Two distinct mechanisms for the interaction of cells with fibronectin substrata

Abstract

Fibronectin (Fn) was adsorbed onto neutral, sulfonated, imine-conjugated or gelatin coated polystyrene latex beads. In all cases, the Fn coated beads bound effectively to Chinese hamster ovary (CHO) cells in suspension. However, the binding of Fn coated neutral or positively charged imine conjugated bead was inhibited by low concentrations of heparin or heparan sulfate or by treatment of the cells with Flavobacterium heparanase. By contrast, binding of Fn coated sulfonated or gelatin beads was insensitive to inhibition by heparin and to heparanase treatment of cells. Adhesion of CHO cells to Fn coated tissue culture plastic was not sensitive to heparin, whereas adhesion of CHO cells to Fn-coated imine-conjugated plastic was sensitive to heparin. These observations imply that the functional status of Fn can be modulated by the nature of the surface to which the Fn is adsorbed. They further imply that, under some circumstances, the heparin/heparan sulfate binding domains of Fn can play a role in the attachment of Fn to the cell membrane via membrane proteoglycans. Under other circumstances, the interaction of Fn with the cell may primarily involve other receptors for Fn, presumably cell surface glycoproteins.

Adhesion of glycosaminoglycan-deficient chinese hamster ovary cell mutants to fibronectin substrata.

“…(e) Digestion of cells with heparitinase may lead to impaired attachment to and spreading of cells on fibronectin substrata (Gill et al, 1986). (f) Addition of glycosaminoglycans or proteoglycans may interfere with the substrate attachment of cells (Brennan et al, 1983;Schwarz and Juliano, 1985;Rosenberg et al, 1986). These reports all indicate that cell-surface proteoglycans are of importance for cell-substrate adhesion, however they contain only circumstantial evidence and each approach has some weaknesses.…”

Stable heparin-producing cell lines derived from the Furth murine mastocytoma.

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ABSTRACT (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) ,uCi/ml; 40 Ci/mmol) (both from New England Nuclear).Radioactive GAGs were isolated by anion-exchange chromatography and ethanol precipitation as described by Bame and Esko (20), except that samples bound to DEAE-Sephacel (Pharmacia LKB) were washed with 0.25 M NaCl in 20 mM sodium acetate (pH 6.0) and eluted with 2.0 M NaCl in acetate buffer. Samples were 3-eliminated with 0.5 M NaOH for 24 h at 40C and reduced with 1 M NaBH4 (21) to remove any covalently linked peptides.

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“…Intravenously administered heparin inhibits blood coagulation through activation of antithrombin III (6) and depresses arterial smooth muscle cell proliferation (7). When added to cultured cells, heparin activates basic fibroblast growth factor (8,9) and alters cell adhesion to extracellular matrices composed of type V collagen (10) and fibronectin (11). Released heparin from mast cells can induce uptake of low density lipoproteins by macrophages (12).…”

“…Unlike other cell culture models, the mastocytoma lines divide rapidly, produce heparin continuously, and permit nutritional and genetic experiments under defined culture conditions. A preliminary account of this work has been presented (19 (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) ,uCi/ml; 40 Ci/mmol) (both from New England Nuclear).…”

Plasma membrane potential of murine erythroleukemia cells: Approach to measurement and evidence for cell‐density dependence

“…Such a complex picture is not inconceivable if one recalls that at least for one important serum-derived component of extracellular matrix, namely fibronectin, it has been shown that the capacity of this substance to bind to cell membranes is modulated by the physical properties of the culture surface (Schwarz and Juliano, 1985).…”