Cultivated meat scaffold, method for producing same, and cultivated meat containing same

Abstract

The present invention relates to a cultivated meat scaffold, a method for producing same, and a cultivated meat containing same. The cultivated meat scaffold according to the present invention comprises nanocellulose and a polymer hydrogel.

Description

Cultured meat scaffold, method for producing the same, and cultured meat comprising the same

[0001] The present invention relates to a cultured meat scaffold, a method for producing the same, and cultured meat comprising the same.

[0002] Cultured meat, produced from cell cultures using tissue engineering technology, is a promising alternative to traditional meat production using live animals. Meat produced from animals, particularly cattle, requires extensive resources—water, grain, land, and energy—and generates significant greenhouse gas emissions. In contrast, cultured meat uses less energy and resources and can reduce greenhouse gas emissions. Furthermore, intensive factory farming and poor animal welfare conditions for meat production can contribute to foodborne illnesses, with many farmed animals receiving growth hormones and antibiotics, which are ultimately passed on to consumers. Conversely, producing meat in a sterile environment can help improve food safety. Meat produced under rigorous regulations and controlled conditions is less susceptible to contaminant transmission and is safer for consumption. Furthermore, controlling protein content, fat content, and composition, as well as essential trace elements and vitamins, such as iron and zinc, can enhance the nutritional value of meat and make it healthier. However, it is a difficult task to increase the safety of meat products while improving their visual, nutritional, taste, and aroma characteristics.

[0003] Typically, cell growth occurs through a series of processes, including cell attachment, proliferation, differentiation, and organization. Cell organization is essential for the production of cultured meat, and this organization allows for texture and nutrition similar to real meat. However, cell organization is difficult when cultured in conventional two-dimensional cell culture dishes. In cultured meat production, a scaffold on which cells can attach and grow temporarily or permanently influences cell growth and is an essential element that enables three-dimensional cell organization. However, materials used as existing cultured meat scaffolds have limitations, such as low mechanical properties that prevent them from maintaining their shape in a cell culture environment. To address this issue, chemical cross-linking agents can be added to enhance the scaffold's properties, but this can lead to cytotoxicity. Therefore, there is a pressing need to develop a scaffold that can achieve a texture similar to real meat without chemical cross-linking agents, minimize cytotoxicity and the burden of ingestion, and facilitate easy and rapid cell culture.

[0004] The background technology described above is something that the inventor possessed or acquired in the process of deriving the disclosure of the present application, and cannot necessarily be said to be a publicly known technology disclosed to the general public prior to the present application.

[0005] In order to solve the above-described problem, the present invention aims to provide a cultured meat scaffold having excellent physical properties and excellent cell culture activity, a method for producing the same, and cultured meat comprising the same.

[0006] However, the problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by a person having ordinary skill in the relevant technical field from the description below.

[0007] The cultured meat scaffold according to the present invention comprises nanocellulose; and polymer hydrogel.

[0008] In one embodiment, the nanocellulose may be in the form of a powder, suspension or hydrogel.

[0009] According to one embodiment, the nanocellulose may have a diameter of 1 nm to 10 μm.

[0010] In one embodiment, the nanocellulose may be a hydrogel having a viscosity of 12,000 cP or more and a gelation time of 1 second to 30 seconds.

[0011] In one embodiment, the nanocellulose may have an endotoxin content of 5 EU / mL or less.

[0012] According to one embodiment, the polymer hydrogel may include at least one selected from the group consisting of collagen, chitosan, fibrin, agar, starch, alginate, hyaluronic acid, gelatin, dextran, elastin, carrageenan, polyethylene glycol, polyacrylic acid, polyvinyl alcohol, polyvinylpyrrolidone, polylactic acid, and polyglycolic acid.

[0013] In one embodiment, the polymer hydrogel may be in the form of beads or in bulk form.

[0014] According to one embodiment, the weight ratio of the nanocellulose and the polymer hydrogel may be 1:4 to 3:2.

[0015] In one embodiment, the scaffold may have a pore diameter of 5 μm to 500 μm, a porosity of 70% to 95%, and a pore uniformity of 10% to 40%.

[0016] In one embodiment, the scaffold has a density of 1 g / cm 3 10 g / cm 3 , and the strength may be 0.1 MPa to 20 MPa and the endotoxin may be 5 EU / mL or less.

[0017] A method for manufacturing a cultured meat scaffold according to the present invention comprises the steps of: preparing nanocellulose; mixing the nanocellulose with a polymer hydrogel; and freeze-drying.

[0018] According to one embodiment, the step of preparing the nanocellulose may include a step of treating the cellulose at a temperature of 20° C. to 50° C. for 30 minutes to 3 hours; a washing step; and a nanofabricating step.

[0019] According to one embodiment, the freeze-drying step may be freeze-drying at a temperature of -50°C to -80°C for 48 to 72 hours.

[0020] The cultured meat according to the present invention is produced by culturing cells or tissues on a cultured meat scaffold according to the present invention or a cultured meat scaffold produced by a method for producing a cultured meat scaffold according to the present invention.

