# US 2010/0167330 A1 · Micromechanical devices for control of cell-cell interaction, and methods of use thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2010/0167330 A1 - Document ID: US20100167330A1 - Status: Published application - Kind: A1 - Application number: 12/449,994 - Filed: 2008-03-10 - Published: 2010-07-01 - Earliest claimed: 2007-03-08 - Assignees: The Regents of the University of California - Inventors: Sangeeta N. Bhatia; Elliot Hui - Web page: https://patentgrep.com/patent/US20100167330A1 ## CPC - C12M 35/08: {Chemical, biochemical or biological means, e.g. plasma jet, co-culture} - C12M 23/04: {Flat or tray type, drawers (C12M23/10, C12M23/12, C12M23/16 take precedence)} - C12M 23/20: {Material Coatings (immunocoatings C12M25/00)} - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} ## Abstract The development and function of living tissues depends largely on interactions between cells that can vary in both time and space; however, temporal control of cell-cell interaction is experimentally challenging. By employing a micromachined silicon substrate with moving parts, herein is disclosed the dynamic regulation of cell-cell interactions via direct manipulation of adherent cells with micron-scale precision. The inventive devices and methods allow mechanical control of both tissue composition and spatial organization. The inventive device and methods enable the investigation of dynamic cell-cell interaction in a multitude of applications, such as intercellular communication, spanning embryogenesis, homeostasis, and pathogenic processes. ## Claims ### Claim 1 A method, comprising the steps of: culturing at least one cell of a first type on a first component; culturing at least one cell of a second type on a second component; and placing the first component at a distance from or in contact with the second component for a period of time; thereby co-culturing the at least one cell of a first type and the at least one cell of a second type. ### Claim 2 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; the at least one cell of a second type is a plurality of cells of a second type; the cells of a first type are stem cells or progenitor cells; the cells of a second type secrete a differentiation-inducing signal; and the co-culturing results in stimulating cell differentiation. ### Claim 3 (depends on 2) The method of claim 2, further comprising the step of determining if the cells of a first type have differentiated. ### Claim 4 (depends on 2) The method of claim 2, wherein the cells of a first type are mammalian cells. ### Claim 5 (depends on 2) The method of claim 2, wherein the cells of a first type are human cells. ### Claim 6 (depends on 2) The method of claim 2, wherein the cells of a first type are early embryonic stem cells, blastocyst embryonic stem cells, fetal stem cells, umbilical cord stem cells, or adult stem cells. ### Claim 7 (depends on 2) The method of claim 2, wherein the cells of a first type are adult stem cells isolated from nerve cells, blood cells, muscle cells, skin cells, or bone cells. ### Claim 8 (depends on 2) The method of claim 2, wherein the cells of a first type are human embryonic stem cells. ### Claim 9 (depends on 2) The method of claim 2, wherein the cells of a second type are mammalian cells. ### Claim 10 (depends on 2) The method of claim 2, wherein the cells of a second type are human cells. ### Claim 11 (depends on 2) The method of claim 2, wherein the cells of a second type secrete morphogens. ### Claim 12 (depends on 2) The method of claim 2, wherein the cells of a second type secrete morphogens selected from the group consisting of decapentaplegic/transforming growth factor beta, hedgehog/sonic hedgehog, wingless/wnt, epidermal growth factor, and fibroblast growth factor. ### Claim 13 (depends on 2) The method of claim 2, wherein the cells of a second type secrete morphogens and are selected from the group consisting of human OP-1, mouse OP-1, human OP-2, mouse OP-2, 60A, GDF-1, BMP2A, BMP2B, DPP, Vgl, Vgr-1, BMP3, BMP5, and BMP6. ### Claim 14 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; the at least one cell of a second type is a plurality of cells of a second type; the cells of a first type are target cells comprising a receptor; the cells of a second type secrete soluble signal; and the co-culturing results in exposing cells to cytokines. ### Claim 15 (depends on 14) The method of claim 14, further comprising the step of determining the effect of soluble signal exposure on the cells of a first type. ### Claim 16 (depends on 14) The method of claim 14, wherein the cells of a first type are mammalian cells. ### Claim 17 (depends on 14) The method of claim 14, wherein the cells of a first type are human cells. ### Claim 18 (depends on 14) The method of claim 14, wherein the cells of a first type comprise receptors for a member of the VEGF family, VEGF-D, a member of the MIP family, MIP-1γ, ceruloplasmin, nitric oxide, gases, or growth factors. ### Claim 19 (depends on 14) The method of claim 14, wherein the cells of a first type comprise DLK, Dlk-1, a cahedrins, or T-cahedrin. ### Claim 20 (depends on 14) The method of claim 14, wherein the cells of a second type are mammalian cells. ### Claim 21 (depends on 14) The method of claim 14, wherein the cells of a second type are human cells. ### Claim 22 (depends on 14) The method of claim 14, wherein the cells of a second type secrete hematopoietins, interferons, tumor necrosis factors, chemokines, or a combination thereof. ### Claim 23 (depends on 14) The method of claim 14, wherein the cells of a second type secrete a member of the VEGF family, VEGF-D, a member of the MIP family, MIP-1γ, ceruloplasmin, nitric oxide, gases, or growth factors ### Claim 24 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; the at least one cell of a second type is a plurality of cells of a second type; the cells of a first type are target cells comprising a cytokine receptor; the cells of a second type secrete a cytokine; and the co-culturing results in exposing cells to cytokines. ### Claim 25 (depends on 24) The method of claim 24, further comprising the step of determining the effect of cytokine exposure on the cells of a first type. ### Claim 26 (depends on 24) The method of claim 24, wherein the cells of a first type are mammalian cells. ### Claim 27 (depends on 24) The method of claim 24, wherein the cells of a first type are human cells. ### Claim 28 (depends on 24) The method of claim 24, wherein the cells of a first type comprise cytokine receptors selected from the group consisting of hematopoietin family receptors, interferon family receptors, tumor necrosis factor family receptors, and chemokine family receptors. ### Claim 29 (depends on 24) The method of claim 24, wherein the cells of a first type comprise cytokine receptors selected from the group consisting of receptors for IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-12, IL-15, GM-CSF, IFN-α, IFN-β, IFN-γ, TNF-α, TNF-β, CD40, Fas, MIP-1α, MIP-1β, RANTES, CCR5, and CXCR4. ### Claim 30 (depends on 24) The method of claim 24, wherein the cells of a second type are mammalian cells. ### Claim 31 (depends on 24) The method of claim 24, wherein the cells of a second type are human cells. ### Claim 32 (depends on 24) The method of claim 24, wherein the cells of a second type secrete hematopoietins, interferons, tumor necrosis factors, chemokines, or a combination thereof. ### Claim 33 (depends on 24) The method of claim 24, wherein the cells of a second type secrete IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-12, IL-15, GM-CSF, IFN-α, IFN-β, IFN-γ, TNF-α, TNF-β, CD40, Fas, MIP-1α, MIP-1β, RANTES, CCR5, or CXCR4. ### Claim 34 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; the at least one cell of a second type is a plurality of cells of a second type; the cells of a first type are tumor cells; the cells of a second type are not tumor cells; and the co-culturing results in exposing the tumor cells to the non-tumor cells. ### Claim 35 (depends on 34) The method of claim 34, further comprising the step of determining changes in biology of the cells of a first type. ### Claim 36 (depends on 34) The method of claim 34, wherein the cells of a first type are mammalian cells. ### Claim 37 (depends on 34) The method of claim 34, wherein the cells of a first type are human cells. ### Claim 38 (depends on 34) The method of claim 34, wherein the cells of a first type are selected from the group consisting of tumor cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 39 (depends on 34) The method of claim 34, wherein the cells of a first type are tumor cells selected from the group consisting of carcinoma, lymphomas, leukemias, sarcomas, mesotheliomas, gliomas, germinomas, and choriocarcinomas. ### Claim 40 (depends on 34) The method of claim 34, wherein the cells of a second type are mammalian cells. ### Claim 41 (depends on 34) The method of claim 34, wherein the cells of a second type are human cells. ### Claim 42 (depends on 34) The method of claim 34, wherein the cells of a second type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 43 (depends on 34) The method of claim 34, wherein the cells of a second type are cancer-associated cells. ### Claim 44 (depends on 34) The method of claim 34, wherein the cells of a second type are fibroblasts. ### Claim 45 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; the at least one cell of a second type is a plurality of cells of a second type; and the cells of a second type secrete the soluble factor; further comprising the step of isolating the soluble factor. ### Claim 46 (depends on 45) The method of claim 45, further comprising the step of using RNAi techniques to knockdown expression of the soluble factor. ### Claim 47 (depends on 45) The method of claim 45, further comprising the step of determining the effect of the soluble factor on the cells of a first type. ### Claim 48 (depends on 45) The method of claim 45, wherein the soluble factor has a protective or regenerative effect on the cells of a first type. ### Claim 49 (depends on 45) The method of claim 45, wherein the soluble factor is selected from the group consisting of BDNF, CNTF, Dvl-1, EGF, Endostatin, FGF, GDNF, GM-CSF, Heregulin, IGF, IL, Insulin, Interferon, Jagged1, M-CSF, NAG-1, NGF, NT, PDGF, PEDF, Prolactin, SDF, SF-1, TGF, TNF, VEGF, and Wnt. ### Claim 50 (depends on 45) The method of claim 45, wherein the soluble factor is selected from the group consisting of fibrinogen, laminin, collagen N, tenascin, fibronectin, collagen, bovine pituitary extract, EGF, hepatocyte growth factor, keratinocyte growth factor, hydrocortisone, dimethyl