# US 10,500,306 B2 · Method and device for producing a bioartificial tissue construct > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,500,306 B2 - Document ID: US10500306B2 - Status: Granted - Kind: B2 - Application number: 14/367,247 - Filed: 2012-12-20 - Published: 2014-11-20 - Granted: 2019-12-10 - Earliest claimed: 2011-12-23 - Assignees: Medizinische Hochschule Hannover - Applicants: Medizinische Hockschule Hannover - Inventors: Thomas Aper; Mathias Wilhelmi; Axel Haverich - Web page: https://patentgrep.com/patent/US10500306B2 ## CPC - A61L 27/225: {Fibrin; Fibrinogen} - A61F 2/062: {Apparatus for the production of blood vessels made from natural tissue or with layers of living cells} - A61F 2/07: Stent-grafts - A61F 2/90: characterised by a net-like or mesh-like structure - A61L 27/06: Titanium or titanium alloys - A61L 27/3804: {characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells} - A61L 27/3808: {Endothelial cells} - A61L 27/3826: {Muscle cells, e.g. smooth muscle 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 27/507: {for artificial blood vessels (apparatus for applying cells on a blood vessel prosthesis A61F2/062)} - B29C 41/04: Rotational or centrifugal casting, i.e. coating the inside of a mould by rotating the mould - B29C 41/36: Feeding the material on to the mould, core or other substrate - B29C 41/38: Moulds, cores or other substrates - C12M 21/08: {for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00)} - C12M 23/48: {Holding appliances; Racks; Supports (holding devices for laboratory apparatus B01L9/00)} - B29K 2101/00: Use of unspecified macromolecular compounds as moulding material {(use of unspecified rubbers B29K2021/00)} - B29K 2901/00: Use of unspecified macromolecular compounds as mould material (use of unspecified rubbers B29K2821/00) - B29K 2905/00: Use of metals, their alloys or their compounds, as mould material - B29L 2031/753: {Medical equipment; Accessories therefor (bloodbags, medical bags B29L2031/7148; artificial eyes B29L2011/0008)} - 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) - A61F: FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS (dental prosthetics A61C) - B29C: SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING (making preforms B29B11/00; making laminated products by combining previously unconnected layers which become one product whose layers will remain together B32B37/00 - B32B41/00) - 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} - B29K: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES B29B, B29C OR B29D, RELATING TO MOULDING MATERIALS OR TO MATERIALS FOR {MOULDS, } REINFORCEMENTS, FILLERS OR PREFORMED PARTS, e.g. INSERTS - B29L: INDEXING SCHEME ASSOCIATED WITH SUBCLASS B29C, RELATING TO PARTICULAR ARTICLES ## Abstract The tissue construct with viable cells in an extracellular matrix made of fibrin is produced with a special method, in which a matrix material and cells are shaped into a hollow body, in particular a tubular hollow body, by means of a rotational casting method in a hollow mould (1), the method comprising the following steps: (a) introduction of cells of at least one cell type and/or a fibrinogen preparation into the rotating hollow mould (1) with the aid of an applicator (4), said applicator (4) being displaced along the rotational axis during the introduction and step (a) being performed one or more times; (b) continuation of the rotation process until the fibrinogen solidifies into a dimensionally stable matrix, obtaining a primarily solidified tissue construct; (c) removal of the tissue construct from the mould. The construct can also he obtained in a relatively short time from autologous materials. ## Claims ### Claim 1 A method for producing a tissue construct containing viable cells in an extracellular matrix, in which a matrix material and cells are molded by means of a rotary casting method in a hollow mold to give a hollow body, having the following steps: (a) introducing cells of at least one cell type together with a fibrinogen preparation in a mixture with the aid of an applicator into the rotating hollow mold, wherein the applicator during the introduction is shifted along the axis of rotation and wherein step (a) is carried out once or several times; (b) continuing rotation of the hollow mold up to solidification of the fibrinogen to form a dimensionally stable matrix, obtaining a primary-solidified tissue construct; and (c) demolding the tissue construct, wherein the cells are introduced while a rotation of 100 g to 650 g is carried out. ### Claim 2 (depends on 1) The method as claimed in claim 1, wherein a plurality of cell types are used simultaneously or sequentially. ### Claim 3 (depends on 2) The method as claimed in claim 2, whereby successively following application steps according to step (a), fibroblasts, smooth muscle cells and endothelial cells are used, or in one or more application steps according to step (a), only adipose tissue derived stern cells (ASC), or ASC in a mixture with endothelial cells, are used. ### Claim 4 (depends on 1) The method as claimed in claim 1, wherein the cells are applied in each case in a mixture with the fibrinogen preparation, or simultaneously with the fibrinogen preparation, or in a mixture with a component of the fibrinogen preparation. ### Claim 5 (depends on 1) The method as claimed in claim 1, wherein the cells and the fibrinogen preparation are introduced by spraying. ### Claim 6 (depends on 1) The method as claimed in claim 1, wherein, in the hollow mold, before the start of the introduction of cells and fibrinogen, or between a plurality of steps according to step (a), in addition a support frame is used. ### Claim 7 (depends on 1) The method as claimed in claim 1, wherein a post-treatment of the primary-solidified tissue construct is performed in a bioreactor. ### Claim 8 (depends on 1) The method as claimed in claim 1, wherein the hollow body is a tubular hollow body. ### Claim 9 (depends on 2) The method according to claim 2, wherein the cell types are selected from the group of fibroblasts, fibrocytes, muscle cells, endothelial cells (EC) and cells obtained from fat tissue. ### Claim 10 (depends on 9) The method according to claim 9, wherein the cell types used include muscle cells selected from SMC and SPC. ### Claim 11 (depends on 9) The method as claimed in claim 9, wherein the cell types used include endothelial cells selected from EPC and EOEC. ### Claim 12 (depends on 9) The method as claimed in claim 9, wherein the cell types used include cells obtained from fat tissue, wherein said cells obtained from fat tissue are ASC. ### Claim 13 (depends on 4) The method as claimed in claim 4, wherein the mixture with the cells are applied in each case in a mixture with a fibrinogen cross linking agent. ### Claim 14 (depends on 5) The method as claimed in claim 5, wherein a layer thickness of the cells and the fibrinogen preparation is ≤1 mm. ### Claim 15 (depends on 5) The method as claimed in claim 5, wherein a layer thickness of the cells and the fibrinogen preparation is ≤0.5 mm. ### Claim 16 (depends on 6) The method as claimed in claim 6, wherein the support frame is made of metal or plastic. ### Claim 17 (depends on 6) The method as claimed in claim 6, wherein the support frame is in the form of a grating. ### Claim 18 (depends on 13) The method as claimed in claim 13, wherein an inner surface of the hollow mold is coated with polytetrafluoroethylene or includes a detachable polytetrafluoroethylene insert. ### Claim 19 (depends on 1) The method as claimed in claim 1, wherein the cells are present on a side of a layer of the matrix facing the hollow mold. ### Claim 20 (depends on 1) The method as claimed in claim 1, further comprising the step of lining the hollow mold with a support grid or support frame prior to performing steps a)-c).