Apparatuses and methods for preparing a comestible meat product

Patent metadata

Field	Value
Publication number	US-11357244-B2
Application number	US-202017131514-A
Country	US
Kind code	B2
Filing date	Dec 22, 2020
Priority date	May 28, 2019
Publication date	Jun 14, 2022
Grant date	Jun 14, 2022

Abstract

The substrates, systems, and methods described herein relate to textured substrates for preparing a comestible meat product. Substrates and methods are described herein for controlling one or more of growth, adhesion, retention, and/or release of cells (e.g., of a cell sheet) on or from the surface of the substrate. A method of preparing a comestible meat product may include applying a plurality of non-human cells to at least one patterned texture substrate, growing the cells on the patterned texture substrate to form the comestible meat product, and separating the comestible meat product from the patterned texture substrate. The patterned texture allows for improved growth, adhesion, retention, and/or release of cells as compared to another surface not comprising the patterned texture. In some embodiments, the cell culture substrate surfaces include a plurality of regions corresponding to a plurality of patterned textures.

First claim

We claim:

1. A method of preparing a comestible meat product, comprising:
applying a plurality of non-human cells comprising at least one muscle-derived cell and one or more additional cells, at any state of differentiation, from at least one of myoblasts, mesoangioblasts, myofibroblasts, myotubes, mesenchymal stem cells, hepatocytes, fibroblasts, pericytes, adipocytes, adipose tissue, epithelial, chondrocytes, osteoblasts, osteoclasts, pluripotent cells, somatic stem cells, endothelial cells, epithelial tissue, vascular endothelium, or contractile cells, to a patterned texture substrate comprising a first patterned texture region and a second patterned texture region, wherein a texture of the first patterned texture region differs from a texture of the second patterned texture region;
directing a cell culture medium continuously at a first fluid flow rate across or adjacent to the patterned texture substrate;
growing the plurality of non-human cells on the patterned texture substrate to form the comestible meat product; and
separating the comestible meat product from the patterned texture substrate.

2. The method of claim 1 , wherein the comestible meat product is held intact by an extracellular matrix secreted by the plurality of non-human cells.

3. The method of claim 1 , wherein the comestible meat product is separated contemporaneously as a cell sheet from the patterned texture substrate.

4. The method of claim 1 , further comprising positioning the patterned texture substrate in a bioreactor.

5. The method of claim 1 , wherein the plurality of non-human cells is retained on the patterned texture substrate for at least 3 days.

6. The method of claim 1 , wherein the plurality of non-human cells comprise one or more cells from at least one of livestock, poultry, game, or aquatic animal species.

7. The method of claim 1 , wherein the plurality of non-human cells comprise one or more cells from at least one of a vertebrate or invertebrate species.

8. The method of claim 1 , wherein the first patterned texture region or the second patterned texture region comprises one or more of channels, recesses, or protrusions.

9. The method of claim 1 , wherein the texture of the first patterned texture region or the texture of the second patterned texture region comprises a plurality of surface characteristics that correspond to a cell type or at least one of growth, retention, or release of the plurality of non-human cells.

10. The method of claim 1 , wherein the first patterned texture region or the second patterned texture region comprises a set of linear channels in parallel to each other.

11. The method of claim 1 , wherein the texture of the first patterned texture region or the texture of the second patterned texture region comprises a channel having a predetermined direction comprising one or more of a radial, circular, non-linear, spot, or cross-hatched pattern.

12. The method of claim 1 , wherein the texture of the first patterned texture region or the texture of the second patterned texture region comprises a random spatial distribution pattern.

13. The method of claim 1 , wherein the patterned texture substrate is arranged in a three-dimensional lattice with one or more additional substrates.

14. The method of claim 1 , wherein the patterned texture substrate is placed into or integral with a culture vessel, wherein the culture vessel comprises one or more of a bioreactor, well, petri dish, plate, flask, cylinder, tank, box, or fixed surface.

