. J. Cote, H. F. Oettgen, M. Eisinger, and L. J. Old. 1987. J. Invest. Dermatol. 89:500-507). Based on this finding we speculated that, in addition to a role of VLA-3 for adhesion of cells to substrate, it could also be relevant for cell-cell ...
Colorectal tumors originate and develop within intestinal crypts. Even though some of the essential phenomena that characterize crypt structure and dynamics have been effectively described in the past, the relation between the differentiation process and the overall crypt homeostasis is still partially understood. We here investigate this relation and other important biological phenomena by introducing a novel multiscale model that combines a morphological description of the crypt with a gene regulation model: the emergent dynamical behavior of the underlying gene regulatory network drives cell growth and differentiation processes, linking the two distinct spatio-temporal levels. The model relies on a few a priori assumptions, yet accounting for several key processes related to crypt functioning, such as: dynamic gene activation patterns, stochastic differentiation, signaling pathways ruling cell adhesion properties, cell displacement, cell growth, mitosis, apoptosis and the presence of ...
The role of the mesothelial layer in the peritoneal spreading of cancer cells is only partially clarified. Here we attempted to better define the mesothelial contribution to the tumor cell adhesion using a direct adhesion test applied to human primary cultures of mesothelial cells (HPMCs) derived from the peritoneal washes of patients with gastric and colorectal cancers. Gastric and colon carcinoma cells were seeded on different mesothelial monolayers and quantitative fluorescence analysis was performed to analyze their growth and adhesive properties. The adhesion of the cancer cells was not affected by the origin of the HPMCs when derived from patients with different cancers or with benign disease. In contrast, the high levels of ICAM1 expression and ROS production, which characterize these senescent mesothelial cells, enhanced the tumor cell adhesion. These results suggest that the mesothelial adhesive properties are dependent on the cell senescence, while are not affected by the tumor ...
My recent research has focused on how leukocytes control their adhesiveness and has resulted in a discovery of a novel regulatory pathway, Rogelj relates. Its a Rube Goldberg kind of sequence: Biochemical events that occur along this pathway determine the expression of a critical cell surface adhesion molecule, which in turn determines the ability of a leukocyte to recognize its target. Signals that lead to a loss of cell adhesiveness, therefore, result in suppression of the immune response. And, suppression of the immune system may or may not be a good thing ...
Our laboratory consists of 5 Research Fellows and a Junior Faculty member who are physician scientists or research scientists and two senior research technicians. who use a combination of immunological, biochemical and molecular biological strategies to study leukocyte recruitment in various in vitro and in vivo models of inflammation. We have developed a valuable in vitro model that allows direct microscopic examination of live leukocyte Ð endothelial interactions under defined laminar fluid shear stress conditions that mimic blood flow in small venules. Areas of focus using this model are three-fold: first, dissection of the adhesion mechanisms that support blood monocyte and specific T cell subset adhesive interactions with endothelium under flow, or specific recombinant endothelial cell adhesion molecules; second, characterization of endothelial-dependent mechanisms involved in regulation of endothelial cell borders (lateral junctions) during leukocyte transmigration, permeability function ...
The Ly-6 locus on mouse chromosome 15 encodes a family of 10-12 kDa proteins that are linked to the cell surface by a glycosylphosphatidyl-inositol anchor and have cell signaling and cell adhesion properties. Expression of Ly-6 proteins is tightly regulated during development; these proteins continu.... Full description. ...
The leukocyte adhesion cascade is an important paradigm of immunity and mediates leukocyte recruitment in acute or chronic inflammatory responses. Leukocyte recruitment requires several adhesive interactions between leukocytes and endothelial cells. The adhesion of leukocytes to the endothelial cell surface is mediated by interactions between leukocyte integrins, such as the beta1-integrin family member VLA-4 (a4b1) or the beta2-integrin family members LFA-1 (aLb2, CD11a/CD18), Mac-1 (aMb2, CD11b/CD18, complement receptor-3), and their endothelial counter-receptors of the immunoglobulin superfamily (ICAM-1, VCAM-1) (1). Our lab has made significant contributions to the leukocyte adhesion cascade, including the recent identification of a novel endogenous inhibitor of leukocyte recruitment, the endothelial-derived molecule Developmental Endothelial Locus-1 (Del-1, Edil3) (2-4).. Mobilization of hematopoietic stem cells (HSC) from the bone marrow to the periphery takes place upon infection. HSC ...
