Chemically-defined arrays for screening cell-substrate interactions

Abstract

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.