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JP-7855209-B2 - Cultured meat composition

JP7855209B2JP 7855209 B2JP7855209 B2JP 7855209B2JP-7855209-B2

Inventors

  • ベン－アリー，トム
  • レーベンバーグ，シュラミット

Assignees

  • アレフ ファームス リミテッド

Dates

Publication Date
20260508
Application Date
20180715
Priority Date
20170715

Claims (20)

  1. A method for producing an edible composition, a. An edible three-dimensional porous scaffold containing protein, wherein the protein is selected from the group consisting of textured proteins and non-textured proteins, and (i) Myoblasts or their precursor cells, (ii) at least one extracellular matrix (ECM) secreting cell selected from the group consisting of fibroblasts, smooth muscle cells, stromal cells, pericytes, and their precursor cells, and (iii) Incubating multiple non-human cell types, including endothelial cells or their precursor cells, The ratio of myoblasts or their progenitor cells , ECM-secreting cells, and endothelial cells or their progenitor cells is 2:1:1 , and incubation is performed. b. Inducing myoblasts or their precursor cells to differentiate into myotubes, A method comprising, by means of, inducing the production of the aforementioned edible composition.
  2. The method according to claim 1, wherein the myoblast progenitor cells are satellite cells.
  3. The method according to claim 1, characterized in that the myoblasts or their precursor cells and the endothelial cells or their precursor cells are incubated for a period necessary for the myoblasts or their precursor cells to achieve a coverage rate of at least 30% of the edible three-dimensional porous scaffold, and then the differentiation of the myoblasts or their precursor cells into myotubes is induced.
  4. The method according to any one of claims 1 to 3, wherein the endothelial cells are skeletal microvascular endothelial cells.
  5. The method according to any one of claims 1 to 4, wherein the plurality of cell types include satellite cells, fibroblasts, and endothelial cells.
  6. The method according to any one of claims 1 to 4, wherein the plurality of cell types include satellite cells, smooth muscle cells, and endothelial cells.
  7. The method according to claim 1, wherein the protein is a textured protein.
  8. The method according to claim 7 , wherein the textured protein is textured soy protein.
  9. The method according to claim 7 or 8 , wherein the three-dimensional porous scaffold includes pores having an average diameter in the range of 20 to 1,000 micrometers.
  10. The method according to any one of claims 1 to 9 , wherein the plurality of cells are derived from livestock mammals.
  11. The method according to claim 10, wherein the livestock mammal is a cattle.
  12. The method according to any one of claims 1 to 11, wherein the myoblasts or their progenitor cells and the three-dimensional porous scaffold are incubated in 10 mg of the three-dimensional porous scaffold in a ratio ranging from 10³ to 10⁷ cells .
  13. The method according to any one of claims 1 to 12, wherein the three-dimensional porous scaffold further comprises an extracellular matrix.
  14. a. An edible three-dimensional porous scaffold containing protein, wherein the protein is selected from the group consisting of textured proteins and non-textured proteins, b. The three-dimensional porous scaffold contains myotubes with 10,000 to 250,000 myotube nuclei per 1 mm³, c. Multiple cells including multiple cell types, (i) Myoblasts or their precursor cells, (ii) at least one extracellular matrix (ECM) secreting cell selected from the group consisting of fibroblasts, smooth muscle cells, stromal cells, pericytes, and their precursor cells, and (iii) A plurality of cells comprising a plurality of cell types , which constitute less than 15% of the plurality of cells, and which are endothelial cells or their precursor cells, The aforementioned plurality of cell types are cultured in a ratio of (i):(ii):(iii) of 2:1:1 in this edible composition.
  15. The composition according to claim 14, wherein the myoblast progenitor cells are satellite cells.
  16. The composition according to claim 14 or 15, wherein the plurality of cell types include satellite cells, fibroblasts, and endothelial cells.
  17. The composition according to claim 14 or 15, wherein the plurality of cell types include satellite cells, smooth muscle cells, and endothelial cells.
  18. The composition according to claim 15, wherein the endothelial cells are skeletal microvascular endothelial cells.
  19. The composition according to any one of claims 14 to 18 , wherein the protein is a textured protein.
  20. The composition according to claim 19 , wherein the textured protein is textured soy protein.

