Nutrient media for the production of slaughter-free meat
Provided herein are nutrient media formulations and engineered growth factors, and methods thereof, useful for the production of slaughter-free meat.
1. A method for formulating an edible nutrient medium for the production of slaughter-free meat for human consumption, comprising:
(a) providing a plurality of ingredients including
(i) one or more of cyanocobalamin, an edible anti-foaming agent, and ferric ammonium citrate; and
(ii) one or more of IGF-1, LR3-IGF1, FGF-1, FGF-2, PDGF, CTGF, EGF, TGFB, BMP, HGF, transferrin, insulin, WNT proteins, interleukins, albumin, VEGF, and homologs thereof; and
(b) mixing the plurality of ingredients in water to form the edible nutrient medium; wherein the nutrient medium does not comprise cobalt chloride, ammonium molybdate, ammonium metavanadate, sodium metavanadate, rubidium chloride, and strontium chloride hexahydrate; thereby forming an edible nutrient medium for culturing an animal cell line suitable for the production of slaughter-free meat.
2. The method of claim 1 , wherein the nutrient medium further comprises 8 or more amino acids selected from those below and within concentration ranges shown below:
Lowest
Highest
Concentration
Concentration
Amino Acids
(g/L)
(g/L)
L-alanine
0.0001
0.5
L-arginine
0.01
2.5
L-arginine hydrochloride
0.01
1
L-asparagine anhydrous
0.1
1.5
L-asparagine monohydrate
0.01
0.9
L-cysteine hydrochloride
0.001
0.5
L-cystine dihydrochloride
0.005
1.5
L-glutamic acid
0.01
1.5
L-glutamic acid monosodium salt
0.01
1.5
monohydrate
L-glutamine
0.14
1.5
L-glycine
0.01
0.5
L-histidine free base
0.01
0.5
L-histidine hydrochloride
0.01
0.5
hydroxy-L-proline
0.01
0.25
L-isoleucine
0.1
0.75
L-leucine
0.01
0.75
L-lysine
0.01
1
L-methionine
0.05
0.5
L-phenylalanine
0.05
0.5
L-proline
0.1
1
L-serine
0.1
1
L-threonine
0.05
0.5
L-tryptophan
0.01
0.5
L-tyrosinedisodium saltdihydrate
0.01
0.5
L-valine
0.1
0.75
L-ornithine
0.001
0.05
L-aspartic acid
0.1
0.8
L-alpha-aminobutyric acid
0.001
0.025
Gaba
0.01
0.2
l-carnosine
0.01
1.
3. The method of claim 1 , further comprising providing plant derived oils or yeast hydrolysates to the nutrient medium, wherein the nutrient medium does not comprise DL-alpha lipoic acid.
4. The method of claim 1 , wherein the nutrient medium does not comprise: one or more category 3 ingredients selected from: bovine transferrin, chicken serum, and bovine serum; and
the plurality of ingredients includes d-glucose, folic acid, dl-alpha-tocopherol acetate, vitamin d2, d-alpha-tocopherol, d-biotin, myo-lnositol, niacinamide, pyri doxine-hcl, pyri doxal-hcl, riboflavin, thiamine-hcl, vitamin b12, calcium d-pantothenate, choline chloride, ascorbic acid, sodium chloride, calcium chloride, manganese sulfate H2O, potassium chloride, magnesium chloride, copper sulfate-pentahydrate, sodium metasilicate-9H2O, zinc sulfate-7H2O, magnesium sulfate, sodium phosphate monobasic H2O, sodium phosphate dibasic, 1-alanine, 1-arginine-hcl, 1-asparagine-H2O, l-aspartic acid, l-cysteine hcl H2O, l-cystine 2hcl, l-glutamic acid monosodium H2O, l-glutamine, glycine, l-histidine-hcl-H2O, l-isoleucine, l-leucine, l-lysine-hcl, l-methionine, l-phenylalanine, l-proline, l-serine, l-threonine, l-tryptophan, l-tyrosine-2na-2H2O, l-valine, succinic acid, sodium succinate-2na 6H2O, sodium selenite (anhy), ferrous sulfate-7H2O, ferric citrate, ferric ammonium citrate, linoleic acid, monoethanola mine, tween 80, l-glutathione, and hypep 2 yeast extract.
5. The method of claim 1 , wherein the nutrient medium does not comprise: rubidium chloride, strontium chloride hexahydrate, hypoxanthine, putrescine, thymidine, and DL-alpha-lipoic acid.
6. The method of claim 1 , wherein the nutrient medium does not comprise: chicken serum and bovine serum.
7. The method of claim 1 , wherein the nutrient medium does not comprise any animal-derived component nor any animal-derived serum.
8. The method of claim 1 , wherein the plurality of ingredients includes one or more of glycolysis intermediates, iron carriers, ferric maltol, ferrous gluconate, shear protectants, polyethylene glycol, methylcellulose, simethicone, food-grade plant oils, and hydrolysates.
9. The method of claim 1 , wherein the plurality of ingredients includes one or more of an adhesion protein selected from Vitronectin; Collagen alpha-3(IV); Collagen, type IV, alpha I; TPA: fibronectin; and Fibronectin, partial.
