# US 5,914,264 A · Apparatus for growing vertebrate skin in vitro > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,914,264 A - Document ID: US5914264A - Status: Granted - Kind: A - Application number: 08/903,262 - Filed: 1997-07-25 - Granted: 1999-06-22 - Earliest claimed: 1994-12-30 - Assignees: Reconstructive Technologies, LLC - Inventors: Joshua Korman - Web page: https://patentgrep.com/patent/US5914264A ## CPC - C12N 5/0629: {Keratinocytes; Whole skin} - C12M 21/08: {for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00)} - C12M 35/04: {Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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} ## Abstract A method of growing complete vertebrate skin in vitro, which comprises obtaining a segment of vertebrate skin, positioning the skin segment in an artificial cell-growth medium containing sufficient nutrients to maintain growth of cells of the skin, and subjecting the skin segment to stretching forces while the skin segment is in the medium. Skin produced by the method and an apparatus for carrying out the method are also part of the present invention. ## Claims ### Claim 1 An apparatus for culturing skin tissue, comprising: means for contacting a detached skin segment having a dermal layer side and an epidermal layer side in contact with a nutrient medium on at least the dermal layer side of said segment, and means for stretching said skin while maintaining said contact. ### Claim 2 (depends on 1) The apparatus of claim 1, wherein said apparatus comprises: a container for holding said nutrient medium, at least two connectors for holding said skin segment in said medium at at least two locations on said skin segment, said connectors being attached to said container, and means for supplying opposing forces via said connectors to said skin segment. ### Claim 3 (depends on 2) The apparatus of claim 2, wherein said at least one of said connectors comprises a clamp. ### Claim 4 (depends on 2) The apparatus of claim 2, wherein said means for supplying forces comprises at least one freely suspended weight attached to one of said connectors. ### Claim 5 (depends on 2) The apparatus of claim 2, wherein said means for supplying forces comprises at least one screw adjustment attached to one of said connectors. ### Claim 6 (depends on 2) The apparatus of claim 2, wherein said means for supplying forces comprises at least one electric motor attached to one of said connectors. ### Claim 7 (depends on 1) The apparatus of claim 1, wherein said apparatus comprises: a tubular fluid reservoir having an open end, a clamp located at said open end of said reservoir, wherein said clamp is adapted to seal said skin segment over said open end to provide a fluid-tight seal, and means for supplying hydrostatic pressure to a fluid located in said reservoir. ### Claim 8 (depends on 7) The apparatus of claim 7, wherein means for supplying hydrostatic pressure comprises (1) a nutrient reservoir fluidly connected to said tubular fluid reservoir and being located at a higher gravitational potential than said skin segment when said apparatus is located in its normal operating position or (2) a pump fluidly connected to said tubular reservoir. ### Claim 9 (depends on 2) The apparatus of claim 2, wherein one or more of said connectors comprises a hook. ### Claim 10 (depends on 2) The apparatus of claim 2, wherein one or more of said connectors comprises a suture. ### Claim 11 (depends on 2) The apparatus of claim 2, wherein one or more of said connectors comprises glue. ### Claim 12 (depends on 2) The apparatus of claim 2, wherein said means for supplying forces comprises at least one spring attached to one of said connectors. ### Claim 13 (depends on 4) The apparatus of claim 4, wherein said means for supplying forces comprises at least one weight attached to one of said connectors by a wire, said weight being suspended by a pulley. ### Claim 14 An apparatus for culturing a skin segment having a dermal layer side and an epidermal layer side, comprising: a container for nutrient medium, at least two connectors adapted to hold said skin segment in contact with said nutrient medium on at least the dermal layer side of said segment, said connectors being attached to said container and being capable of moving away from each other while maintaining said contact. ### Claim 15 (depends on 14) The apparatus of claim 14, wherein at least one of said connectors comprises a clamp. ### Claim 16 (depends on 14) The apparatus of claim 14, wherein said at least one of said connectors comprises a hook. ### Claim 17 (depends on 14) The apparatus of claim 14, wherein at least one of said connectors comprises a suture. ### Claim 18 (depends on 14) The apparatus of claim 14, wherein at least one of said connectors comprises glue. ### Claim 19 (depends on 14) The apparatus of claim 14, wherein a force enabling said connectors to move away from one another is applied by at least one freely suspended weight attached to one of said connectors. ### Claim 20 (depends on 14) The apparatus of claim 14, wherein a force enabling said connectors to move away from one another is applied by at least one screw adjustment attached to one of said connectors. ### Claim 21 (depends on 14) The apparatus of claim 14, wherein a force enabling said connectors to move away from one another is applied by at least one spring attached to one of said connectors. ### Claim 22 (depends on 14) The apparatus of claim 14, wherein a force enabling said connectors to move away from one another is applied by at least one electric motor attached to one of said connectors. ### Claim 23 (depends on 14) The apparatus of claim 14, further comprising means for supplying a dynamic stretching force to said skin segment connected to at least one of said connectors. ### Claim 24 (depends on 14) The apparatus of claim 14, further comprising means for supplying a static force to said skin segment connected to at least one of said connectors. ### Claim 25 (depends on 14) The apparatus of claim 14, wherein said apparatus comprises multiple connectors subjected to forces applied in parallel to each other when said connectors are attached to said skin segment. ### Claim 26 (depends on 14) The apparatus of claim 14, wherein said apparatus comprises multiple connectors subjected to forces applied in multiple directions non-parallel to each other in the plane of said skin when said connectors are attached to said skin segment. ### Claim 27 (depends on 14) The apparatus of claim 14, wherein said skin segment has a center and said apparatus comprises multiple connectors subjected to forces applied radially outward from said center when said connectors are attached to said skin segment.