sulphoxide, recombinant human epidermal growth factor, insulin, sodium selenite, transferrin, hydrocortisone, basic fibroblast growth factor, and leukemia inhibitory factor. ### Claim 51 (depends on 45) The method of claim 45, wherein the cells of a first type are mammalian cells. ### Claim 52 (depends on 45) The method of claim 45, wherein the cells of a first type are human cells. ### Claim 53 (depends on 45) The method of claim 45, wherein the cells of a first type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 54 (depends on 45) The method of claim 45, wherein the cells of a second type are mammalian cells. ### Claim 55 (depends on 45) The method of claim 45, wherein the cells of a second type are human cells. ### Claim 56 (depends on 45) The method of claim 45, wherein the cells of a second type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 57 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; and the at least one cell of a second type is a plurality of cells of a second type; further comprising the steps of separating the first component from the second component; exposing the cells of a first type on the first component to a compound for a time; and placing the first component in proximity to or in contact with the second component; thereby selectively exposing a subpopulation of cells from a mixed culture of cells to a compound. ### Claim 58 (depends on 57) The method of claim 57, wherein the compound is a toxic compound, exposure to which results in the death of at least some of the cells of a first type. ### Claim 59 (depends on 57) The method of claim 57, wherein the cells of a first type are mammalian cells. ### Claim 60 (depends on 57) The method of claim 57, wherein the cells of a first type are human cells. ### Claim 61 (depends on 57) The method of claim 57, wherein the cells of a first type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 62 (depends on 57) The method of claim 57, wherein the cells of a second type are mammalian cells. ### Claim 63 (depends on 57) The method of claim 57, wherein the cells of a second type are human cells. ### Claim 64 (depends on 57) The method of claim 57, wherein the cells of a second type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 65 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; and the at least one cell of a second type is a plurality of cells of a second type; further comprising the step of exposing the cells of a first type on the first component and the cells of a second type on a second component to a compound for a time, thereby selectively exposing a mixed culture of cells to a compound. ### Claim 66 (depends on 65) The method of claim 65, wherein the compound is a toxic compound, exposure to which results in the death of at least some of the cells of a first type, some of the cells of a second type, or cells of both types. ### Claim 67 (depends on 65) The method of claim 65, wherein the cells of a first type are mammalian cells. ### Claim 68 (depends on 65) The method of claim 65, wherein the cells of a first type are human cells. ### Claim 69 (depends on 65) The method of claim 65, wherein the cells of a first type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 70 (depends on 65) The method of claim 65, wherein the cells of a second type are mammalian cells. ### Claim 71 (depends on 65) The method of claim 65, wherein the cells of a second type are human cells. ### Claim 72 (depends on 65) The method of claim 65, wherein the cells of a second type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 73 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is a plurality of cells of a first type; and the at least one cell of a second type is a plurality of cells of a second type; further comprising the steps of separating the first component from the second component; and assaying the cells of a first type, the cells of the second type, or both; thereby selectively assaying a subpopulation of cells from a mixed culture of cells. ### Claim 74 (depends on 73) The method of claim 73, wherein the assay is a viability assay. ### Claim 75 (depends on 73) The method of claim 73, wherein the cells of a first type are mammalian cells. ### Claim 76 (depends on 73) The method of claim 73, wherein the cells of a first type are human cells. ### Claim 77 (depends on 73) The method of claim 73, wherein the cells of a first type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 78 (depends on 73) The method of claim 73, wherein the cells of a second type are mammalian cells. ### Claim 79 (depends on 73) The method of claim 73, wherein the cells of a second type are human cells. ### Claim 80 (depends on 73) The method of claim 73, wherein the cells of a second type are selected from the group consisting of cells of the anus, bladder, bile duct, bone, brain, breast, cervix, colon/rectum, duodenum, endometrium, esophagus, eye, gallbladder, head, neck, liver, larynx, lung, mouth, pancreas, penis, prostate, kidney, ovaries, skin, stomach, testicles, and thyroid. ### Claim 81 (depends on 1) The method claim 1, wherein the distance is in the range of about 1 μm to about 1,000 μm. ### Claim 82 (depends on 1) The method of claim 1, wherein the distance is in the range of about 10 μm to about 200 μm. ### Claim 83 (depends on 1) The method of claim 1, wherein the distance is in the range of about 50 μm