Compositions, methods, and systems for bead formation using improved polymers
The present disclosure provides systems and methods for making a hydrogel comprising a cell, cell nucleus, or one or more components derived from a cell or cell nucleus. A method for making a hydrogel may comprise providing a cell or cell nucleus, a first polymer, wherein the first polymer comprises a plurality of first crosslink precursors, each of the plurality of first crosslink precursors comprising an azide group; providing a second polymer, wherein the second polymer comprises a plurality of second crosslink precursors, each of the plurality of second crosslink precursors comprising an alkyne group; and crosslinking the first polymer and the second polymer via a reaction between a first section of the first crosslink precursors and a second section of the second crosslink precursors, thereby providing the hydrogel comprising the cell or cell nucleus.
1. A method comprising:
(a) providing a live cell encapsulated in a gel matrix that is diffusively permeable to chemical or biochemical reagents, wherein the gel matrix comprises two or more crosslinked polymers, wherein at least one polymer of said two or more crosslinked polymers is selected from the group consisting of a polyolefin, an olefin copolymer, an acrylic, a vinyl polymer, a polyamide, a polyimide, a formaldehyde resin, a polyurethane, a cellulosic, a thermoplastic elastomer, and a thermoplastic polyurethane, and wherein the crosslinks are formed by click chemistry;
(b) culturing the live cell.
2. The method of claim 1 , wherein the method further comprises contacting the gel matrix with a chemical or biochemical reagent.
3. The method of claim 1 , wherein the culturing of the live cell is in the presence of a chemical or a biochemical reagent.
4. The method of claim 1 , wherein the chemical or biochemical reagents are selected from nucleic acids, enzymes, potential cell binding ligands.
5. The method of claim 1 , wherein the gel matrix further comprises an activatable functional group.
6. The method of claim 1 , wherein the method further comprises analyzing one or more components of the live cell.
7. The method of claim 1 , wherein the gel matrix is contained in a partition.
8. The method of claim 1 , wherein the live cell is selected from a prokaryotic cells, eukaryotic cells, a bacterial cell, a fungal cell, a plant cell, a mammalian cell, a mycoplasma cell, a normal tissue cell, a tumor cell, a T-cell, and a rare cell.
9. The method of claim 1 , wherein the crosslinks are distributed throughout the gel.
10. The method of claim 1 , wherein the gel matrix is contained in a partition, wherein the partition is a droplet.
11. The method of claim 1 , wherein the crosslinks comprise a picolyl moiety.
12. The method of claim 1 , wherein the click chemistry is copper-catalyzed or copper-free.
13. The method of claim 12 , wherein the click chemistry is copper-free click chemistry selected from: (a) strain-promoted azide/dibenzocyclooctyne-amine (DBCO) click chemistry; (b) inverse electron demand Diels-Alder (IED-DA) tetrazine/trans-cyclooctene (TCO) click chemistry; (c) inverse electron demand Diels-Alder (IED-DA) tetrazine/norbonene click chemistry; (d) Diels-Alder maleimide/furan click-chemistry; (e) Staudinger ligation; and (f) nitrile-oxide/norbonene cycloaddition click chemistry.
14. The method of claim 1 , wherein the crosslinks comprise a 1,2,3-triazole moiety or a dihydropyridazine moiety.
15. The method of claim 1 , wherein the crosslinks comprise a labile bond.
16. The method of claim 15 , wherein the labile bond is selected from a chemically labile bond, a thermally labile bond, an enzymatically labile bond, a photo-labile bond, a disulfide bond, a carbamate bond, a peptide bond, or a combination thereof.
17. A composition comprising
a partition comprising a live cell encapsulated in a gel matrix that is diffusively permeable to chemical or biochemical reagents, wherein the gel matrix comprises two or more crosslinked polymers, wherein the crosslinks are formed by click chemistry and at least one polymer of said two or more crosslinked polymers is selected from the group consisting of a polyolefin, an olefin copolymer, an acrylic, a vinyl polymer, a polyamide, a polyimide, a formaldehyde resin, a polyurethane, a cellulosic, a thermoplastic elastomer, and a thermoplastic polyurethane.
18. The composition of claim 17 , wherein the crosslinks are distributed throughout the gel matrix.
19. A method of forming a live cell encapsulated in a gel matrix comprising:
combining in a partition under click chemistry reaction conditions (i) a live cell, and (ii) two or more polymers configured to form crosslinks by click chemistry, wherein at least one polymer of said two or more crosslinked polymers is selected from the group consisting of a polyolefin, an olefin copolymer, an acrylic, a vinyl polymer, a polyamide, a polyimide, a formaldehyde resin, a polyurethane, a cellulosic, a thermoplastic elastomer, and a thermoplastic polyurethane.