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CN111165738A - Edible bionic animal tissue - Google Patents

Edible bionic animal tissue Download PDF

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Publication number
CN111165738A
CN111165738A CN202010062023.0A CN202010062023A CN111165738A CN 111165738 A CN111165738 A CN 111165738A CN 202010062023 A CN202010062023 A CN 202010062023A CN 111165738 A CN111165738 A CN 111165738A
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China
Prior art keywords
edible
animal
biomimetic
animal tissue
derivatives
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CN202010062023.0A
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Chinese (zh)
Inventor
赵一麟
周媛媛
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Taiyang Yulin Xiamen Biomedical Co ltd
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Taiyang Yulin Xiamen Biomedical Co ltd
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Priority to CN202010062023.0A priority Critical patent/CN111165738A/en
Publication of CN111165738A publication Critical patent/CN111165738A/en
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/50—Poultry products, e.g. poultry sausages
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
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    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0653—Adipocytes; Adipose tissue
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0657—Cardiomyocytes; Heart cells
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    • C12N5/0602—Vertebrate cells
    • C12N5/067—Hepatocytes
    • C12N5/0671—Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/90—Substrates of biological origin, e.g. extracellular matrix, decellularised tissue

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Abstract

Disclosed is an edible biomimetic animal tissue comprising a natural material carrier and animal cells which grow on the natural material carrier by static or dynamic culture and secrete major extracellular matrix components including collagen, elastin, hyaluronic acid and glycosaminoglycan on the natural carrier to biomimetic living tissue. The edible bionic animal tissue is not simple artificial meat of plant source, is real animal tissue, is bionic living tissue, and can be used as meat for human beings to eat.

