# US 10,500,311 B2 · Cell structure for brain damage treatment, production method thereof, and brain damage treatment agent > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,500,311 B2 - Document ID: US10500311B2 - Status: Granted - Kind: B2 - Application number: 15/380,350 - Filed: 2016-12-15 - Published: 2017-04-06 - Granted: 2019-12-10 - Earliest claimed: 2014-06-16 - Assignees: FUJIFILM Corporation - Applicants: FUJIFILM Corporation - Inventors: Kentaro Nakamura - Web page: https://patentgrep.com/patent/US10500311B2 ## CPC - A61K 35/44: Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells - A61L 27/3878: {Nerve tissue, brain, spinal cord, nerves, dura mater} - A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells - A61L 27/00: Materials for {grafts or} prostheses or for coating {grafts or} prostheses (dental prostheses A61C13/00; shape or structure of prostheses A61F2/00; use of preparations for artificial teeth A61K6/80; artificial kidneys A61M1/14) - A61L 27/222: {Gelatin} - A61L 27/3808: {Endothelial cells} - A61L 27/3834: {Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells} - A61L 27/56: Porous materials, {e.g. foams or sponges} - C07K 14/78: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] - C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)} - C12N 5/0697: {Artificial constructs associating cells of different lineages, e.g. tissue equivalents (blood vessels C12N5/0691)} - A61L 2430/32: for nerve reconstruction - C12N 2535/10: Patterned coating - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract An object of the present invention is to provide a cell structure for brain damage treatment which does not contain glutaraldehyde and in which it is possible to exhibit a sufficient effect of treating brain damage, a production method thereof, and a brain damage treatment agent. According to the present invention, there is provided a cell structure for brain damage treatment which contains biocompatible macromolecular blocks and at least one kind of cell and in which a plurality of the biocompatible macromolecular blocks are disposed in gaps between a plurality of the cells, in which the tap density of the biocompatible macromolecular block is 10 mg/cm3 to 500 mg/cm3 or a value obtained by dividing a square root of a cross-sectional area in a two-dimensional cross-sectional image of the biocompatible macromolecular block by a peripheral length is 0.01 to 0.13. ## Claims ### Claim 1 A cell structure for brain damage treatment which contains biocompatible macromolecular blocks and at least one kind of cell and in which a plurality of the biocompatible macromolecular blocks are disposed in gaps between a plurality of the cells, wherein the tap density of the biocompatible macromolecular block is 10 mg/cm3 to 500 mg/cm3 or a value obtained by dividing a square root of a cross-sectional area in a two-dimensional cross-sectional image of the biocompatible macromolecular block by a peripheral length is 0.01 to 0.13, wherein the biocompatible macromolecular block is a biocompatible macromolecular block produced through a method including a step (a) of freezing a solution of biocompatible macromolecules through freezing treatment in which the highest internal liquid temperature, which is a liquid temperature in a portion having the highest liquid temperature within the solution, becomes lower than or equal to a temperature which is 3° C. lower than a melting point of a solvent in an unfrozen state, and a step (b) of freeze-drying the frozen biocompatible macromolecules which have been obtained in the step (a). ### Claim 2 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the cells include at least mesenchymal stem cells and/or bone marrow cells. ### Claim 3 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the number of cells, to be administered, per administration is 1.0×105 to 1.0×107 pieces/kg body weight. ### Claim 4 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the brain damage includes brain injury, hypoxic-ischemic brain damage, cerebral infarction, and/or cerebral stroke. ### Claim 5 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the size of one of the biocompatible macromolecular blocks is 10 μm to 300 μm. ### Claim 6 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the thickness or the diameter of the cell structure is 400 μm to 3 cm. ### Claim 7 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the cell structure includes 0.0000001 μg to 1 μg of a biocompatible macromolecular block per cell. ### Claim 8 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the biocompatible macromolecular block consists of a recombinant peptide. ### Claim 9 (depends on 8) The cell structure for brain damage treatment according to claim 8, wherein the recombinant peptide is any of a peptide formed of an amino acid sequence described in SEQ ID No: 1, a peptide formed of an amino acid sequence in which one or a plurality of amino acids are deleted, substituted, or added in the amino acid sequence described in SEQ ID No: 1, and has biocompatibility, or a peptide formed of an amino acid sequence having 80% or more sequence identity to the amino acid sequence described in SEQ ID No: 1, and has biocompatibility. ### Claim 10 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein, in the biocompatible macromolecular block, the biocompatible macromolecules are cross-linked using heat, ultraviolet rays, or enzymes. ### Claim 11 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the biocompatible macromolecular block is in a granule form obtained by grinding a porous body of a biocompatible macromolecule. ### Claim 12 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the method of producing further comprises a step (c) of grinding a porous body which has been obtained in the step (b). ### Claim 13 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein, in the step (a), the highest internal liquid temperature, which is a liquid temperature in a portion having the highest liquid temperature within the solution becomes