Incorporation of Plant Virus Particles and Polymers as 2D and 3D Scaffolds to Manipulate Cellular Behaviors

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.

1 . A method, comprising:
 growing and cultivating a genetically modified Tobacco mosaic virus coat protein, wherein the genetically modified Tobacco mosaic virus coat protein comprises a cell binding motif to promote cell attachment, outgrowth or differentiation.   
     
     
         2 . The method of  claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from fibronectin. 
     
     
         3 . The method of  claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from vitronectin. 
     
     
         4 . The method of  claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from osteocalcin. 
     
     
         5 . The method of  claim 1 , wherein the cell binding motif comprises an PHSRN (SEQ ID NO 55) sequence derived from fibronectin. 
     
     
         6 . The method of  claim 1 , wherein the cell binding motif comprises an DGEA (SEQ ID NO: 4) peptide sequence derived from collagen. 
     
     
         7 . The method of  claim 1 , wherein the cell binding motif comprises a P15 peptide sequence derived from collagen. 
     
     
         8 . The method of  claim 1 , wherein the cell attachment, outgrowth and differentiation is further accelerated by using serum-free environment supplemented with growth factors. 
     
     
         9 . The method of  claim 8 , wherein the growth factors comprise a fibroblast growth factor, a transforming growth factor-beta1, or an epidermal growth factor. 
     
     
         10 . The method of  claim 1 , further comprising:
 depositing the modified TMV coat protein onto a substrate such that the coated substrate is configured to be used to promote cell attachment, growth or differentiation.   
     
     
         11 . A method comprising:
 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.   
     
     
         12 . The method of  claim 11 , 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). 
     
     
         13 . The method of  claim 11 , wherein the cell is attached to the mutated virus particle in a serum-free environment. 
     
     
         14 . The method of  claim 11 , further comprising:
 prior to attaching the cell, applying the modified-TMV particle onto a surface of a substrate.   
     
     
         15 . A method of forming a hydrogel, the method comprising:
 incorporating a plurality of Tobacco Mosaic Virus particles into a hydrogel precursor formulation; and   polymerizing the hydrogel precursor formulation to for a hydrogel matrix with the Tobacco Mosaic Virus particles incorporated therein.   
     
     
         16 . The method of  claim 15 , wherein the hydrogel precursor formulation comprises a mixture of alginate in bicarbonate solution and a mild acid. 
     
     
         17 . The method of  claim 15 , further comprising:
 crosslinking the hydrogel matrix.   
     
     
         18 . The method of  claim 15 , wherein the Tobacco Mosaic Virus particles comprise modified-TMV particles. 
     
     
         19 . The method of  claim 15 , further comprising:
 prior to incorporation into the hydrogel precursor formulation, attaching a cell binding motif to a carboxy end of a coat protein of a Tobacco Mosaic Virus particles to form the modified-TMV particles.