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AssigneeBoard of Regents, The University of Texas System
InventorsJeffrey A. Hubbell, Stephen P. Massia, Neil P. Desai
Estimating the term…
An absorbed protein-independent cell-adhesive surface is disclosed. The treated surface comprises a chemically derivatized material to which small peptides, having less than 12 amino acid residues and including YIGSR, RGD or REDV amino acid sequence are covalently linked to. The peptides of the present invention include a terminal glycine amino acid. Tresyl chloride activation of surface hydroxyl moieties provides the active surface sites by which a terminal glycine arm of a selected peptide attaches to form covalent bonds between the substrate and peptide. Peptides high with cell adhesive properties are bound in high efficiency. By way of example, surfaces which may be used in conjunction with the present invention include polymer, metal, and ceramic surfaces. The most preferred polymer surfaces include PHEMA and PET polymer surfaces, with the most preferred glass surfaces being glycophase glass. The present methods also include a pretreatment method which provides hydroxyl moieties to surfaces devoid of readily available hydroxyl moieties. The pretreatment, by way of example, comprises immersion of the surface in a mixture of formaldehyde and acetic acid. Methods of preparing the treated surfaces are also included in the present invention. Also included are surface-treated biomedical implant devices and cell culturing devices. The treated surface promotes an enhanced rate and an enhanced amount of cell adhesion to the surface, independent of media serum concentrations or other absorbed proteins. The treated surfaces of the present invention are thermally stable, reusable, peptide efficient (attached to surface only) and resistant to cell proteolysis. The invention further concerns polymeric substrates with a surface having physically interpenetrating water-soluble polymer chains, and methods for production thereof.
A cell culture substrate containing less than about 45% water comprising a surface with covalently linked peptide at a concentration density of between about 0.001 picomoles and about 50 picomoles/cm.sup.2, wherein said peptide is attached through an N-terminal amine to a hydroxyl group at the substrate surface, wherein said peptide is at least one peptide selected from the group consisting of:
A method of derivatizing a substrate surface comprising:
A cell culture modification method comprising:
A method for derivatizing a substrate surface of poly (hydroxyethyl methacrylate) comprising:
A method for enhancing selective endothelial cell adhesion to a surface comprising covalently attaching GREDV, or GREDVY peptide through the N-terminal amine of the peptide to surface hydroxyl groups on a substrate surface, and plating cells on the surface, wherein said surface supports selective endothelial cell focal adhesion formation and cell spreading.
A method for enhancing endothelial cell adhesion and spreading to a surface comprising chemically derivatizing the surface with a peptide consisting essentially of GREDV whereby the peptide is covalently bound to the surface through the N-terminal glycine of the peptide;
A method for enhancing fibroblast adhesion to a surface comprising chemically derivatizing the surface with a peptide consisting essentially of GYIGSR whereby the peptide is covalently bound to the surface through the N-terminal glycine of the peptide;
A cell culture substrate comprising a surface with a covalently linked peptide at a concentration density of less than about 50 picomoles/cm.sup.2, said peptide being attached through an N-terminal glycine to hydroxyl groups at the substrate surface, wherein said peptide is at least one peptide selected from the of peptides group consisting of:
A method of preparing a cell substrate surface comprising:
A method for promoting selective endothelial cell and fibroblast cell adhesion without platelet adhesion to a surface comprising:
A method for promoting selective endothelial cell and fibroblast cell adhesion without platelet adhesion to a surface comprising;
A surface for the selective adhesion of endothelial cells and fibroblast cells without platelet adhesion comprising a surface with a covalently attached GYIGSR peptide, wherein said peptide is attached through the N-terminal amine to a hydroxyl group on the surface and wherein said surface supports the selective adhesion of endothelial cells and fibroblast cells and excludes platelet adhesion.
A surface for the selective adhesion of endothelial cells and fibroblasts without platelet adhesion comprising a surface with a covalently attached peptide, GPDSGRY, wherein said peptide is attached through the N-terminal amine to hydroxyl groups on the surface, and wherein said surface supports the selective adhesion of endothelial cells and fibroblast cells and excludes platelet adhesion.
A method for promoting selective endothelial cell adhesion without fibroblast or platelet adhesion to a surface comprising:
A surface for the selective adhesion of endothelial cells without fibroblast or platelet adhesion comprising a surface with a covalently attached peptide GREDVY, wherein said peptide is attached through the N-terminal amine to a hydroxyl group on the surface, and wherein said surface supports the selective adhesion of endothelial cells and excludes fibroblast and platelet adhesion.
A cell culture modification method comprising:
A method of derivatizing a substrate surface to support cell focal adhesion formation and cell spreading comprising:
Dependencies are inferred from the claim text.