to about 100 μm. ### Claim 84 (depends on 1) The method of claim 1, wherein the first component comprises a first plurality of fingers. ### Claim 85 (depends on 84) The method of claim 84, wherein each finger in the first plurality of fingers is tapered. ### Claim 86 (depends on 84) The method of claim 84, wherein the ratio of the lengths of the ends of each finger in the first plurality of fingers is about 3:1. ### Claim 87 (depends on 1) The method of claim 1, wherein the second component comprises second a plurality of fingers. ### Claim 88 (depends on 87) The method of claim 87, wherein each finger in the second plurality of fingers is tapered. ### Claim 89 (depends on 87) The method of claim 87, wherein the ratio of the lengths of the ends of each finger in the second plurality of fingers is about 3:1. ### Claim 90 (depends on 87) The method of claim 87, wherein the length of each finger in the first plurality of fingers is about 1 mm to about 50 mm. ### Claim 91 (depends on 87) The method of claim 87, wherein the length of each finger in the first plurality of fingers is about 1 mm to about 1 mm. ### Claim 92 (depends on 87) The method of claim 87, wherein the length of each finger in the first plurality of fingers is about 5 mm. ### Claim 93 (depends on 87) The method of claim 87, wherein the length of each finger in the second plurality of fingers is about 1 mm to 50 mm. ### Claim 94 (depends on 87) The method of claim 87, wherein the length of each finger in the second plurality of fingers is about 1 mm to about 10 mm. ### Claim 95 (depends on 87) The method of claim 87, wherein the length of each finger in the second plurality of fingers is about 5 mm. ### Claim 96 (depends on 1) The method of claim 1, wherein the first component comprises at least or exactly two latches, and the latches are on opposite sides of the first component. ### Claim 97 (depends on 1) The method of claim 1, wherein the second component comprises at least or exactly two latches, and the latches are on opposite sides of the second component. ### Claim 98 (depends on 1) The method of claim 1, wherein the first component comprises at least or exactly two slots, and the slots are on opposite sides of the first component. ### Claim 99 (depends on 1) The method of claim 1, wherein the first component comprises at least or exactly four slots, and the slots are on opposite sides of the first component. ### Claim 100 (depends on 1) The method of claim 1, wherein the first component comprises at least or exactly six slots, and the slots are on opposite sides of the first component. ### Claim 101 (depends on 1) The method of claim 1, wherein the second component comprises at least or exactly two slots, and the slots are on opposite sides of the second component. ### Claim 102 (depends on 1) The method of claim 1, wherein the second component comprises at least or exactly four slots, and the slots are on opposite sides of the second component. ### Claim 103 (depends on 1) The method of claim 1, wherein the second component comprises at least or exactly six slots, and the slots are on opposite sides of the second component. ### Claim 104 (depends on 1) The method of claim 1, wherein the slots are V-shaped. ### Claim 105 (depends on 1) The method of claim 1, wherein the first component is fabricated from silicon, polystyrene, quartz, glass, fused silica, SU-8, PDMS, polypropylene, epoxies, polymers, ceramics or metals. ### Claim 106 (depends on 1) The method of claim 1, wherein the second component is fabricated from silicon, polystyrene, quartz, glass, fused silica, SU-8, PDMS, polypropylene, epoxies, polymers, ceramics or metals. ### Claim 107 (depends on 1) The method of claim 1, wherein the first component is partially or completely coated with polystyrene. ### Claim 108 (depends on 1) The method of claim 1, wherein the second component is partially or completely coated with polystyrene. ### Claim 109 (depends on 1) The method of claim 1, wherein the first component is partially or completely coated with collagen. ### Claim 110 (depends on 1) The method of claim 1, wherein the second component is partially or completely coated with collagen. ### Claim 111 (depends on 1) The method of claim 1, wherein the first component is fabricated from an optically transparent material. ### Claim 112 (depends on 1) The method of claim 1, wherein the first component is fabricated from an optically translucent material. ### Claim 113 (depends on 1) The method of claim 1, wherein the second component is fabricated from an optically transparent material. ### Claim 114 (depends on 1) The method of claim 1, wherein the second component is fabricated from an optically translucent material. ### Claim 115 (depends on 1) The method of claim 1, wherein the at least one cell of a first type is not the same as the at least one cell of the second type. ### Claim 116 (depends on 1) The method of claim 1, further comprising the step of altering the distance between the first component and the second component. ### Claim 117 (depends on 1) The method of claim 1, further comprising the step of replacing the first component or the second component with a third component; wherein the third component wherein the third component comprises at least one cell of a third type, different from the first type of cell and the second type of cell.