# US 12,180,451 B2 · Packed-bed bioreactor systems and methods of using the same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,180,451 B2 - Document ID: US12180451B2 - Status: Granted - Kind: B2 - Application number: 18/130,668 - Filed: 2023-04-04 - Published: 2023-07-27 - Granted: 2024-12-31 - Earliest claimed: 2019-02-05 - Assignees: Corning Incorporated - Applicants: Corning Incorporated - Inventors: Ann MeeJin Ferrie; Vasiliy Nikolaevich Goral - Web page: https://patentgrep.com/patent/US12180451B2 ## CPC - C12M 25/14: {Scaffolds; Matrices (in general C12N5/0068)} - C12M 23/02: {Form or structure of the vessel (large containers B65D88/00)} - C12M 23/20: {Material Coatings (immunocoatings C12M25/00)} - C12M 23/44: {Multiple separable units; Modules} - C12M 25/04: {in combination with well or multiwell plates, i.e. culture inserts} - C12M 29/10: {Perfusion} - C12M 29/26: {Conditioning fluids entering or exiting the reaction vessel} - C12M 41/26: {of pH} - C12M 25/02: {Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00)} - C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)} - C12N 5/0602: {Vertebrate cells} - C12N 2513/00: 3D culture - C12N 2533/30: Synthetic polymers - C12N 2533/40: Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers - C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue - C12N 2535/00: Supports or coatings for cell culture characterised by topography - 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} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract A bioreactor system is provided that includes a cell culture vessel having a first end, a second end, and at least one reservoir between the first and second ends; and a cell culture matrix disposed in the at least one reservoir. The cell culture matrix has a structurally defined substrate with a surface for adhering cells thereto. The bioreactor system flows material through the at least one reservoir and through the cell culture matrix in a flow direction from the first end to the second end, and the cell culture matrix exhibits isotropic fluid flow permeability therethrough. ## Claims ### Claim 1 A bioreactor system comprising: a cell culture vessel comprising a first end, a second end, and at least one reservoir between the first and second ends; and a cell culture matrix disposed in the at least one reservoir, the cell culture matrix comprising a structurally defined substrate comprising a surface configured for adhering cells thereto, wherein the bioreactor system is configured to flow material through the at least one reservoir and through the cell culture matrix in a flow direction from the first end to the second end, and wherein the cell culture matrix exhibits isotropic fluid flow permeability therethrough. ### Claim 2 (depends on 1) The bioreactor system of claim 1, wherein the cell culture matrix comprises a plurality of sheets of the structurally defined substrate. ### Claim 3 (depends on 2) The bioreactor system of claim 2, wherein the plurality of sheets of the structurally defined substrate are not separated from each other by a spacer material. ### Claim 4 (depends on 1) The bioreactor system of claim 1, wherein the cell culture matrix comprises one or more sheets of the structurally defined substrate, the one or more sheets being disposed in the at least one reservoir in a wound configuration. ### Claim 5 (depends on 4) The bioreactor system of claim 4, wherein the cell culture matrix is disposed within the at least one reservoir such that the wound configuration at least partially surrounds a central longitudinal axis of the cell culture vessel. ### Claim 6 (depends on 1) The bioreactor system of claim 1, wherein the cell culture matrix comprises one or more sheets of the structurally defined substrate, the one or more sheets being disposed in the at least one reservoir as a stack of sheets. ### Claim 7 (depends on 6) The bioreactor system of claim 6, wherein the stack of sheets is arranged perpendicular or parallel to the flow direction. ### Claim 8 (depends on 1) The bioreactor system of claim 1, the cell culture vessel further comprising an inlet at the first end and an outlet at the second end, wherein the bioreactor system is configured to flow the material into the at least one reservoir via the inlet, through the at least one reservoir in the flow direction, and out of the at least one reservoir via the outlet. ### Claim 9 (depends on 1) The bioreactor system of claim 1, wherein the at least one reservoir defines a unitary bulk flow path for material to flow in the flow direction that is substantially parallel to a direction from the first end to the second end. ### Claim 10 (depends on 1) The bioreactor system of claim 1, wherein the structurally defined substrate comprises a plurality of fibers that comprise the surface, the plurality of fibers defining a plurality of pores, wherein the plurality of pores is in an ordered array that is substantially regular and uniform. ### Claim 11 (depends on 10) The bioreactor system of claim 10, wherein the plurality of fibers comprises a first set of fibers arranged parallel to each other in a first fiber direction and a second set of fibers arranged parallel to each other in a second fiber direction, the second fiber direction being different than the first fiber direction. ### Claim 12 (depends on 10) The bioreactor system of claim 10, wherein the plurality of fibers are interwoven. ### Claim 13 (depends on 1) The bioreactor system of claim 1, wherein the substrate comprises a physical structure and a porosity that are in an ordered array that is substantially regular and uniform. ### Claim 14 (depends on 1) The bioreactor system of claim 1, wherein the substrate comprises a rigid monofilament fiber configured to maintain shape under fluid flow. ### Claim 15 (depends on 1) The bioreactor system of claim 1, further comprising means for harvesting the adherent cells or cell by-products from the at least one reservoir. ### Claim 16 (depends on 15) The bioreactor system of claim 15, wherein the cell by-products comprise at least one of proteins, antibodies, viruses, viral vectors, virus-like particles (VLPs), microvessicles, exosomes, and polysaccharides. ### Claim 17 (depends on 1) The bioreactor system of claim 1, wherein the cell culture matrix comprises a plurality of substrates, the plurality of substrates comprising substrates of differing geometries, wherein the differing geometries are different in at least one of a fiber diameter, an opening diameter, or an opening geometry. ### Claim 18 (depends on 1) The bioreactor system of claim 1, wherein the substrate comprises a functionalized surface, the functionalized surface being physically or chemically modified for improved adhesion of the adherent cells to the polymer mesh material. ### Claim 19 (depends on 1) The bioreactor system of claim 1, wherein the cell culture matrix comprises a coating on a surface of the substrate, the coating being configured to promote adherence of the adherent cells. ### Claim 20 (depends on 1) The bioreactor system of claim 1, wherein the at least one reservoir defines a single flow path for material to flow in the flow direction from the first end to the second end.