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