15. The method of claim 1 wherein the patterned texture substrate comprises a biocompatible coating.

16. The method of claim 1 , wherein directing the cell culture medium comprises directing the cell culture medium in a fluid flow direction based on at least one of the texture of the first patterned texture region or the texture of the second patterned texture region.

17. The method of claim 1 , wherein the patterned texture substrate comprises a non-patterned texture region.

18. The method of claim 17 , wherein the cell culture medium is directed to the non-patterned texture region.

19. A method of preparing a comestible meat product, the method comprising:
applying a plurality of non-human cells comprising at least one muscle-derived cell and one or more additional cells, at any state of differentiation, from at least one of myoblasts, mesoangioblasts, myofibroblasts, myotubes, mesenchymal stem cells, hepatocytes, fibroblasts, pericytes, adipocytes, adipose tissue, epithelial, chondrocytes, osteoblasts, osteoclasts, pluripotent cells, somatic stem cells, endothelial cells, epithelial tissue, vascular endothelium, or contractile cells to at least one patterned texture substrate;
growing the plurality of non-human cells on the at least one patterned texture substrate to form a comestible meat product, wherein an arrangement of patterned textures of the at least one patterned texture substrate inhibits release of the plurality of non-human cells for a predetermined amount of time or until the plurality of non-human cells achieve a predetermined cell state; and
separating the comestible meat product from the at least one patterned texture substrate as a substantially intact cell sheet using fluidic shear stress.

20. A method of preparing a comestible meat product, the method comprising:
applying a plurality of non-human cells comprising at least one muscle-derived cell and one or more additional cells, at any state of differentiation, from at least one of myoblasts, mesoangioblasts, myofibroblasts, myotubes, mesenchymal stem cells, hepatocytes, fibroblasts, pericytes, adipocytes, adipose tissue, epithelial, chondrocytes, osteoblasts, osteoclasts, pluripotent cells, somatic stem cells, endothelial cells, epithelial tissue, vascular endothelium, or contractile cells to at least one cell-adherent patterned texture of a patterned texture substrate;
growing the plurality of non-human cells applied to the patterned texture substrate to form a comestible meat product such that the plurality of non-human cells have a cell alignment based on the at least one cell-adherent patterned texture substrate; and
separating the comestible meat product from the patterned texture substrate using a fluid flow in a predetermined direction according to an adhesion gradient of adhesion quality between the comestible meat product and the patterned texture substrate.

21. The method of claim 1 , wherein directing the cell culture medium comprises directing the cell culture medium according to a predetermined flow pattern.

22. The method of claim 21 , wherein the predetermined flow pattern comprises a predetermined flow path over the patterned texture substrate.

23. The method of claim 1 , wherein separating the comestible meat product comprises directing a fluid at the patterned texture substrate at a second fluid flow rate higher than the first fluid flow rate.

24. The method of claim 19 , wherein the substantially intact cell sheet comprises a plurality of cell types.

25. The method of claim 19 , wherein the substantially intact cell sheet comprises a plurality of cell layers.

26. The method of claim 19 , wherein the substantially intact cell sheet separated using fluidic shear stress is held intact by an endogenously produced scaffold.

27. The method of claim 20 , wherein the at least one cell-adherent patterned texture comprises a plurality of channels, at least

Assignees

• Upside Foods Inc

Inventors

• Leung Matthew
• Godbole Asha
• Engelmayr Jr George C
• Genovese Nicholas J
• Valeti Uma S
• Carswell Kathleen

Classifications

• C12N5/0625

  Epidermal cells, skin cells; Cells of the oral mucosa · CPC title

• C12N5/069

  Vascular Endothelial cells · CPC title

• C12N5/067

  Hepatocytes · CPC title

• C12N5/0655

  Chondrocytes; Cartilage · CPC title

• C12N5/0654

  Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth · CPC title