# US 2025/0367348 A1 · 3d printing of 3-layered retina and choroid tissue > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2025/0367348 A1 - Document ID: US20250367348A1 - Status: Published application - Kind: A1 - Application number: 19/303,562 - Filed: 2025-08-19 - Published: 2025-12-04 - Earliest claimed: 2023-02-19 - Assignees: Ramot at Tel-Aviv University Ltd.; Ichilov Tech Ltd. - Applicants: Ramot at Tel-Aviv University Ltd.; Ichilov Tech Ltd. - Inventors: Tal DVIR; Yahel SHECHTER; Eric SILBERMAN; Aya BARZELAY WOLLMAN; Adiel BARAK - Web page: https://patentgrep.com/patent/US20250367348A1 ## CPC - A61L 27/3808: {Endothelial cells} - A61L 27/3633: {Extracellular matrix [ECM]} - A61L 27/3813: {Epithelial cells, e.g. keratinocytes, urothelial cells} - A61L 27/52: Hydrogels or hydrocolloids - B33Y 40/20: Post-treatment, e.g. curing, coating or polishing - B33Y 70/00: Materials specially adapted for additive manufacturing - B33Y 80/00: Products made by additive manufacturing - C12N 5/062: {Sensory transducers, e.g. photoreceptors; Sensory neurons, e.g. for hearing, taste, smell, pH, touch, temperature, pain} - C12N 5/0621: {Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062)} - C12N 5/0697: {Artificial constructs associating cells of different lineages, e.g. tissue equivalents (blood vessels C12N5/0691)} - A61L 2430/16: for reconstruction of eye parts, e.g. intraocular lens, cornea - B29C 64/112: using individual droplets, e.g. from jetting heads - B33Y 10/00: Processes of additive manufacturing - C12N 2513/00: 3D culture - C12N 2533/30: Synthetic polymers - C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue - C12N 2537/10: Cross-linking - 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) - 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 - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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) ## Abstract A method for reinforcing a cellularized retinal construct fabricated from (i) endothelial cells; (ii) retinal pigment epithelial cells and/or photoreceptors; and (iii) an extracellular matrix (ECM) hydrogel is disclosed. The method comprises contacting the construct with a biocompatible small-molecule reinforcing agent that is capable of chemically interacting with the ECM hydrogel under conditions that maintain viability of the cells, to thereby increase a compressive modulus of the ECM hydrogel by at least 10%. ## Claims ### Claim 1 A method for reinforcing a cellularized retinal construct fabricated from: (i) endothelial cells; (ii) retinal pigment epithelial (RPE) cells and/or photoreceptors; and (iii) an extracellular matrix (ECM) hydrogel, the method comprising: contacting the cellularized retinal construct with a biocompatible small-molecule reinforcing agent that is capable of chemically interacting with the ECM hydrogel under conditions that maintain viability of the cells, to thereby increase a compressive modulus of the ECM hydrogel by at least 10%. ### Claim 2 (depends on 1) The method of claim 1, wherein said chemically interacting effects cross-linking of the ECM hydrogel. ### Claim 3 (depends on 1) The method of claim 1, wherein said reinforcing agent is a polyaldehyde. ### Claim 4 (depends on 1) The method of claim 1, wherein said reinforcing agent is an oxidized, poly-aldehyde saccharide. ### Claim 5 (depends on 1) The method of claim 1, wherein said contacting is with a culturing medium that comprises said reinforcing agent. ### Claim 6 (depends on 5) The method of claim 5, wherein said reinforcing agent is an oxidized, poly-aldehyde saccharide and wherein a concentration of said reinforcing agent in said medium is less than 0.1% by weight. ### Claim 7 (depends on 1) The method of claim 1, further comprising generating the cellularized retinal construct prior to the contacting by sequentially forming a plurality of layers on a receiving medium, wherein a first of said layers comprises said endothelial cells and a second of said layers comprises said RPE cells or said photoreceptors. ### Claim 8 (depends on 7) The method according to claim 7, wherein said second layer comprises RPE cells and a third layer comprises said photoreceptors. ### Claim 9 (depends on 7) The method of claim 7, further comprising culturing said cellularized retinal construct for at least 3 days following said generating and prior to said contacting. ### Claim 10 (depends on 1) The method of claim 1, wherein said ECM hydrogel is generated from decellularized omentum. ### Claim 11 A method of generating an engineered cellularized retinal construct comprising: (a) culturing endothelial cells in the presence of an ECM hydrogel derived from decellularized omental tissue to generate a layer of endothelial cells; (b) culturing RPE cells on said layer of endothelial cells to generate a layer of RPE cells; and (c) culturing photoreceptor cells on said layer of RPE cells, thereby generating the cellularized retinal construct. ### Claim 12 (depends on 11) The method according to claim 11, further comprising contacting the retinal construct with a biocompatible small-molecule reinforcing agent that is capable of chemically interacting with the ECM hydrogel under conditions that maintain viability of the cells, to thereby increase a compressive modulus of the ECM hydrogel by at least 10%. ### Claim 13 (depends on 12) The method of claim 12, wherein said reinforcing agent is a polyaldehyde. ### Claim 14 (depends on 12) The method of claim 12, wherein said reinforcing agent is an oxidized, poly-aldehyde saccharide. ### Claim 15 (depends on 12) The method of claim 12, wherein said contacting is with a culturing medium that comprises said reinforcing agent. ### Claim 16 (depends on 1) A cellularized, engineered retinal construct generated according to the method of claim 1. ### Claim 17 A cellularized engineered retinal construct comprising endothelial cells and retinal cells distributed within a chemically cross-linked ECM hydrogel, wherein said ECM hydrogel is chemically cross-linked by a biocompatible small-molecule reinforcing agent that is capable of chemically interacting with the ECM hydrogel under conditions that maintain viability of the cells, and wherein a compressive modulus of the ECM hydrogel is higher by at least 50% than a compressive modulus of the ECM hydrogel which is not chemically cross-linked. ### Claim 18 (depends on 1) A method of treating a disease or condition associated with a damaged retina in a subject in need thereof, the method comprising implanting the cellularized construct of claim 1 into the subject, thereby treating the condition.