lower than or equal to a temperature which is 7° C. lower than a melting point of a solvent in an unfrozen state. ### Claim 14 (depends on 1) The cell structure for brain damage treatment according to claim 1, wherein the cell structure is a cell structure obtained by mixing the biocompatible macromolecular blocks and the cells and culturing the mixture for 10 hours or longer. ### Claim 15 (depends on 1) A cell structure for brain damage treatment which is obtained by merging a plurality of the cell structures for brain damage treatment according to claim 1. ### Claim 16 (depends on 1) A method for producing the cell structure for brain damage treatment according to claim 1, the method comprising: a step of mixing cells and biocompatible macromolecular blocks of which the tap density is 10 mg/cm3 to 500 mg/cm3 or a value obtained by dividing a square root of a cross-sectional area in a two-dimensional cross-sectional image by a peripheral length is 0.01 to 0.13, and culturing the mixture for 10 hours or longer. ### Claim 17 (depends on 1) A brain damage treatment agent comprising: the cell structure for brain damage treatment according to claim 1. ### Claim 18 A method for treating brain damage, the method comprising: a step of administering a cell structure for brain damage treatment to a patient with brain damage, wherein the cell structure for brain damage treatment contains biocompatible macromolecular blocks and at least one kind of cell and a plurality of the biocompatible macromolecular blocks are disposed in gaps between a plurality of at least one kind of cell, wherein the tap density of the biocompatible macromolecular block is 10 mg/cm3 to 500 mg/cm3 or a value obtained by dividing a square root of a cross-sectional area in a two-dimensional cross-sectional image of the biocompatible macromolecular block by a peripheral length is 0.01 to 0.13, wherein the biocompatible macromolecular block is a biocompatible macromolecular block produced through a method including a step (a) of freezing a solution of biocompatible macromolecules through freezing treatment in which the highest internal liquid temperature, which is a liquid temperature in a portion having the highest liquid temperature within the solution, becomes lower than or equal to a temperature which is 3° C. lower than a melting point of a solvent in an unfrozen state, and a step (b) of freeze-drying the frozen biocompatible macromolecules which have been obtained in the step (a). ### Claim 19 (depends on 18) The method for treating brain damage according to claim 18, wherein the cells include at least mesenchymal stem cells and/or bone marrow cells. ### Claim 20 (depends on 18) The method for treating brain damage according to claim 18, wherein the number of cells, to be administered, per administration is 1.0×105 to 1.0×107 pieces/kg body weight. ### Claim 21 (depends on 18) The method for treating brain damage according to claim 18, wherein the brain damage includes brain injury, hypoxic-ischemic brain damage, cerebral infarction, and/or cerebral stroke. ### Claim 22 (depends on 18) The method for treating brain damage according to claim 18, wherein the size of one of the biocompatible macromolecular blocks is 10 μm to 300 μm. ### Claim 23 (depends on 18) The method for treating brain damage according to claim 18, wherein the thickness or the diameter of the cell structure is 400 μm to 3 cm. ### Claim 24 (depends on 18) The method for treating brain damage according to claim 18, wherein the cell structure includes 0.0000001 μg to 1 μg of a biocompatible macromolecular block per cell. ### Claim 25 (depends on 18) The method for treating brain damage according to claim 18, wherein the biocompatible macromolecular block consists of a recombinant peptide. ### Claim 26 (depends on 25) The method for treating brain damage according to claim 25, wherein the recombinant peptide is any of a peptide formed of an amino acid sequence described in SEQ ID No: 1, a peptide formed of an amino acid sequence in which one or a plurality of amino acids are deleted, substituted, or added in the amino acid sequence described in SEQ ID No: 1, and has biocompatibility, or a peptide formed of an amino acid sequence having 80% or more sequence identity to the amino acid sequence described in SEQ ID No: 1, and has biocompatibility. ### Claim 27 (depends on 18) The method for treating brain damage according to claim 18, wherein, in the biocompatible macromolecular block, the biocompatible macromolecules are cross-linked using heat, ultraviolet rays, or enzymes. ### Claim 28 (depends on 18) The method for treating brain damage according to claim 18, wherein the biocompatible macromolecular block is in a granule form obtained by grinding a porous body of a biocompatible macromolecule. ### Claim 29 (depends on 18) The method for treating brain damage according to claim 18, wherein the method of producing further comprises a step (c) of grinding a porous body which has been obtained in the step (b). ### Claim 30 (depends on 18) The method for treating brain damage according to claim 18, wherein, in the step (a), the highest internal liquid temperature, which is a liquid temperature in a portion having the highest liquid temperature within the solution becomes lower than or equal to a temperature which is 7° C. lower than a melting point of a solvent in an unfrozen state. ### Claim 31 (depends on 18) The method for treating brain damage according to claim 18, wherein the cell structure is a cell structure obtained by mixing the biocompatible macromolecular blocks and the cells and culturing the mixture for 10 hours or longer. ### Claim 32 (depends on 18) The method for treating brain damage according to claim 18, wherein the cell structure for brain damage treatment is obtained by merging a plurality of the cell structures for brain damage treatment. ### Claim 33 (depends on 18) The method for treating brain damage according to claim 18, wherein the biocompatible macromolecular blocks are blocks of heat-crosslinked polypeptide.