Array-based microenvironment for cell culturing, cell monitoring and drug-target validation
First Claim
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1. An apparatus, comprising:
- one or more microchambers;
wherein the one or more of the microchambers are adapted to contain cells;
one or more fluid conduits in fluid communication with the microchamber;
one or more valves adapted to regulate the flow of a fluid to the one or more microchambers;
one or more reagent reservoirs in fluid communication with the one or more fluid conduits, wherein the valve regulates the flow of fluids between the one or more microchambers and the one or more reagent reservoirs adapted to exchange nutrients, oxygen and remove excretions; and
the one or more microchambers adapted to contain cells is substantially planar.
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Abstract
An apparatus and method for cell culture in microchambers that are formed on or adjacent to a substrate. The microchambers are connected by one or more fluid conduits, valves and reagent reservoirs, is disclosed.
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Citations
41 Claims
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1. An apparatus, comprising:
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one or more microchambers;
wherein the one or more of the microchambers are adapted to contain cells;
one or more fluid conduits in fluid communication with the microchamber;
one or more valves adapted to regulate the flow of a fluid to the one or more microchambers;
one or more reagent reservoirs in fluid communication with the one or more fluid conduits, wherein the valve regulates the flow of fluids between the one or more microchambers and the one or more reagent reservoirs adapted to exchange nutrients, oxygen and remove excretions; and
the one or more microchambers adapted to contain cells is substantially planar. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37)
the fluid conduit;
the microchamber; and
the reagent reservoir.
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8. The apparatus of claim 1, further comprising an array of arrays coupled to the substrate.
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9. The apparatus of claim 1, further comprising an array associated with the one or more microchambers.
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10. The apparatus of claim 1, wherein the one or more microchambers form a part of an array.
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11. The apparatus of claim 1, further comprising encapsulation of the one or more microchambers, one or more fluid conduits and one or more fluid reservoirs.
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12. The apparatus of claim 1, wherein the one or more microchambers comprise a cell adherent surface.
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13. The apparatus of claim 1, wherein the microchambers form a 4-, 6-, 12-, 48-, 96- or 384-microchamber array.
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14. The apparatus of claim 1, further comprising a diode array in fluid communication with the microchamber or the reagent reservoir.
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15. The apparatus of claim 1, further comprising a micromirror array adjacent to and facing the microchamber or the reagent reservoir.
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16. The apparatus of claim 15, wherein the microchamber further comprises a chemically treated substrate integral with the microchamber suitable for culturing cells wherein the micromirror array directs a light source onto the chemically treated substrate to produce a patterned array of photochemically treated substrate in the microchamber.
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17. The apparatus of claim 16, wherein the light source is integral to the apparatus.
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18. The apparatus of claim 16, wherein the light source comprises a laser.
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19. The apparatus of claim 16, wherein the light source comprises ultraviolet light.
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20. The apparatus of claim 16, wherein the substrate is treated with an organosilane.
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21. The apparatus of claim 20, wherein the organosilane further comprises a biologically functional group.
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22. The apparatus of claim 21, wherein the biologically functional group comprises a free amino moiety.
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23. The apparatus of claim 22, wherein lectin is reacted with the free amino moiety to provide a cell adherent substrate.
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24. The apparatus of claim 16, wherein a microprocessor controls the micromirror direction of the light source onto the substrate.
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25. The apparatus of claim 16, wherein the micromirrors are positioned to direct light into the microchamber to regulate the temperature of the microchamber.
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26. The apparatus of claim 16, wherein the patterned array is produced in a covered microwell.
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27. A method for producing a patterned.array of cultured cells in the cell culture apparatus of claim 16, the method comprising the steps of:
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treating the substrate with one or more photosensitive chemicals;
directing light of a selected wavelength at the micromirrors whereby the micromirrors re-direct the light onto the treated substrate in an array pattern to produce a substrate having photochemically treated portions wherein the photochemically treated portions of the substrate form a patterned array of photochemically treated substrate; and
adhering cells to the substrate whereby the cells adhere to the photochemically treated portions of the substrate to produce a patterned array of cultured cells.
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28. The method of claim 27, wherein the light source comprises a laser.
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29. The method of claim 27, wherein the light source comprises ultraviolet light.
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30. The method of claim 27, wherein the substrate is treated with an organosilane.
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31. The method of claim 30, wherein the organosilane further comprises a biologically functionaligroup.
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32. The method of claim 31, wherein the biologically functional group comprises a free amino moiety.
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33. The method of claim 32, wherein lectin is reacted with the free amino moiety to provide a cell adherent substrate.
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34. The method of claim 27, wherein a computer controls the micromirror direction of the light source onto the substrate.
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35. The method of claim 27, further comprising a laser in fluid communication with the microchamber or the reagent reservoir.
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36. The method of claim 27, further comprising a light amplifier in fluid communication with the microchamber or the reagent reservoir.
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37. The method of claim 27, further comprising a mirror in fluid communication with the microchamber or the reagent reservoir.
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38. An apparatus, comprising:
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one or more microchambers;
wherein the one or more of the microchambers are adapted to contain cells;
one or more fluid conduits in fluid communication with the microchamber;
one or more valves adapted to regulate the flow of a fluid to the one or more microchambers;
one or more reagent reservoirs in fluid communication with the one or more fluid conduits, wherein the valve regulates the flow of fluids between the one or more microchambers and the one or more reagent reservoirs adapted to exchange nutrients, oxygen and remove excretions;
the one or more microchambers adapted to contain cells is substantially planar; and
one or more micromirrors arranged to direct light into the microchambers.
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39. A cell culturing system, comprising:
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one or more nano-scale microchambers for housing cultured cells;
one or more of nano-scale fluid inlets and outlets in fluid communication with the microchambers;
one or more controls to regulate the inlet and the outlet of fluids between the inlets and the outlets and the microchambers; and
one or more controls in communication with the microchambers for detecting biological parameters of the cultured cells.
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40. An apparatus, comprising:
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one or more microchambers;
wherein the one or more of the microchambers are adapted to contain cells;
one or more fluid conduits in fluid communication with the microchamber;
one or more valves adapted to regulate the flow of a fluid to the one or more microchambers;
one or more reagent reservoirs in fluid communication with the one or more fluid conduits, wherein the valve regulates the flow of fluids between the one or more microchambers and the one or more reagent reservoirs adapted to exchange nutrients, oxygen and remove excretions; and
the one or more microchambers adapted to contain cells is substantially planar for detecting biological parameters of the cultured cells.
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41. A cell culturing system comprising:
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a chamber having a bottom substrate for growing cultured cells, the chamber further comprising through-holes for controlled porosity;
a nanoporous semipermeable membrane attached to the substrate; and
a top head on the bottom substrate, the head being adapted to provide space for cells to grow on the substrate and to sandwich the membrane between the substrate and the head, wherein fluid exchange occurs across the membrane through the through-holes.
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Specification