# US 12,180,449 B2 · Self-contained mold for culture and transport of 3D cellular gels > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,180,449 B2 - Document ID: US12180449B2 - Status: Granted - Kind: B2 - Application number: 16/284,449 - Filed: 2019-02-25 - Published: 2019-10-10 - Granted: 2024-12-31 - Earliest claimed: 2018-04-06 - Assignees: Boise State University - Applicants: Boise State University - Inventors: John Everingham; Trevor Lujan - Web page: https://patentgrep.com/patent/US12180449B2 ## CPC - C12M 23/34: {Internal compartments or partitions} - C12M 25/14: {Scaffolds; Matrices (in general C12N5/0068)} - B01L 3/502707: {characterised by the manufacture of the container or its components} - B01L 3/505: {Flexible containers without fluid transport within} - C12M 21/08: {for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00)} - C12M 23/02: {Form or structure of the vessel (large containers B65D88/00)} - C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)} - C12N 2527/00: Culture process characterised by the use of mechanical forces, e.g. strain, vibration - C12N 2533/40: Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers - C12N 2535/00: Supports or coatings for cell culture characterised by topography - C12N 2539/00: Supports and/or coatings for cell culture characterised by properties - 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} - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract A mold capable of preparing a cellular or acellular construct includes a block including a three-dimensional cavity for a sample, a base, and a removable sheet inserted between the block and the base. The sheet may be coated with silicone grease prior to insertion of the sheet between the block and the base. Mechanical constraints may be included at each end of the cavity to constrain cell driven contraction and to allow for gripping the sample after the sample is removed from the mold. The sample may be prepared by partially filling the cavity with gel, filling the remaining portion of the cavity with culture media, and allowing a three-dimensional sample to culture within the cavity for a time. The sample may be transported to a bioreactor and ejected from the mold by removing the sheet and applying pressure to the mechanical constraints. ## Claims ### Claim 1 A system for preparing and transporting a cellular or acellular construct for controlled extraction of the construct, the system comprising: a sample constructed within a three-dimensional cellular scaffold and culture media; a block including a block height of at least twenty millimeters, a block width that is more than the block height, a block length that is more than the block width, a three-dimensional cavity for constructing the sample in three dimensions, wherein the block is made from a bio-inert material that does not produce an adverse effect with said sample, wherein the three-dimensional cavity comprises: a cavity height that is identical to the block height, a first sub-cavity having a first mechanical constraint, a second sub-cavity having a second mechanical constraint, a third, elongated sub-cavity, wherein the first sub-cavity and the second sub-cavity are located at each end of the third, elongated sub-cavity, and a dog-bone and/or cruciform shape; a base including a base height that is less than the block height, wherein the base is made from a bio-inert material; and an impermeable, removable sheet inserted into a cutout in the base and slid beneath the block. ### Claim 2 (depends on 1) The system of claim 1, wherein the block is made from polytetrafluoroethylene (PTFE). ### Claim 3 (depends on 1) The system of claim 1, wherein the base is made from poly lactic acid (PLA). ### Claim 4 (depends on 1) The system of claim 1, wherein the impermeable, removable sheet is made from polytetrafluoroethylene (PTFE). ### Claim 5 (depends on 1) The system of claim 1, wherein the impermeable, removable sheet is thinner than the base. ### Claim 6 (depends on 1) The system of claim 1, wherein the impermeable, removable sheet is coated with silicone grease prior to insertion of the impermeable, removable sheet between the block and the base. ### Claim 7 (depends on 1) The system of claim 1, the block width is thirty-three millimeters, and the block length is fifty-seven millimeters. ### Claim 8 (depends on 7) The system of claim 7, wherein the base has a base width identical to the block width, a base length identical to the block length, and the base height is one and a half millimeters. ### Claim 9 (depends on 1) The system of claim 1, wherein the base includes a rectangular cut-out spanning an entire thickness of the base and a rectangular ridge supporting the impermeable, removable sheet. ### Claim 10 (depends on 9) The system of claim 9, wherein the rectangular ridge has a rectangular ridge height of one millimeter and a rectangular ridge width of at least one millimeter. ### Claim 11 (depends on 1) The system of claim 1, wherein the cavity has a sample width of seven millimeters and a gauge length of twenty-one millimeters. ### Claim 12 (depends on 1) The system of claim 1, wherein the first and second mechanical constraints are positioned at each end of the three-dimensional cavity to constrain cell driven contraction and to allow for gripping the sample after the sample is removed from the mold. ### Claim 13 (depends on 1) The system of claim 1, wherein the first constraint and/or the second constraint comprises a natural fiber tab. ### Claim 14 (depends on 1) The system of claim 1, wherein the first constraint and/or the second constraint comprise a porous polyethylene bar. ### Claim 15 (depends on 1) The system of claim 1, wherein the three-dimensional cellular scaffold comprises a collagen gel. ### Claim 16 (depends on 15) The system of claim 15, wherein the sample comprises less than forty percent of the collagen gel and more than sixty percent of the culture media. ### Claim 17 (depends on 1) The system of claim 1, wherein the three-dimensional cellular scaffold comprises fibrin. ### Claim 18 (depends on 1) The system of claim 1, wherein the three-dimensional cellular scaffold comprises agarose. ### Claim 19 (depends on 1) The system of claim 1, wherein the three-dimensional cellular scaffold comprises a hydrogel.