权利要求:
What is claimed:
1. A computerized bio-printing method for forming an edible biostructure having
predetermined 3D pattern of cells therein, the method comprising: a. providing a bio-printing system comprising: i. a microprocessor in communication with: a non-volatile storage device having thereon a microprocessor-readable medium with set of executable instructions configured to, when executed, to cause the microprocessor to execute the method of bio-printing bio printing; and an image library corresponding to a two dimensional (2D) layer of at least one of: cells, scaffolding, and connective tissue, within the biostructure having predetermined 3D pattern of cells therein; ii. a first bio-ink dispenser in communication with the microprocessor having: at least one aperture, a first bio-ink reservoir, and a first actuator operable to dispense the first bio ink through the aperture, wherein the first bio-ink is a composition comprising viable cells suspended in at least one of: a bio-compatible dispersing medium and bio-compatible polymer, monomer, or oligomer; iii. a second dispenser in communication with the microprocessor having: at least one aperture, a second bio-ink reservoir, and a second actuator operable to dispense the second bio-ink through the aperture, wherein the second bio-ink is a composition comprising bio compatible compounds suspended at least one of: a bio-compatible dispersing medium and bio-compatible polymer, monomer, or oligomer; iv. a third dispenser in communication with the microprocessor having: at least one aperture, a third bio-ink reservoir, and a third actuator operable to dispense the third bio-ink through the aperture, wherein the third bio-ink comprises means for functionalizing the bio compatible polymer, monomer, or oligomer; and v. a conveyor in communication with the microprocessor, operably coupled to each of the first, the second, and the third actuators;
b. uploading from the library a first image corresponding to a first two dimensional (2D) layer of the edible biostructure having predetermined 3D pattern of cells; c. using the first dispenser, forming a first layer of bio-ink pattern or portion thereof; d. using the third dispenser, functionalizing the first layer of bio-ink pattern or its portion; e. using the second dispenser, forming a first layer of predetermined pattern of the second bio-ink composition; and f. using the third dispenser, functionalizing the first layer of predetermined pattern of the second bio-ink.
2. The method of claim 1, wherein the step of functionalizing the first bio-ink layer pattern and/or the step of functionalizing the second bio-ink layer pattern comprises depositing ionic solution, cross linking, exposing the cells to electromagnetic radiation, or a combination comprising one or more of the foregoing.
3. The method of any one of claim 2, wherein the first bio-ink composition, the second bio-ink composition or both comprises: collagen, fibrin, laminin, PEGilated-methacrylate, chitosan, carrageenan, poly(lactic) acid, poly(lactic-co-glycolic acid), their methacrylate conjugates, co polymers, interpenetrating networks or a composition comprising one or more of the foregoing.
4. The method of any one of claims 3, wherein the first bio-ink comprises one or more of endothelial cells, muscle cells, fibroblast cells, mesothelial cells, pericyte cells, monocyte cells, plasma cells, mast cells, adipocyte cells, chondrocyte cells, osteoblasts, osteoclasts, osteocytes, cells population manipulated from a specific type of cell to another, or cells composition comprising one or more of the foregoing cells; a bio-compatible polymer; and optionally a photoinitiator.
5. The method of claim 3, wherein the carrageenan is kappa carrageenan and functionalizing comprises heating and using the third bio-ink dispenser; depositing ionic solution comprising KC1, CaCk, or a composition comprising one or both of the foregoing.
6. The method of claim 3, wherein the first bio-ink composition comprises PEGilated- methacrylate and functionalizing comprises exposure to electromagnetic radiation.
7. The method of any one of claims 1, wherein following functionalization, the biostructure exhibits compressive modulus is no less than 0.08 MPa.
8. The method of any one of claims 1, wherein an extracellular matrix composition included in the second bio-ink comprises a composition configured to accelerate cell adhesion, promote cell colonization, proliferation, or manipulation, retain native growth of the cells, and/or organ and/or tissue.
9. The method of Claim 9, wherein the composition comprises epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), bone morphogenic protein (BMP), insulin-like growth factor (IGF), glucoseaminoglycan (GAG), Transforming growth factor (TGF) or extracellular matrix compound composition comprising the foregoing.
10. The method of claim 1, further comprising, subsequent to the step of functionalizing the first layer of predetermined pattern of the second bio-ink: a. uploading from the library a subsequent image corresponding to a second or subsequent two dimensional (2D) layer or portion thereof of bio-ink within the biostmcture having predetermined 3D pattern of cells; b. using the first dispenser, forming the second layer of bio-ink pattern or portion thereof on and/or adjacent to the first layer of the first bio-ink pattern, and/or the first layer of predetermined pattern of the second bio-ink composition; and c. functionalizing the second layer of bio-ink pattern.
11. The method of claim 10, further comprising: a. using the second dispenser, forming a second layer or portion thereof of
predetermined pattern of the second bio-ink on, and/or adjacent to the first layer of first bio-ink pattern and/or the first layer of predetermined pattern of the second bio-ink composition; and b. functionalizing the second layer of predetermined pattern of the second bio-ink.
12. The method of claim 11, wherein the cells in the second layer of bio-ink pattern is different than the first layer of cells.
13. The method of claim 12, wherein the bio-compatible compounds in the second layer of predetermined pattern of the second bio-ink, is different than the bio-compatible compounds in the first layer of predetermined pattern of the second bio-ink.
14. The method of any one of claims 1, wherein the biostmcture is at least one of: a tissue, an organ, a tissue scaffold, and a connective tissue.
15. The method of claim 14, wherein the predetermined 3D pattern of the first bio-ink in the biostructure is non-random.
16. The method of claim 14, further comprising a step of transferring the edible biostmcture to a bio-reactor; and growing the biostmcture.
17. The method of claim 14, wherein the tissue scaffold is sized and adapted to maintain liquid communication among cells within the biostmcture.
18. An edible food product fabricated using the method of claim 1.
19. The edible food product of claim 18, wherein the biostmcture is configured to emulate a bovine muscle tissue, a porcine muscle tissue, an avian muscle tissue, agnatha muscle tissue, chrondrichthyes muscle tissue, osteichthyes muscle tissue, and caprine muscle tissue.