Description

Related Application This application claims priority to U.S. Provisional Patent Application No. 62/532,998, filed on 15 July 2017, the contents of which are incorporated herein by reference in their entirety. This invention relates, in particular, to a cultured meat composition and a method for producing the same. Cultured meat, also known as artificial meat or clean meat, is produced from cell cultures using tissue engineering technology and is a significant alternative to conventional meat production that uses live animals. Over the past decade, the concept has gained increasing attention from the public, influential media, investors, and the scientific community, particularly after the production of the first cultured beef burger. Producing food using animals is considered inefficient because animals consume vast amounts of food throughout their lives. In this production, 80-90% of calories are wasted in animal metabolism and the production of non-edible tissues. While meat has the greatest environmental impact when comparing various industries, generally speaking, all animal-based products have a larger environmental footprint compared to plant-based products in terms of soil and water demand, as well as greenhouse gas (GHG) emissions. According to a report by the United Nations Food and Agriculture Organization, the livestock sector accounts for 18% of GHG emissions, uses 30% of the Earth's landform or 70% of arable land, and 8% of the world's freshwater. Furthermore, global meat demand is projected to double by 2050, meaning that conventional meat production systems are unsustainable. Compared to several meat sources, cultured meat is estimated to reduce energy use by 7–45%, GHG emissions by 78–96%, land use by 99%, and water use by 82–96%. Intensive factory farming and inadequate animal welfare conditions contribute to the spread of foodborne diseases such as swine and avian influenza, as well as E. coli, Salmonella, and Campylobacter bacteria that can be found in meat. Producing meat in a sterile environment can help improve food safety. Furthermore, 70% of all antibiotics used in the United States are given to livestock as food additives, which promote the selection of antimicrobial-resistant strains and increase the likelihood of multidrug-resistant bacteria. The overuse of antibiotics is a major cause of the emergence of antibiotic-resistant bacteria, resulting in an economic burden of $55 billion annually in the United States alone, 2 million infections, 250,000 hospitalizations, and at least 23,000 deaths. In recent years, bacteria resistant to colistin, a last resort antibiotic, have emerged in pig farms in China. Current cultured meat technology focuses on satellite cell culture. Cells are grown, separated, differentiated, and harvested on microcarriers in two-dimensional (2D) flasks or suspensions. However, tissue is not composed solely of cells. The extracellular matrix (ECM), which consists of macromolecules such as glycoproteins and oligosaccharides and confers biochemical and biomechanical properties to tissue, constitutes the majority of tissue. The ECM regulates cell behavior and influences its composition. Therefore, ECM-producing cells are essential for cultured meat, and processes such as cell harvesting should be avoided. Furthermore, three-dimensional (3D) cell culture mimics the natural cellular environment and is crucial for precise cell behavior that influences the biochemical contents of cells. Figures 1A-1C. Photographs showing commercially available TSPs. (A) shows large, medium, and small chunk products of TSP. (B) and (C) show two other commercially available TSP flake products annotated as TVP (left) and Arcon (right).Figures 2A and 2B. Photographs showing the preparation of the TSP scaffold (A) and the TSP scaffold containing cells (B).Figures 3A and 3B. SEM images at 100x magnification of a large TSP (A) and a moderate TSP (B).Figures 4A and 4B. SEM images at 2x10⁵ magnification of a large TSP (A) and a moderate TSP (B).Figures 5A-5C. Confocal microscope images of TSP scaffolds incorporating fibroblasts (red) and endothelial cells (green) on days 8 (A), 18 (B), and 21 (C) after fibroblast seeding.These are confocal microscope images taken 14 days after seeding 200,000 fibroblasts (red) into three different scaffold samples (Samples 1-3) obtained from different sources, indicated as large, medium, small, and 90° to medium. The scaffold indicated as 90° to medium was cut from the pore side of the TSP scaffold prepared using the TSP scaffold procedure. Scale bar = 1 mm.These are confocal microscope images of myoblasts cultured on a TSP scaffold for 14 days (blue - DAPI, green - desmin).These are confocal microscope images of bovine skeletal muscle cells cultured on a TSP scaffold for 14 days (blue - DAPI, green - phalloidin).Figures 9A–9E. Micrographs of bovine aortic smooth muscle cells (BAOSMCs) (8 passages) seeded in different media. (A) Basal medium, (B) Commercial med