10. The method of claim 1 , wherein:
(a) the nutrient medium does not comprise poloxamer;
(b) the plurality of ingredients includes methylcellulose in an amount to provide shear protection; and
(c) the plurality of ingredients includes cyanocobalamin to provide an essential cofactor.
11. The method of claim 1 , wherein the plurality of ingredients includes glycolysis intermediates, shear protectants, polyethylene glycol, methylcellulose, food-grade plant oils, hydrolysates, and one or more of iron carriers, ferric maltol, ferrous gluconate, and an adhesion protein selected from Vitronectin; Collagen alpha-3(IV); Collagen, type IV, alpha I; TPA: fibronectin; and Fibronectin, partial.
12. A method for formulating an edible nutrient medium for the production of cultured meat for human consumption, comprising:
(a) providing a plurality of ingredients including one or more of cyanocobalamin, an edible anti-foaming agent, and ferric ammonium citrate; and
(b) mixing the plurality of ingredients in water to form the edible nutrient medium; wherein the nutrient medium does not comprise cobalt chloride, ammonium molybdate, ammonium metavanadate, sodium metavanadate, rubidium chloride, strontium chloride hexahydrate, animal-derived components, and animal-derived serums.
13. The method of claim 12 , wherein the plurality of ingredients includes two or more of cyanocobalamin, an edible anti-foaming agent, and ferric ammonium citrate.
14. The method of claim 12 , wherein the plurality of ingredients includes methylcellulose.
15. The method of claim 12 , wherein the nutrient medium further comprises 8 or more amino acids selected from those below and within concentration ranges shown below:
Lowest
Highest
Concentration
Concentration
Amino Acids
(g/L)
(g/L)
L-alanine
0.0001
0.5
L-arginine
0.01
2.5
L-arginine hydrochloride
0.01
1
L-asparagine anhydrous
0.1
1.5
L-asparagine monohydrate
0.01
0.9
L-cysteine hydrochloride
0.001
0.5
L-cystine dihydrochloride
0.005
1.5
L-glutamic acid
0.01
1.5
L-glutamic acid monosodium salt
0.01
1.5
monohydrate
L-glutamine
0.14
1.5
L-glycine
0.01
0.5
L-histidine free base
0.01
0.5
L-histidine hydrochloride
0.01
0.5
hydroxy-L-proline
0.01
0.25
L-isoleucine
0.1
0.75
L-leucine
0.01
0.75
L-lysine
0.01
1
L-methionine
0.05
0.5
L-phenylalanine
0.05
0.5
L-proline
0.1
1
L-serine
0.1
1
L-threonine
0.05
0.5
L-tryptophan
0.01
0.5
L-tyrosinedisodium saltdihydrate
0.01
0.5
L-valine
0.1
0.75
L-ornithine
0.001
0.05
L-aspartic acid
0.1
0.8
L-alpha-aminobutyric acid
0.001
0.025
Gaba
0.01
0.2
l-carnosine
0.01
1.
16. A method for formulating an edible nutrient medium for the production of cultured meat for human consumption, comprising:
(a) providing a plurality of ingredients including:
(i) one or more of cyanocobalamin, an edible anti-foaming agent, and ferric ammonium citrate; and
(ii) a plurality of amino acids within concentration ranges shown below:
Lowest
Highest
Concentration
Concentration
Amino Acids
(g/L)
(g/L)
L-alanine
0.0001
0.5
L-arginine
0.01
2.5
L-arginine hydrochloride
0.01
1
L-asparagine anhydrous
0.1
1.5
L-asparagine monohydrate
0.01
0.9
L-cysteine hydrochloride
0.001
0.5
L-cystine dihydrochloride
0.005
1.5
L-glutamic acid
0.01
1.5
L-glutamic acid monosodium salt
0.01
1.5
monohydrate
L-glutamine
0.14
1.5
L-glycine
0.01
0.5
L-histidine free base
0.01
0.5
L-histidine hydrochloride
0.01
0.5
hydroxy-L-proline
0.01
0.25
L-isoleucine
0.1
0.75
L-leucine
0.01
0.75
L-lysine
0.01
1
L-methionine
0.05
0.5
L-phenylalanine
0.05
0.5
L-proline
0.1
1
L-serine
0.1
1
L-threonine
0.05
0.5
L-tryptophan
0.01
0.5
L-tyrosinedisodium saltdihydrate
0.01
0.5
L-valine
0.1
0.75
L-ornithine
0.001
0.05
L-aspartic acid
0.1
0.8
L-alpha-aminobutyric acid
0.001
0.025
Gaba
0.01
0.2
l-carnosine
0.01
1
and
(b) mixing the plurality of ingredients in water to form the edible nutrient medium; wherein the nutrient medium does not comprise cobalt chloride, ammonium molybdate, ammonium metavanadate, sodium metavanadate, rubidium chloride, and strontium chloride hexahydrate.
17. The method of claim 16 , wherein the nutrient medium does not comprise animal-derived components or animal-derived serums.
18. The method of claim 16 , wherein the nutrient medium does not comprise: rubidium chloride, strontium chloride hexahydrate, hypoxanthine, putrescine, thymidine, and DL-alpha-lipoic acid.
19. The method of claim 16 , wherein the plurality of ingredients includes methylcellulose.