Three-dimensional cellular arrays, methods of making three-dimensional cellular arrays, and methods of identifying agents using the arrays are disclosed.
대표청구항▼
1. A cellular system comprising: a porous, hydrophilic substrate that wicks fluids by capillary action selected from the group consisting of paper, nitrocellulose, cellulose acetate, cloth, and porous polymer film, wherein the substrate comprises at least one porous region, each porous region bounde
1. A cellular system comprising: a porous, hydrophilic substrate that wicks fluids by capillary action selected from the group consisting of paper, nitrocellulose, cellulose acetate, cloth, and porous polymer film, wherein the substrate comprises at least one porous region, each porous region bounded at least in part by a liquid impervious boundary, and wherein at least one of the porous regions comprises a temperature-sensitive hydrogel comprising cells. 2. The cellular system of claim 1, wherein the liquid impervious boundary is selected from the group consisting of poly(dimethylsiloxane), poly(lactic-co-glycolic acid), epoxy, polystyrene, a polyether, a polyamide, poly(methyl methacrylate), polycarbonate, polyethylene, polypropylene, a photoresist precursors, a wax and a fat and mixtures thereof. 3. The cellular system of claim 1, further comprising: a bottomless microtiter plate having a plurality of wells disposed over the porous hydrophilic substrate;wherein the wells of the microtiter plate and the liquid impervious boundaries of the porous hydrophilic substrate are positioned so that the plurality of wells are aligned and sealingly joined to the plurality of liquid impervious boundaries to form an individual chamber for each porous region, wherein the wells and porous regions define an array. 4. A kit comprising: one or more porous, hydrophilic substrates selected from the group consisting of paper, nitrocellulose, cellulose acetate, cloth, and porous polymer film, wherein the substrate comprises at least one porous region, each porous region bounded at least in part by a liquid impervious boundary; anda bottomless microtiter plate having a plurality of wells equal to the number of porous regions in the substrate, wherein the wells of the microtiter plate and the liquid impervious boundaries of the porous hydrophilic substrate are positioned so that the plurality of wells are aligned. 5. The kit of claim 4, further comprising a temperature-sensitive hydrogel. 6. A method of making a cellular system, comprising: providing a porous, hydrophilic substrate selected from the group consisting of paper, nitrocellulose, cellulose acetate, cloth, and porous polymer film, wherein the porous, hydrophilic substrate comprises at least one porous region, each porous region bounded at least in part by a liquid impervious boundary; andin any order,contacting a plurality of defined regions of the substrate with a suspension comprising cells and a temperature-sensitive hydrogel precursor, wherein the cells and the hydrogel precursor saturate the plurality of defined regions of the substrate; andcontacting the hydrogel precursor with a gelling agent, wherein the gelling agent induces the formation of a hydrogel embedded in the plurality of defined regions of the substrate. 7. The method of claim 6, wherein cells are introduced into the hydrogel precursor before contacting the hydrogel precursor to the substrate. 8. The method of claim 6, wherein the gelling agent is temperature, a solution containing salt, or a chemical cross-linking agent. 9. The method of claim 6, wherein the liquid impervious boundary is one or more selected from the group consisting of poly(dimethylsiloxane), poly(lactic-co-glycolic acid), epoxy, polystyrene, a polyether, a polyamide, poly(methyl methacrylate), polycarbonate, polyethylene, polypropylene, a photoresist precursors, a wax and a fat and mixtures thereof. 10. The method of claim 6, wherein the cells are one or more cells selected from the group consisting of bacterial cells, insect cells, yeast cells, and mammalian cells. 11. The method of claim 6, further comprising contacting the system with living tissue. 12. A method of identifying an agent that modifies cellular activity, the method comprising: contacting the cellular system of claim 1 with one or more test agents; anddetecting one or more cellular activities in the presence of the one or more test agents. 13. The method of claim 12, wherein the cellular system is contacted with the one or more test agents at a plurality of defined regions. 14. The method of claim 13, wherein each defined region is contacted with a different test agent. 15. The method of claim 12, wherein the cellular activity is one or more selected from the group consisting of proliferation, migration, apotosis apoptosis, differentiation, viability, upregulation of gene transcription, or downregulation of gene transcription. 16. The method of claim 12, wherein the test agent is selected from the group consisting of one or more of a small molecule, amino acid, polypeptide, nucleic acid, carbohydrate, polysaccharide, and metabolite. 17. The method of claim 12, wherein the cells are selected from the group consisting of bacterial cells, insect cells, yeast cells, and mammalian cells and mixtures thereof. 18. A method of claim 12, further comprising: cutting the cellular system into a plurality of segments, each segment comprising at least one porous region comprising cells and hydrogel;contacting each segment with a test agent or a control; anddetecting one or more cellular activities in the presence of the test agent. 19. The cellular system of claim 1, wherein the cells are one or more cells selected from the group consisting of bacterial cells, insect cells, yeast cells, and mammalian cells and mixtures thereof. 20. The cellular system of claim 1, wherein there are a plurality of porous regions positioned in the porous hydrophilic substrate in the form of an array.
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