Prostate Cancer (PCa) is the second leading cause of cancer death in American men.. The inflammatory tumor microenvironment is a fertile niche that releases reactive oxygen species, which accelerates the malignant transformation and appears as a fine tuner of the adhesive behavior of cells. Heme oxygenase 1 (HO-1), the rate-limiting enzyme in heme degradation, represents an essential event in cellular responses to pro-oxidative and pro-inflammatory insults. As we previously reported that HO-1 over-expression impaired tumor growth and angiogenesis in vivo we sought to assess whether HO-1 could regulate the adhesive properties and the morphology of PCa cells. A bioinformatics enrichment analysis using Metacore, GeneMANIA and DAVID was performed; rendering a significant association of the HO-1 regulated genes with several proteins located in the extracellular space and cell membrane; compartments highly correlated with the adhesive behavior of cells. In an effort to understand the molecular ...
Effect of Ni addition on the microstructures of melt-spun CuCr ribbons. YU, M.; WANGI, Y.; WANG, Y.; SUN, Z. // Materials Science (0137-1339);2008, Vol. 26 Issue 3, p675 The microstructures and resistivities of melt-spun Cu75Cr25 and Cu(75-x)Cr25Nix (x = 1 or 3 wt. %) ribbons were studied. The size of the Cr-rich phase from liquid phase separation in the Cu75Cr25 microstructure can be decreased from the micrometer-scale to about 250 nm by using melt spinning.... ...
Sample preparation has always been a notoriously time consuming task that tends to detract from the more essential functions of collecting and analyzing data. A few key factors come in to play with improving the process of plating cells for imaging and the time that it takes. Cell adhesion, media re-equilibration, an unobstructed path for free migration of cells are some of the important factors to improve plating efficiency. Cell adhesion is pendant on the surface the cells are being plated on. The chemical composition of the glass affects cell adhesion and all glass surfaces are not created equal. It is best to use glass that is alkaline free and designed for cell adhesion (check out the Bioptechs Delta T Culture Dishes, FCS2 coverslips, 30mm ICD coverslips, and Microaquaduct slides). Sometimes an ECM is required depending on the cell type and protocol, however, in all cases cell plating is improved with the use of Culture Cylinders. A unique attribute of using a Culture Cylinder for plating ...
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Here, we investigated molecular and biophysical mechanisms through which GBM cells adhere to HA matrix. We find that GBM cells interact with HA via McTNs, which are long, thin protrusions rich in the HA receptor CD44, which we show is necessary for cell adhesion and McTN stability. Motility assays and laser ablation indicate that McTNs are stabilized by a balance of actomyosin-driven contraction, microtubule-driven protrusion, and CD44-mediated adhesion. Mechanical coupling of actin and microtubules with McTNs appears to be reinforced by a complex of IQGAP1 and CLIP170, and loss of IQGAP1 specifically disrupts adhesion and motility on HA.. McTNs, which can be functionally defined as microtubule-positive protrusions that elongate following actin depolymerization, have been observed in circulating tumor cells (CTCs), where they are believed to facilitate endothelial adhesion prior to extravasation (30⇓-32). However, to our knowledge, these structures have not been previously appreciated in the ...
Cell adhesion to the extracellular matrix (ECM) is necessary for fundamental cellular processes such as survival, migration, and differentiation. Adhesion is mediated by integrin receptors, which recruit multiprotein adhesion complexes to sites of attachment to the ECM. Adhesion complexes provide a structural connection between the ECM and cytoskeleton, transmit mechanical force, and act as signaling hubs to control cell behavior. Recent high-resolution imaging studies of adhesion sites reveal some aspects of their spatial organization and provide insights into their function at the molecular level.. ...
In this study, we have investigated whether SHIP plays a role in PMA- or cytokine-mediated LFA-1 activation by overexpressing both WT and phosphatase dead forms of SHIP in DA-ER cells. Our results show that 1) overexpression of WT-SHIP in unstimulated DA-ER cells increases LFA-1-mediated cell adhesion to ICAM-1, and this adhesion is further augmented by the addition of PMA, IL-3, or Epo; 2) SHIP requires a functional 5′-phosphatase domain for these effects, and overexpression of a phosphatase dead form actually leads to a slight inhibition of LFA-1-mediated adhesion to ICAM-1; 3) SHIP overexpression most likely enhances adhesion via its effect on inside-out signaling because its overexpression has no effect on the external activation of LFA-1 by Mn2+; 4) LFA-1 activation on cells overexpressing WT-SHIP does not involve activation of Erk-1 and Erk-2; and 5) LFA-1 activation in response to PMA in SHIP-overexpressing cells is via its effects on a PKC-stimulated pathway, while LFA-1 activation in ...
We have designed a lightly crosslinked PEG based copolymer coating with compositional flexibility as well as extended stability for studying human mesenchymal stem cells (hMSCs). Copolymers contain a majority of poly(ethylene glycol) methyl ether methacrylate (PEGMEMA) as a cytophobic background with poly(et
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