Incorporation of plant virus particles and polymers as 2D and 3D scaffolds to manipulate cellular behaviors

Patent metadata

Field	Value
Publication number	US-10487309-B2
Application number	US-201715424064-A
Country	US
Kind code	B2
Filing date	Feb 3, 2017
Priority date	Oct 24, 2012
Publication date	Nov 26, 2019
Grant date	Nov 26, 2019

Abstract

Methods are generally disclosed for attaching a cell binding motif to a carboxy end of a coat protein of a Tobacco Mosaic Virus particle to form a modified-TMV particle; and attaching a cell to the cell binding motif of the modified-TMV particle. Methods are also disclosed for incorporated virus particles, e.g., TMV virus particles into hydrogels.

First claim

What is claimed:

1. A method of forming a hydrogel, the method comprising:
forming modified Tobacco Mosaic Virus particles, the modification comprising including a cell binding motif on a carboxy end of a coat protein of the Tobacco Mosaic Virus particles;
incorporating a plurality of the modified Tobacco Mosaic Virus particles into a hydrogel precursor formulation; and
polymerizing the hydrogel precursor formulation to form a hydrogel matrix with the modified Tobacco Mosaic Virus particles incorporated therein.

2. The method of claim 1 , wherein the hydrogel precursor formulation comprises a mixture of alginate in bicarbonate solution and a mild acid.

3. The method of claim 1 , further comprising crosslinking the hydrogel matrix.

4. The method of claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from fibronectin.

5. The method of claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from vitronectin.

6. The method of claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from osteocalcin.

7. The method of claim 1 , wherein the cell binding motif comprises a PHSRN (SEQ ID NO: 55) sequence derived from fibronectin.

8. The method of claim 1 , wherein the cell binding motif comprises an DGEA (SEQ ID NO: 4) peptide sequence derived from collagen.

9. The method of claim 1 , wherein the cell binding motif comprises a P15 peptide sequence derived from collagen.

10. The method of claim 1 , wherein the cell binding motif is selected from the group consisting of GRGDSPG (SEQ ID NO: 1), AVTGRGDSPASS (SEQ ID NO: 2), EDRVPHSRNSIT (SEQ ID NO: 3), DGEA (SEQ ID NO: 4), and GTPGPQGIAGQRGVV (SEQ ID NO: 5).

11. The method of claim 1 , further comprising maintaining the hydrogel in a serum-free environment supplemented with growth factors.

12. The method of claim 11 , wherein the growth factors comprise a fibroblast growth factor, a transforming growth factor-beta 1, or an epidermal growth factor.

13. The method of claim 1 , further comprising seeding the hydrogel with cells.

14. The method of claim 13 , wherein upon the seeding, the cells bind to the virus particles.

15. The method of claim 1 , wherein the method is free of covalent modification of the hydrogel.

16. A method of forming a hydrogel, the method comprising:
incorporating a plurality of modified Tobacco Mosaic Virus particles into a hydrogel precursor formulation;
polymerizing the hydrogel precursor formulation to form a hydrogel matrix with the modified Tobacco Mosaic Virus particles incorporated therein; and
maintaining the hydrogel incorporating the modified Tobacco Mosaic Virus particles therein in a serum-free environment supplemented with growth factors.

17. The method of claim 16 , wherein the growth factors comprise a fibroblast growth factor, a transforming growth factor-beta1, or an epidermal growth factor.

18. The method of claim 16 , wherein the modified Tobacco Mosaic Virus particles comprise a cell binding motif on a carboxy end of a coat protein of the modified Tobacco Mosaic Virus particles.

19. A method of forming a hydrogel, the method comprising:
incorporating a plurality of modified Tobacco Mosaic Virus particles into a hydrogel precursor formulation;
polymerizing the hydrogel precursor formulation to form a hydrogel matrix with the modified Tobacco Mosaic Virus particles incorporated therein; and
seeding the hydrogel incorporating the modified Tobacco Mosaic Virus particles therein with cells.

20. The method of claim 19 , wherein upon the seeding, the cells bind to the modified Tobacco Mosaic Virus particles.

21. The method of claim 19 , wherein the modified Tobacco Mosaic Virus particles comprise a cell binding motif on a carboxy end of a coat protein of the Tobacco Mosaic Virus particles.

Assignees

• Wang Qian
• Lee Lim Andrew
• Univ South Carolina

Inventors

• Wang Qian
• Lee Lim Andrew

Classifications

• C12N5/0068 Primary

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N2501/15

  Transforming growth factor beta (TGF-β) · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N2501/11

  Epidermal growth factor [EGF] · CPC title

• C12N2770/00031

  Uses of virus other than therapeutic or vaccine, e.g. disinfectant · CPC title