Extracellular matrix coatings

K. E., Fisher, J. M., Paine, M. F., Lown, K. S., Watkins, P. B., Expression of enzymatically active CYP3A4 by Caco-2 cells grown on extracellular matrix-coated permeable supports in the presence of 1-alpha,25-dihydroxyvitamin D3, Mol. Pharmacol. 1997, 53, 741-754. 

Immediately seed microvessel preparation at a density of 5-10 fragments/cm2 into corneal extracellular matrix-coated dishes containing DMEM supplemented with 20 % CS. 

In vitro, the capillary endothelial cells of the pancreas are still attached to the outer rim of whole islets postisolation and, with conditioned medium, may be coaxed to proliferate outward onto extracellular matrix-coated plastic plates for further mechanistic studies and functional analyses. The isolated capillary endothelial cells can be characterized by examining a-1 proteinase inhibitor and nephrin expression via immunocytochemistry or flow cytometry (Zanone et al., 2008). Not only do these endothelial cells play a role in drug candidate-induced effects on insulin secretion and glucose regulation of islets, but they also act as the regulators of leukocyte recruitment into the islets. The pancreatic microvasculature is therefore likely to play a role in the altered physiology of the endocrine and exocrine pancreas if affected by drug candidate exposure. 

Pham, M.T., Reuther, H., Maitz, M.F., 2003. Native extracellular matrix coating on Ti surfaces. J. Biomed. Mater. Res. Part A 66A, 310-316. 

Ouyang, A., Ng, R., and Yang, S.-T. 2007. Long-term culturing of undifferentiated embryonic stem cells in conditioned media and three-dimensional fibrous matrices without extracellular matrix coating. Stem Cells 25(2) 447-54. 

Fig. 2 Smooth muscle cell migration on the three different extracellular matrix-coated substrate. Cells cultured on fibronectin more migrate into the space than those on vitronectin and type I collagen. Scale bars 500 pm...

Extracellular matrix The surfaces of animal cells are covered with a flexible and sticky layer of complex carbohydrates, proteins, and lipids. This complex coating is cell-specific, serves in cell-cell recognition and communication, creates cell adhesion, and provides a protective outer layer. 

Fibronectin is an extracellular matrix protein that mediates a variety of cellular effects. It is important in cell-cell and cell-substratum interactions ( 3.9), mediates reticuloendothelial cell activity and binds both to Clq (the first component of complement) and to bacteria. It also increases the tu-mouricidal activity of macrophages and activates complement receptors, by regulating the binding of C3b-coated particles to neutrophils. It may mediate attachment of Staphylococcus aureus to neutrophils and may also play a role as an adhesion factor, promoting the adhesion of neutrophils to surfaces. Fibronectin mRNA (8.7-8.8 kb) is detected only at low levels in... 

One TIRF study found that some membrane proteins behave just oppositely to AChR they avoid the cell/substrate contact regions/ 1 When endothelial cells are grown on a bare glass surface or are brought into suspension, a specific membrane protein marked with antibodies appears all over the cell surface, as evidenced by epi-illumination and TIRF. However, when the cells are grown on (or returned to) a surface coated with their own extracellular matrix material, the protein disappears from the basal (cell/substrate-contacting) side of the cells. 

Gateau E, Levasseur P, Borgonovi M, Imbert C. (2007) The effect of aminocandin (HMR 3270) on the in vitro adherence of Candida Albicans to polystyrene surfaces coated with extracellular matrix proteins or fibronectin. Clin Microbiol Infec 13 311-315. 

Heparin has been found to bind a large number of proteins (Table 3). The biological activity of heparin and related polysaccharides is usually ascribed to their interaction with heparin-binding proteins. These proteins can be classified into classes including (1) enzymes, (2) protease inhibitors, (3) lipoproteins, (4) growth factors, (5) chemokines, (6) selectins, (7) extracellular matrix proteins, (8) receptor proteins, (9) viral coat proteins, (10) nuclear proteins, and (11) other proteins (1). Many heparin-binding proteins are enzymes and enzyme inhibitors. For example, proteases in the coagulation cascade, such as factors Ha, IXa, Xa, Xlla, and Villa, are heparin-... 

The use of plastic materials for routine cell culture on the laboratory scale was introduced at the end of the 1960s, and some characteristics of glass surface, such as hydrophobicity and negative charge, were maintained in these materials. Polystyrene is the most widely used plastic material for animal cell adhesion at present, because of its surface characteristics, its low cost, and also its transparency. For more demanding cell lines, the surface has to be submitted to a treatment that involves coating with proteins such as poly-lysine, poly-ornithine, or extracellular matrix-derived proteins such as fibronectin, laminin, and collagens. 

Thick nitrocellulose filter membrane with 3-5 pm diameter pores (Neuro Probe, Cabin John, MD). As with polycarbonate filters, the thicker nitrocellulose filters may also be coated with extracellular matrix proteins to facilitate lymphocyte migration (see Subheading 2.L). 

Another critical step is the placement of the filter to the filled wells of the chamber. Caution should be utilized when placing the filter on the filled wells as excessive movement may result in contamination between the wells. Furthermore, in lymphocyte migration assays, matrix-coated filters should be coated at least 2 h prior to performing the assay. The filter should be extensively washed and thoroughly dried prior to the placement of the filter in the chamber. A wet filter may permit leaking between the wells of the chamber. In addition, nonbound extracellular matrix proteins must be removed by extensive washing as residual matrix may result in a loss of adhesion to the filter. 

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