# US 11,993,767 B2 · Method for producing 3D, biocompatible polymer scaffold with a cell-filled cavity > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,993,767 B2 - Document ID: US11993767B2 - Status: Granted - Kind: B2 - Application number: 17/779,100 - Filed: 2020-11-20 - Published: 2023-05-04 - Granted: 2024-05-28 - Earliest claimed: 2019-11-27 - Assignees: CELLBRICKS GmbH - Applicants: CELLBRICKS GmbH - Inventors: Lutz Kloke; Alexander Thomas; Tobias Lam - Web page: https://patentgrep.com/patent/US11993767B2 ## CPC - C12M 25/14: {Scaffolds; Matrices (in general C12N5/0068)} - B29C 64/124: using layers of liquid which are selectively solidified - B29C 64/135: the energy source being concentrated, e.g. scanning lasers or focused light sources - B33Y 10/00: Processes of additive manufacturing - B33Y 70/00: Materials specially adapted for additive manufacturing - B33Y 80/00: Products made by additive manufacturing - B81C 1/00119: {Arrangement of basic structures like cavities or channels, e.g. suitable for microfluidic systems} - C12M 33/00: {Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus (chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00)} - C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)} - B81B 2201/051: Micromixers, microreactors - B81B 2201/058: Microfluidics not provided for in B81B2201/051 - B81B2201/054 - B81B 2203/0315: Cavities - C12N 2513/00: 3D culture - C12N 2537/10: Cross-linking - 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} - 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) - B33Y: ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING - B81C: PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS (making microcapsules or microballoons B01J13/02; processes or apparatus specially adapted for the manufacture or treatment of piezoelectric, electrostrictive or magnetostrictive elements per se H10N30/01) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - B81B: MICROSTRUCTURAL DEVICES OR SYSTEMS, e.g. MICROMECHANICAL DEVICES (piezoelectric, electrostrictive or magnetostrictive elements per se H10N30/00, H10N35/00) ## Abstract A 3D scaffold of a biocompatible polymer and colonized with biological cells is provided. The biological cells can be cultured to form a 3D cell culture construct that closely approximates a physiological architecture. A method for producing the 3D scaffold colonized with biological cells is also provided. ## Claims ### Claim 1 A method including: (a) constructing a 3D scaffold comprising a biocompatible polymer using a lithographic 3D printing method, the 3D scaffold having an at least partially covered cavity and wherein the biocompatible polymer is obtained by rendering a biopolymer as a starting substance photopolymerizable or photocrosslinkable by introducing a photoreactive group, the biopolymer being selected from a group consisting of proteins, DNA, RNA, carbohydrates, carbohydrate derivatives, collagens, fibrins, alginates, gelatins, and hyaluronic acids; and (b) filling the at least partially covered cavity with a suspension containing biological cells to colonize the 3D scaffold. ### Claim 2 (depends on 1) The method of claim 1 wherein the 3D scaffold is constructed using a stereolithographic 3D printing method. ### Claim 3 (depends on 1) The method of claim 1 wherein constructing the 3D scaffold includes curing a first photopolymerizable or photocrosslinkable substance by focusing electromagnetic radiation in a first focal plane in which the first photopolymerizable or photocrosslinkable substance is present. ### Claim 4 (depends on 3) The method of claim 3 wherein constructing the 3D scaffold includes curing a second photopolymerizable or photocrosslinkable substance by focusing electromagnetic radiation in a second focal plane in which the second photopolymerizable or photocrosslinkable substance is present, the second focal plane being different from the first focal plane. ### Claim 5 (depends on 4) The method of claim 4 wherein the second photopolymerizable or photocrosslinkable substance is different from the first photopolymerizable or photocrosslinkable substance. ### Claim 6 (depends on 1) The method of claim 1 wherein filling the at least partially covered cavity with the suspension containing biological cells includes directing the suspension containing biological cells through a filling opening on the 3D scaffold. ### Claim 7 (depends on 1) The method of claim 1 further including drying the 3D scaffold prior to filling the at least partially covered cavity with the suspension containing biological cells. ### Claim 8 (depends on 1) The method of claim 1 further including, after filling the at least partially covered cavity with the suspension containing biological cells, culturing the biological cells to form a 3D cell culture construct. ### Claim 9 (depends on 8) The method of claim 8 wherein the 3D cell culture construct is penetrated by further cells, viruses, bacteria, enzymes or active substances. ### Claim 10 A 3D scaffold comprising: (a) multiple layers of a biocompatible polymer, the multiple layers of the biocompatible polymer formed by a lithographic 3D printing method and defining an at least partially covered cavity and wherein the biocompatible polymer is obtained by rendering a biopolymer as a starting substance photopolymerizable or photocrosslinkable by introducing a photoreactive group, the biopolymer being selected from a group consisting of proteins, DNA, RNA, carbohydrates, carbohydrate derivatives, collagens, fibrins, alginates, gelatins, and hyaluronic acids; and (b) a suspension containing biological cells filling the at least partially covered cavity to colonize the 3D scaffold. ### Claim 11 (depends on 10) The 3D scaffold of claim 10 further including one or more outlet openings. ### Claim 12 (depends on 10) The 3D scaffold of claim 10 further including columns, grids, or crosspieces in the at least partially covered cavity. ### Claim 13 (depends on 10) The 3D scaffold of claim 10 wherein at least a portion of a first one of the multiple layers of the biocompatible polymer comprises a first photopolymerizable or photocrosslinkable substance cured by focusing electromagnetic radiation in a first focal plane in which the first photopolymerizable or photocrosslinkable substance is present. ### Claim 14 (depends on 13) The 3D scaffold of claim 13 wherein at least a portion of the first one of the multiple layers of the biocompatible polymer or a second one of the multiple layers of the biocompatible polymer comprises a second photopolymerizable or photocrosslinkable substance cured by focusing electromagnetic radiation in a second focal plane in which the second photopolymerizable or photocrosslinkable substance is present, the second focal plane being different from the first focal plane. ### Claim 15 (depends on 14) The 3D scaffold of claim 14 wherein the second photopolymerizable or photocrosslinkable substance is different from the first photopolymerizable or photocrosslinkable substance. ### Claim 16 (depends on 10) The 3D scaffold of claim 10 wherein the biological cells are cultured to form a 3D cell culture construct. ### Claim 17 (depends on 16) The 3D scaffold of claim 16 wherein the 3D cell culture construct is penetrated by further cells, viruses, bacteria, enzymes or active substances.