Chemical modification of promote animal cell adhesion on surfaces

• US 5,278,063 A
• Filed: 09/28/1989
• Issued: 01/11/1994
• Est. Priority Date: 09/28/1989
• Status: Expired due to Term

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Massia, Stephen P., Hubbell, Jeffrey A.
• Primary Examiner(s): Elliott, George C.
• Application Number:
  US07/414,144
• Time in Patent Office: 1,566 Days
• Field of Search: 435/240.23, 435/240.243, 530/329, 530/330
• US Class Current: 435/402
• CPC Class Codes: A61L 17/145 Coating
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 29/048 Other specific proteins or polypeptides not covered by A61L29/045 - A61L29/047
• CPC Class Codes: A61L 29/085 Macromolecular materials
• CPC Class Codes: A61L 31/047 Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
• CPC Class Codes: A61L 31/10 Macromolecular materials
• CPC Class Codes: C07K 17/08 the carrier being a synthetic polymer
• CPC Class Codes: C08L 89/00 Compositions of proteins; Compositions of derivatives thereof foodstuff preparations A23J3/00
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

A cell culture substrate which promotes cell adhesion and the formation of focal contacts is disclosed. Polymer, ceramic and glass surfaces may be used. The surfaces are derivatized with peptides having less than 12 amino acid residues and containing one of the following sequences of amino acids: GRGD, GYIGSR, GREDV. The peptides are covalently attached to the surface throu

The U.S. Government may have rights in the present invention as relevant developmental work was supported by National Science Foundation Grant CBT-881020268, and National Institutes of Health Grant HL-39714.

Claims

1. A cell culture substrate of less than about 45% water comprising a surface with the terminal amine of a peptide covalently linked thereto, wherein said peptide is at least one of

View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)

2. The cell culture substrate of claim 1, wherein said peptide comprises 4 to 6 amino acid residues.

3. The cell culture substrate of claim 1 wherein the peptide is GRGD, GRGDY or GYIGSRY.

4. The cell culture substrate of claim 1 wherein the peptide is GRGD, GRGDY, or GYIGSRY and is covalently attached to said cell culture substrate at the N-terminal glycine of the peptide.

5. The cell culture substrate of claim 1, wherein said peptide is GRGD, GRGDF, GYIGSR, GYIGSRY GREDV or GREDVY.

6. The cell culture substrate of claim 1 or 5, wherein the peptide is GRGD.

7. The cell culture substrate of claim 1 or 5 wherein the peptide is GYIGSR.

8. The cell culture substrate of claim 1, wherein the peptide is GRGDY or GYIGSRY peptide.

9. The cell culture substrate of claim 1 wherein the peptide is attached to the substrate at a surface concentration of between about 0.5 picomoles/cm² to about 50 picomoles/cm².

10. The cell culture substrate of claim 1, wherein the peptide is attached to the substrate at a surface concentration of between about 0.5 picomoles/cm² to about 20 picomoles/cm².

11. The cell culture substrate of claim 1, wherein the peptide is attached to the substrate at a surface concentration of about 12 picomoles/cm².

12. The cell culture substrate of claim 1, wherein the substrate comprises a material selected from the group consisting of:

a ceramic;

a polymer; and

a metal.

13. The cell culture substrate of claim 12 wherein the metal is titanium.

14. The cell culture substrate of claim 1, wherein the substrate comprises:

a polymer;

or

a ceramic.

15. The cell culture substrate of claim 1 or 5 wherein the substrate comprises a ceramic material.

16. The cell culture substrate of claim 15, wherein the ceramic is glass.

17. The cell culture substrate of claim 16, wherein the glass comprises glycerol propylsilane-bonded glass.

18. The cell culture substrate of claim 16, wherein the polymer is poly(hydroxyethyl methacrylate).

19. The cell culture substrate of claim 1, wherein the substrate comprises a polymer selected from the group consisting of:

a plastic;

a poly (hydroxyethyl methylacrylate);

a poly (ethylene terephthalate);

a poly (tetrafluoroethylene);

a fluorinated ethylene; and

a poly (dimethyl siloxane).

20. The cell culture substrate of claim 19, wherein the polymer is poly(ethylene terephthalate).

21. The cell culture substrate of claim 20, wherein the peptide is GRGD or GYIGSR.

22. The cell culture substrate of claim 1 wherein the substrate is essentially free of peptide throughout the substrate bulk.

23. The cell culture substrate of claim 22, wherein the peptide is within 10 nanometers of the substrate surface.

24. A cell culture substrate capable of supporting fibroblast focal adhesion formation and spreading comprising a surface derivatized with a peptide consisting essentially of GYIGSR, wherein said peptide is covalently linked by the terminal glycine to the surface.

25. A cell culture substrate capable of supporting cell focal adhesion formation and cell spreading comprising a surface derivatized with a peptide selected from the group consisting of GRGD, GRGDF, GRDGY, GYIGSR and GYIGSRY, and wherein the terminal glycine of said peptide is covalently linked to surface hydroxyl groups of the surface.