Description

Edible bionic animal tissue
Technical Field
The invention belongs to the technical field of food engineering, and particularly relates to edible bionic animal tissue.
Background
Human survival requires the ingestion of animal proteins, but in the process of obtaining animal proteins, humans acquire animal proteins by killing the animal. With the continued development of human civilization, various animal protective tissues are emerging that object and prevent the acquisition of animal proteins and related foods by killed animals. Some religions are convincing and are fundamentally resistant to eating all meat products obtained by damaging the life of animals, and some special religions consider certain animals to be magical and prohibited from eating.
At present, the scientific level is a trend to culture meat products in an in vitro tissue engineering mode and avoid killing animals through an artificial method.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide edible bionic animal tissue.
The technical scheme of the invention is as follows:
an edible biomimetic animal tissue comprising a natural material carrier and animal cells which grow on the natural material carrier by static or dynamic culture and secrete major extracellular matrix components including collagen, elastin, hyaluronic acid and glycosaminoglycan on the natural carrier to biomimetic living tissue;
the natural material carrier is made of at least one of animal polysaccharide, animal protein, plant polysaccharide, plant protein, algae polysaccharide and microorganism polysaccharide; or comprises fiber or protein main body structure of fructus Luffae, fructus Citri Grandis, bamboo shoot, radix et caulis Opuntiae Dillenii, fructus Solani Melongenae, herba Apii Graveolentis, edible fungi or edible algae, which is suitable for cell adhesion proliferation after removing impurities; or comprises fructus Luffae, fructus Citri Grandis, bamboo shoot, radix et caulis Opuntiae Dillenii, fructus Solani Melongenae, herba Apii Graveolentis, edible fungi or edible algae, pulverizing, removing impurities, and adding edible plasticizer to reform structure suitable for cell adhesion proliferation;
the animal origin polysaccharide comprises at least one of glycogen, bone glue, shellac, gelatin, sponge skeleton, chitin and its derivatives, and chitosan and its derivatives of lower animals;
the animal derived protein comprises egg derived protein, milk protein and silk fibroin;
the plant-derived polysaccharide comprises at least one of polysaccharide, monosaccharide, pectin and its derivatives, cyclodextrin and its derivatives, starch, modified starch, cellulose and modified cellulose;
the plant-derived protein includes protein extracted from beans, peanuts, corn, wheat, loofah, eggplant and celery;
the algae-derived polysaccharide comprises at least one of agar and derivatives thereof, carrageenan and derivatives thereof, and alginic acid and derivatives thereof;
the polysaccharide derived from microorganism comprises microbial cellulose, and the microbial cellulose comprises bacterial cellulose and derivatives thereof;
the animal cell is derived from edible mammals, birds or aquatic organisms. .
The edible mammals include pigs, rabbits, sheep, cattle, dogs and camels, the birds include chickens, ducks, geese, ostriches, quails and pigeons, the aquatic organisms include fish, shrimp and crab, and other lower marine aquatic organisms. The animal cell may be obtained by puncturing a living animal, or may be a cell from another source, or may be a cell commercially available.
In a preferred embodiment of the present invention, the edible biomimetic animal tissue is in the shape of a sheet, a block, a cuboid, an irregularity, and can be adjusted in size and thickness according to the culture equipment.
In a preferred embodiment of the present invention, the pectin is derived from at least one of lemon peel, grapefruit peel, aloe peel, orange peel and tangerine peel.
In a preferred embodiment of the present invention, the animal cell includes at least one of a muscle cell, an adipocyte, an epithelial cell, a fibroblast, an endothelial cell, a mesenchymal cell, a stem cell, a liver cell, and a cardiac muscle cell.
In a preferred embodiment of the invention, an edible nutritional supplement comprising at least one of fatty acids, carbohydrates, proteins, nutritional elements, trace elements and vitamins, and/or a food additive is also included.
in a preferred embodiment of the present invention, the natural material carrier is made from raw materials including chitosan, cellulose, β -cyclodextrin, gelatin, and glycerin.
In a preferred embodiment of the present invention, the natural material carrier is made from raw materials including bacterial cellulose, zein, and wheat gliadin.
In a preferred embodiment of the present invention, the natural material carrier is made from raw materials including pectin extracted from grapefruit peel, ascorbic acid, a calcium source, and chitosan.
Further preferably, the calcium source includes at least one of inorganic or organic substances containing calcium ions or calcium elements.
In a preferred embodiment of the present invention, the natural material carrier is a structure obtained by removing the skin from the shaddock peel, soaking in an alcohol solution of gradient concentration to remove impurities including easily-dissolved pigments, freeze-fixing at-80 ℃ and vacuum-drying to remove liquid components including moisture.
The invention has the beneficial effects that: the edible bionic animal tissue is not artificial meat simply from plant sources, is real animal protein, is bionic living tissue, and can be used as meat for human beings to eat.
Drawings
FIG. 1 is a photograph showing one of the edible biomimetic animal tissues prepared in example 2 of the present invention.
FIG. 2 is a second photograph of the edible biomimetic animal tissue prepared in example 2 of the present invention.
FIG. 3 shows the results of the cytotoxicity test of the natural material carrier in example 1 of the present invention
FIG. 4 is a view showing the configuration of a carrier of natural material according to example 2 of the present invention implanted in a rat subcutaneously for half a year.
FIG. 5 is a graph showing the contents of the components of the edible biomimetic animal tissue prepared in example 1 of the present invention.
FIG. 6 is a graph comparing the nutrient contents of the edible biomimetic animal tissue prepared in example 4 of the present invention with other meat.
Detailed Description
The technical solution of the present invention will be further illustrated and described below with reference to the accompanying drawings by means of specific embodiments.
Example 1 edible biomimetic animal tissue with chitosan-gelatin as scaffold
dissolving chitosan, cellulose, gelatin and β cyclodextrin (mass ratio is 1: 8: 0.5) in dilute acetic acid with volume fraction of 2%, and stirring thoroughly to obtain clear and transparent chitosan solution with mass concentration of 20 g/L.
Measuring 200mL of the chitosan solution, adding 0.5g of glycerol, adjusting the pH value of the mixed solution to 4.0, carrying out vacuum defoamation under 0.1Mpa for 0.5h, carrying out tape casting in a polyethylene disc, placing the solution in a vacuum drying oven, drying at 50 ℃ for 4h, and removing to obtain the natural material carrier.
