CHEMICALLY-DEFINED ARRAYS FOR SCREENING CELL-SUBSTRATE INTERACTIONS
Patterned SAM arrays and methods of preparing patterned SAM arrays are disclosed. Advantageously, the methods used to prepare the patterned SAM arrays allow for controlling SAM spot-to-spot conditions such as ligand identity and ligand density, which allows for preparing a wide range of SAM spots in a single array format. Additionally, the patterned SAM arrays of the present disclosure support the culture of a range of cell types. The patterned SAM arrays offer the ability to rapidly screen substrate components for influencing cell attachment, spreading, proliferation, migration, and differentiation.
1 .- 20 . (canceled)
21 . A method of screening a cell-surface interaction comprising:
preparing a self-assembled monolayer array, wherein the self-assembled monolayer array is prepared by
adhering a polymer stencil to a metal-coated substrate, wherein the polymer stencil comprises at least one well;
forming at least one alkanethiolate self-assembled monolayer spot on the metal-coated substrate, wherein the alkanethiolate self-assembled monolayer spot is formed in the at least one well of the polymer stencil;
removing the polymer stencil from the metal-coated substrate; and
backfilling a region on the metal-coated substrate that surrounds the at least one alkanethiolate self-assembled monolayer spot, wherein the backfilling forms an alkanethiolate self-assembled monolayer surrounding the at least one alkanethiolate self-assembled monolayer spot;
contacting the cell with the self-assembled monolayer array;
culturing the cell; and
analyzing the cell.
22 . The method of claim 21 , wherein the at least one alkanethiolate self-assembled monolayer spot further comprises a ligand.
23 . The method of claim 22 , wherein the ligand is selected from the group consisting of a protein, a nucleic acid, a polysaccharide, a lipid and combinations thereof.
24 . The method of claim 22 , wherein the at least one alkanethiolate self-assembled monolayer spot further comprises a ligand density range up to about 7.7 pmol/mm 2 .
25 . The method of claim 21 , wherein the alkanethiolate self-assembled monolayer that surrounds the at least one alkanethiolate self-assembled monolayer spot further comprises a ligand.
26 . The method of claim 25 , wherein the ligand is selected from the group consisting of a protein, a nucleic acid, a polysaccharide, a lipid and combinations thereof.
27 . The method of claim 21 , wherein the cell is selected from the group consisting of a mesenchymal stem cell, an embryonic stem cell, an induced pluripotent stem cell, an umbilical vein endothelial cell, a NIH 3T3 fibroblast, a fibrosarcoma cell, a dermal fibroblast and combinations thereof.
28 . The method of claim 21 , further comprising contacting the cell with a soluble molecule.
29 . The method of claim 28 , wherein the soluble molecule is selected from the group consisting of a growth factor and a proteoglycan.
30 . The method of claim 29 , wherein the growth factor is selected from the group consisting of a transforming growth factor beta, a fibroblast growth factor, a platelet derived growth factor and combinations thereof.
31 . The method of claim 29 , wherein the proteoglycan comprises a side chain selected from the group consisting of a heparin glycosaminoglycan side chain, a heparan glycosaminoglycan side chain, a chondroitin glycosaminoglycan side chain and combinations thereof.
32 . A method of screening a cell-surface interaction comprising:
preparing a self-assembled monolayer array, wherein the self-assembled monolayer array is prepared by
adhering a polymer stencil to a metal-coated substrate, wherein the polymer stencil comprises at least one well;
forming at least one alkanethiolate self-assembled monolayer spot on the metal-coated substrate, wherein the alkanethiolate self-assembled monolayer spot is formed in the at least one well of the polymer stencil;
removing the polymer stencil from the metal-coated substrate; and
backfilling a region on the metal-coated substrate that surrounds the at least one alkanethiolate self-assembled monolayer spot, wherein the backfilling forms an alkanethiolate self-assembled monolayer surrounding the at least one alkanethiolate self-assembled monolayer spot;
contacting the cell with the at least one alkanethiolate self-assembled monolayer spot;
culturing the cell; and
analyzing the cell.
33 . The method of claim 32 , wherein the cell is selected from the group consisting of a mesenchymal stem cell, an embryonic stem cell, an induced pluripotent stem cell, an umbilical vein endothelial cell, a NIH 3T3 fibroblast, a fibrosarcoma cell, a dermal fibroblast and combinations thereof.
34 . The method of claim 32 , further comprising contacting the cell with a soluble molecule.
35 . The method of claim 34 , wherein the soluble molecule is selected from the group consisting of a growth factor and a proteoglycan.
36 . The method of claim 32 , wherein the at least one alkanethiolate self-assembled monolayer spot further comprises a ligand.
37 . The method of claim 36 , wherein the ligand is selected from the group consisting of a protein, a nucleic acid, a polysaccharide, a lipid and combinations thereof.
38 . The method of claim 36 , wherein the at least one alkanethiolate self-assembled monolayer spot further comprises a ligand density range up to about 7.7 pmol/mm 2 .
39 . The method of claim 32 , wherein the alkanethiolate self-assembled monolayer surrounding the at least one alkanethiolate self-assembled monolayer spot further comprises a ligand.
40 . The method of claim 39 , wherein the ligand is selected from the group consisting of a protein, a nucleic acid, a polysaccharide, a lipid and combinations thereof.