Engineered comestible meat

• US 8,703,216 B2
• Filed: 07/26/2012
• Issued: 04/22/2014
• Est. Priority Date: 07/26/2011
• Status: Active Grant

• Current Assignee: Curators of the University of Missouri (University of Missouri)
• Original Assignee: Curators of the University of Missouri (University of Missouri)
• Inventors: Forgacs, Gabor, Marga, Francoise, Jakab, Karoly Robert
• Primary Examiner(s): Tarazano, D. Lawrence
• Assistant Examiner(s): Anderson, Jerry W
• Application Number:
  US13/558,928
• Publication Number:
  US 20130029008A1
• Time in Patent Office: 635 Days
• Field of Search: 426/55, 426/92, 426/5
• US Class Current: 426/5
• CPC Class Codes: A23L 13/00 Meat products; Meat meal; Preparation or treatment thereof
• CPC Class Codes: A23L 13/03 Coating with a layer; Stuffing, laminating, binding, or compressing of original meat pieces
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12N 2502/1347 Smooth muscle cells
• CPC Class Codes: C12N 2502/28 Vascular endothelial cells
• CPC Class Codes: C12N 2506/1307 from adult fibroblasts
• CPC Class Codes: C12N 2533/76 Agarose, agar-agar
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691

Abstract

Provided are engineered meat products formed as a plurality of at least partially fused layers, wherein each layer comprises at least partially fused multicellular bodies comprising non-human myocytes and wherein the engineered meat is comestible. Also provided are multicellular bodies comprising a plurality of non-human myocytes that are adhered and/or cohered to one another; wherein the multicellular bodies are arranged adjacently on a nutrient-permeable support substrate and maintained in culture to allow the multicellular bodies to at least partially fuse to form a substantially planar layer for use in formation of engineered meat. Further described herein are methods of forming engineered meat utilizing said layers.

Claims

1. A method of forming engineered meat, the method comprising:

preparing a plurality of multicellular bodies comprising a plurality of non-human myocytes cohered to one another;

laying more than one multicellular body adjacently onto a planar support substrate;

fusing said multicellular bodies to form a first planar layer;

stacking about 20 to about 80 additional planar layers onto the first planar layer; and

fusing said layers to form a volume of engineered meat by culturing the stacked planar layers to fuse the planar layers while the planar layers in an inner region of the volume die, so that the majority of cells in the volume have died after fusion between the layers is at least partially complete;

wherein the engineered meat is comestible.

2. The method of claim 1, wherein preparing the plurality of multicellular bodies comprises culturing the plurality of non-human myocyte cells and a plurality of non-human endothelial cells at least until the cells are cohered to one another.

3. The method of claim 1, wherein stacking comprises sequentially stacking the planar layers atop the first planar layer on the planar support substrate.

4. The method of claim 1, wherein culturing the stacked planar layers comprises applying mechanical, electrical or electromechanical force to exercise the myocytes in each layer.

5. The method of claim 1, further comprising freezing said volume of engineered meat.

6. The method of claim 1, wherein preparing the plurality of multicellular bodies comprises preparing a plurality of elongate multicellular bodies comprising a plurality of non-human myocytes cohered to one another and preparing a plurality of substantially spherical multicellular bodies comprising a plurality of non-human myocytes cohered to one another;

and further wherein laying more than one multicellular body comprises laying more than one elongate multicellular body and more than one substantially spherical multicellular body adjacently onto a planar support substrate.

7. The method of claim 6, wherein said elongate multicellular bodies have a length ranging from about 1 mm to about 10 cm.

8. The method of claim 1, wherein preparing the plurality of multicellular bodies comprises preparing a plurality of substantially spherical multicellular bodies comprising a plurality of non-human myocytes cohered to one another;

and further wherein laying more than one multicellular body comprises laying more than one substantially spherical multicellular body adjacently onto a support substrate.

9. The method of claim 1, wherein the planar support substrate is permeable to fluids and nutrients and allows a cell culture media to contact all surfaces of the multicellular bodies.

10. The method of claim 1, wherein said multicellular bodies have a diameter adapted to allow diffusion to sufficiently support the maintenance and growth of said non-human myocytes in culture.

11. The method of claim 1, wherein said multicellular bodies have a diameter of about 100 μ

m to about 500 μ

m.

12. A method of forming a comestible engineered meat product, the method comprising:

forming a plurality of planar layers by adjacently positioning a plurality of multicellular bodies in a plane, wherein each multicellular body comprises a plurality of cohered non-human myocytes;

culturing each of the planar layers at least until the plurality of multicellular bodies within each layer have at least partially fused;

stacking more than about 50 of the plurality of at least partially fused planar layers to form a layered volume of engineered meat; and

culturing the stacked planar layers to fuse the planar layers while the planar layers in an inner region of the volume die, so that the majority of cells in the volume have died after fusion between layers is at least partially complete.