Cytotoxicity assays were performed using hepatic stellate cells from rats: soaking 0.1g of the above natural material carrier in 100mL/37 deg.C culture medium for 24h, filtering the leaching solution with 0.22 μm filter head, planting 5000 cells in each well of 96-well plate, repeating 3 groups, detecting CCK-8 in 1, 3, 5, 7, and 9 days, respectively, recording absorbance, and using normal complete culture medium as negative control, the result is shown in FIG. 3.
Muscle cells of pig (1X 10)7cells/cm2) Adhering to the above natural material carrier, and culturing in a Belck Dualcell Trol bioreactor for 20 days to obtain edible bionic animal tissue (shown in FIG. 5, which has extracellular matrix including collagen, elastin, hyaluronic acid and glycosaminoglycan) with chitosan-gelatin as natural material carrier.
Example 2 edible biomimetic animal tissue with plant protein-bacterial cellulose as scaffold
Bacterial Cellulose (BC) is a generic term for cellulose synthesized by any of microorganisms belonging to the genera Acetobacter (Acetobacter), Agrobacterium (Agrobacterium), Rhizobium (Rhizobium), Sarcina (Sarcina), and the like under different conditions. The bacterial cellulose generated by the fermentation of the bacterial strain is subjected to purification treatment, cutting and freeze drying to obtain the bacterial cellulose.
(1) Adding zein and wheat gliadin into the ethanol solution for dissolving;
(2) adding bacterial cellulose into distilled water for dissolving;
(3) mixing zein, wheat prolamin solution and bacterial cellulose solution, adding glycerol, and performing crosslinking reaction for 1h (the mass ratio of zein to wheat prolamin to bacterial cellulose to glycerol is 1: 1.2: 0.5);
(4) pouring the solution on a smooth mould, and carrying out vacuum drying to obtain the natural material carrier;
(5) the muscle cells of sheep were adhered to a natural material carrier, and cultured in a Belck Dual CellTrol bioreactor for 20 days to obtain an edible biomimetic animal tissue (having an extracellular matrix including collagen, elastin, hyaluronic acid, and glycosaminoglycan) in which the plant protein-bacterial cellulose as a main scaffold as shown in FIGS. 1 and 2.
As sheep are large animals and the experiment is relatively complex, the histocompatibility experiment of the material adopts a rat to replace the experiment, in order to prevent rejection reaction, the cells of the sheep are removed and transplanted under the skin of the rat, and the observation is opened after one month. Biocompatibility was good, rats did not have any discomfort, growth was good, and the material was found to be obtained 1 month after transplantation, as shown in figure 4, the natural material carrier had been completely absorbed, indicating good material absorption.
Example 3 edible biomimetic animal tissue with grapefruit peel extract-chitosan-soy protein as scaffold
The pomelo peel is rich in protein, organic acid, vitamins and calcium, etc., and the main component of the pomelo peel extract is pectin.
Firstly, extracting a fat substitute containing active ingredients of the shaddock peel from the fresh shaddock peel, cutting the fresh shaddock peel into particles, putting the particles into water, heating the particles to 90 ℃, preserving the heat for 45min for enzyme deactivation, repeatedly pressing juice until washing liquor is colorless, and then pressing the juice to obtain pretreated shaddock peel; adding pretreated shaddock peel into 3% tartaric acid aqueous solution according to a feed-liquid ratio of 1: 10, heating to 60 ℃, and performing ultrasonic treatment for 20 min; filtering, cooling, and concentrating the filtrate to 20% of its volume to obtain pectin concentrate; preparing an ascorbic acid and 60 ℃ into an aqueous solution with the mass fraction of 60%, adding a calcium source, wherein the mass ratio of the ascorbic acid to the calcium source is 1: 0.4-0.6, and carrying out ultrasonic reaction for 5min to obtain a calcium ascorbate solution; the calcium source is at least one of calcium carbonate, calcium oxide and calcium hydroxide; adding ethanol water solution into calcium ascorbate solution according to 4-8 times volume of pectin concentrated solution, stirring thoroughly to disperse calcium ascorbate uniformly, adding into the obtained pectin concentrated solution, standing for 16-24h to form gel, which is fat substitute containing pericarpium Citri Grandis active ingredient.
Adjusting the soy protein isolate solution to an isoelectric point, centrifuging to remove albumin, adding water with the same volume, and adjusting the pH to 9.0; dissolving chitosan in 2% acetic acid solution; adjusting pH to 4.2-4.8, and adding the fat substitute containing active components of pericarpium Citri Grandis; mixing the three components, and freeze-drying and forming to obtain the natural material carrier.
Attaching bovine muscle cells to the natural material carrier, and culturing in Belck Dual CellTrol bioreactor for 20 days to obtain edible bionic animal tissue (with extracellular matrix including collagen, elastin, hyaluronic acid and glycosaminoglycan) with pericarpium Citri Grandis extract-chitosan-soybean protein as scaffold.
Example 4 edible biomimetic animal tissue with shaddock peel as structural scaffold after impurity removal
Peeling pericarpium Citri Grandis, cutting into small pieces of 10 × 10 × 1(cm), soaking in ethanol (100% ethanol or 70% ethanol) of different concentrations to remove easily-dissolved pigment and other impurities, freezing and fixing at-80 deg.C, vacuum drying, and removing water and other liquid components. The shaddock peel after impurity removal and drying is hard in texture, but strong in water absorption performance and can absorbThe water content is about 5 times of the volume of the water. Soaking dried pericarpium Citri Grandis in cell complete culture medium, incubating for 24 hr, removing excess culture medium, adding polylysine (10 μ g/ml) in CO2Incubating at 37 deg.C for 12-24 hr, drying, and planting porcine carotid endothelial cells (amount of planted cells: the above-mentioned material with size of 2 × 10)7cells; the amount of medium is generally the water uptake of the material, about 4-5 ml; after the cells are planted for 4-6h, supplementing a culture medium in the culture medium after the cells adhere to the wall, and keeping the material in a humid environment; observe changes in cell culture medium, change every 2-4 days). Culturing for 3-4 weeks to obtain edible bionic animal tissue.
The main components of the edible bionic animal tissue are detected and compared with beef, lean pork and mutton, and the result is shown in figure 6.
And (3) measuring the moisture content: reference is made to GB 5009.3-2010 national food safety Standard determination of moisture; protein content determination: reference is made to GB 5009.5-2010 "determination of proteins in food safety national standard foods"; fat content determination: refer to GB/T9695.7-2008 'determination of total fat content of meat and meat products'. In addition, except for the shaddock peel used in the embodiment, eggplant, cactus, towel gourd, bamboo shoot, mushroom and the like can be made into edible bionic animal tissues in the same or similar mode.
The above description is only a preferred embodiment of the present invention, and therefore should not be taken as limiting the scope of the invention, which is defined by the appended claims.

