Covalently-immobilized hydrogel arrays in multi-well plates
Hydrogel arrays, methods for preparing hydrogel arrays and methods for screening cell-substrate interactions using the hydrogel arrays are disclosed. Advantageously, the hydrogel arrays include individual hydrogel posts that are completely isolatable, allowing for systematic and independent control of the chemical composition and physical dimensions of each hydrogel post.
1. A method of preparing a hydrogel array comprising a multi-well plate, the multi-well plate comprising a plurality of wells wherein the plurality of wells independently comprises a hydrogel post, the method comprising
thiol-functionalizing a bottom surface of the plurality of wells;
adding a hydrogel precursor solution to at least one well of the plurality of wells;
selectively polymerizing a portion of the hydrogel precursor solution in the at least one well;
removing unpolymerized hydrogel precursor solution from the plurality of wells, wherein the polymerized hydrogel forms a hydrogel post covalently immobilized within the at least one well and wherein the hydrogel post is surrounded by a hydrogel-free background; and
forming an additional hydrogel layer on the hydrogel post.
2. The method of claim 1 wherein the thiol-functionalizing of the bottom surface of the plurality of the wells is by silanization.
3. The method of claim 1 further comprising assembling the multi-well plate with a negative insert, wherein the negative insert fills a portion of a volume of the well.
4. The method of claim 1 wherein the multi-well plate is selected from the group consisting of a glass-bottom multi-well plate and a polystyrene multi-well plate.
5. The method of claim 4 , wherein the multi-well plate is selected from the group consisting of 4-well plates, 6-well plates, 8-well plates, 12-well plates, 24-well plates, 32-well plates, 96-well plates, and 384-well plates.
6. The method of claim 1 wherein the hydrogel precursor solution is polymerized by exposure to ultraviolet light.
7. The method of claim 1 wherein the hydrogel precursor solution comprises a ligand.
8. The method of claim 7 wherein the ligand is selected from consisting of a protein, a peptide, a nucleic acid, a polysaccharide, a lipid, and combinations thereof.
9. The method of claim 1 wherein the at least one hydrogel post comprises a variable modulus, a variable ligand identity, a variable ligand density and combinations thereof.
10. A method of preparing a hydrogel array comprising a multi-well plate, the multi-well plate comprising a plurality of wells wherein at least one well independently comprises multiple hydrogel posts, the method comprising:
thiol-functionalizing a bottom surface of the plurality of wells;
adding a hydrogel precursor solution to at least one well of the plurality of wells;
assembling the multi-well plate with a negative insert for each of the at least one well, wherein the negative insert comprises wells configured to allow multiple hydrogel posts to be formed in the well assembled with the negative insert;
selectively polymerizing a portion of the hydrogel precursor solution in the at least one well; and
removing unpolymerized hydrogel precursor solution from the plurality of wells, wherein the polymerized hydrogel forms multiple hydrogel posts covalently immobilized within each of the at least one well.