Claims (9)

1. An edible biomimetic animal tissue, characterized in that: comprises a natural material carrier and animal cells which grow on the natural material carrier by static or dynamic culture and secrete major extracellular matrix components including collagen, elastin, hyaluronic acid and glycosaminoglycan on the natural carrier to simulate living tissue;
the natural material carrier is made of at least one of animal polysaccharide, animal protein, plant polysaccharide, plant protein, algae polysaccharide and microorganism polysaccharide; or comprises fiber or protein main body structure of fructus Luffae, fructus Citri Grandis, bamboo shoot, radix et caulis Opuntiae Dillenii, fructus Solani Melongenae, herba Apii Graveolentis, edible fungi or edible algae, which is suitable for cell adhesion proliferation after removing impurities; or comprises fructus Luffae, fructus Citri Grandis, bamboo shoot, radix et caulis Opuntiae Dillenii, fructus Solani Melongenae, herba Apii Graveolentis, edible fungi or edible algae, pulverizing, removing impurities, and adding edible plasticizer to reform structure suitable for cell adhesion proliferation;
the animal origin polysaccharide comprises at least one of glycogen, bone glue, shellac, gelatin, sponge skeleton, chitin and its derivatives, and chitosan and its derivatives of lower animals;
the animal derived protein comprises egg derived protein, milk protein and silk fibroin;
the plant-derived polysaccharide comprises at least one of polysaccharide, monosaccharide, pectin and its derivatives, cyclodextrin and its derivatives, starch, modified starch, cellulose and modified cellulose;
the plant-derived protein includes protein extracted from beans, peanuts, corn, wheat, loofah, eggplant and celery;
the algae-derived polysaccharide comprises at least one of agar and derivatives thereof, carrageenan and derivatives thereof, and alginic acid and derivatives thereof;
the polysaccharide derived from microorganism comprises microbial cellulose, and the microbial cellulose comprises bacterial cellulose and derivatives thereof; the animal cell is derived from edible mammals, birds or aquatic organisms.
2. An edible biomimetic animal tissue as recited in claim 1, wherein: the pectin and its derivatives are derived from at least one of lemon, grapefruit peel, mandarin orange peel, orange peel and tangerine peel.
3. An edible biomimetic animal tissue as recited in claim 1, wherein: the animal cells include at least one of muscle cells, adipocytes, epithelial cells, fibroblasts, endothelial cells, mesenchymal cells, stem cells, liver cells, and cardiac muscle cells.
4. An edible biomimetic animal tissue as recited in claim 1, wherein: also included are dietary nutritional supplements and/or food additives.
5. an edible biomimetic animal tissue according to any of claims 1-4, wherein the natural material carrier is made from materials including chitosan, β -cyclodextrin, cellulose, gelatin, and glycerin.
6. An edible biomimetic animal tissue according to any of claims 1-4, wherein: the natural material carrier is made of raw materials including bacterial cellulose, zein and wheat gliadin.
7. An edible biomimetic animal tissue according to any of claims 1-4, wherein: the natural material carrier is prepared from raw materials including pectin extracted from pericarpium Citri Grandis, ascorbic acid, calcium source and chitosan.
8. An edible biomimetic animal tissue as recited in claim 7, wherein: the calcium source includes at least one of calcium carbonate, calcium oxide, and calcium hydroxide.
9. An edible biomimetic animal tissue according to any of claims 1-4, wherein: the natural material carrier is a structure obtained by removing peel of shaddock peel, soaking in alcohol solution with gradient concentration to remove impurities including pigment easy to dissolve out, freezing and fixing at-80 ℃, and vacuum drying to remove liquid components including water.
CN202010062023.0A 2020-01-19 2020-01-19 Edible bionic animal tissue Pending CN111165738A (en)

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Cited By (7)

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CN113208059A (en) * 2020-12-14 2021-08-06 西北农林科技大学 Method for manufacturing edible pectin chitosan collagen 3D scaffold for cell culture meat
CN114438615A (en) * 2022-02-28 2022-05-06 上海食未生物科技有限公司 Large-scale production method of soybean protein fiber scaffold for cell culture meat
CN114438014A (en) * 2022-01-26 2022-05-06 上海食未生物科技有限公司 An edible chitosan glutenin biomimetic oriented cell culture meat bioscaffold
CN114752067A (en) * 2022-05-18 2022-07-15 上海食未生物科技有限公司 Modified bacterial cellulose beneficial to cell culture and application thereof
CN115697079A (en) * 2020-06-12 2023-02-03 生物技术食品有限公司 Edible and sterilizable porous 3D scaffold and its use
CN117500385A (en) * 2021-06-11 2024-02-02 阿普赛德食品公司 Cell-based edible meat products including dried cell powder and methods of making the same
CN117586949A (en) * 2023-11-24 2024-02-23 晨熙新创生物科技(镇江)有限公司 Plant fiber scaffold and preparation method and application thereof

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115697079A (en) * 2020-06-12 2023-02-03 生物技术食品有限公司 Edible and sterilizable porous 3D scaffold and its use
CN113208059A (en) * 2020-12-14 2021-08-06 西北农林科技大学 Method for manufacturing edible pectin chitosan collagen 3D scaffold for cell culture meat
CN117500385A (en) * 2021-06-11 2024-02-02 阿普赛德食品公司 Cell-based edible meat products including dried cell powder and methods of making the same
CN114438014A (en) * 2022-01-26 2022-05-06 上海食未生物科技有限公司 An edible chitosan glutenin biomimetic oriented cell culture meat bioscaffold
CN114438615A (en) * 2022-02-28 2022-05-06 上海食未生物科技有限公司 Large-scale production method of soybean protein fiber scaffold for cell culture meat
CN114752067A (en) * 2022-05-18 2022-07-15 上海食未生物科技有限公司 Modified bacterial cellulose beneficial to cell culture and application thereof
CN114752067B (en) * 2022-05-18 2024-02-06 上海食未生物科技有限公司 Modified bacterial cellulose beneficial to cell culture and application thereof
CN117586949A (en) * 2023-11-24 2024-02-23 晨熙新创生物科技(镇江)有限公司 Plant fiber scaffold and preparation method and application thereof

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