[0021] In one embodiment, the scaffold may be capable of culturing cells or tissues while maintaining a three-dimensional structure.

[0022] In one embodiment, the cell may comprise at least one selected from the group consisting of a stem cell, a bone cell, a muscle cell, an adipocyte, and a stem cell.

[0023] In one embodiment, the cell or tissue survival rate after culture may be 80% or more, and the culture activity may be 20% or more.

[0024] The present invention can provide a cultured meat scaffold having excellent physical properties and excellent cell culture activity, a method for producing the same, and cultured meat comprising the same.

[0025] Specifically, the cultured meat scaffold according to the present invention includes nanocellulose and a polymer hydrogel, thereby being free of cytotoxicity and improving the physical properties of the scaffold by controlling the pore size and density, thereby exhibiting excellent culture activity of cells or tissues during cultured meat production, thereby enabling cultured meat to be provided with a texture similar to meat easily and quickly.

[0026] Figures 1a and 1b are images of cultured meat according to examples and comparative examples of the present invention.

[0027] Figures 2a and 2b are images of cultured meat scaffolds according to examples and comparative examples of the present invention.

[0028] Figures 3a and 3b are SEM images of cultured meat scaffolds according to examples and comparative examples of the present invention.

[0029] Figures 4a and 4b are images confirming cell culture in a cultured meat scaffold according to an example and comparative example of the present invention.

[0030] Figure 5 is a graph showing cell survival rate according to an embodiment of the present invention.

[0031] Hereinafter, embodiments are described in detail with reference to the attached drawings. However, the embodiments may be modified in various ways, and the scope of the patent application is not limited or restricted by these embodiments. It should be understood that all modifications, equivalents, or alternatives to the embodiments are included within the scope of the patent application.

[0032] The terms used in the examples are for illustrative purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "comprise" or "have" are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0033] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0034] In addition, when describing with reference to the attached drawings, the same components will be given the same reference numerals regardless of the drawing numbers, and redundant descriptions thereof will be omitted. When describing an embodiment, if it is determined that a detailed description of a related known technology may unnecessarily obscure the gist of the embodiment, the detailed description thereof will be omitted. In addition, when describing a component of an embodiment, terms such as first, second, A, B, (a), (b), etc. may be used. These terms are only to distinguish the component from other components, and the nature, order, or sequence of the component is not limited by the terms. When a component is described as being "connected," "coupled," or "connected" to another component, it should be understood that the component may be directly connected or connected to the other component, but another component may also be "connected," "coupled," or "connected" between each component.

[0035] Components included in one embodiment and components with common functions will be described using the same names in other embodiments. Unless otherwise stated, the descriptions given in one embodiment can be applied to other embodiments, and detailed descriptions will be omitted to the extent of overlap.

[0036]

[0037] The cultured meat scaffold according to the present invention comprises nanocellulose; and polymer hydrogel.

[0038] The cultured meat scaffold according to the present invention provides a surface and support for attachment, maturation, differentiation, and proliferation of cells for producing cultured meat, and can complement the limitations of materials conventionally used as cultured meat scaffolds, such as having low mechanical properties to the extent that they cannot maintain their shape in a cell culture environment, or having low cell adhesion and thus difficulty in cell growth when produced using only a single material. That is, by including nanocellulose and polymer hydrogel, the pore size and density can be controlled, and accordingly, the physical properties of the scaffold and metabolism for nutrient supply and increased waste discharge are improved, thereby increasing culture activity and providing cultured meat with excellent texture.

[0039] The above nanocellulose has high miscibility and can be easily mixed with the polymer hydrogel to compensate for the low mechanical properties of the hydrogel.

[0040] The above cultured meat scaffold may further comprise nanochitin or a levan-catechol conjugate.

[0041]

[0042] In one embodiment, the nanocellulose may be in the form of a powder, suspension or hydrogel.

[0043] Nanocellulose according to the present invention is in the form of powder, suspension or hydrogel, and a suitable form of nanocellulose can be selected as needed.

[0044] Nanocellulose is a polymer material that is made by breaking down cellulose into nanosized pieces by one billionth of a meter. It is five times stronger than iron, five times lighter, flexible, environmentally friendly, and has antibacterial properties.

[0045]

[0046] According to one embodiment, the nanocellulose may have a diameter of 1 nm to 10 μm.

[0047] The above nanocellulose diameter may preferably be 1 nm to 1 μm; 1 nm to 500 nm; 1 nm to 100 nm; 1 nm to 50 nm; 50 nm to 10 μm; 50 nm to 1 μm; 50 nm to 500 nm; 50 nm to 100 nm; 100 nm to 10 μm; 100 nm to 1 μm; 100 nm to 500 nm; 500 nm to 10 μm; 500 nm to 1 μm; or 1 μm to 10 μm.

[0048]

[0049] If the diameter is less than the above range, there may be a problem in that the mechanical strength is reduced and it is difficult to maintain the structure, and if it exceeds the above range, there may be a problem in that the effect of the scaffold is reduced due to an increase in pores and a decrease in specific surface area.

[0050]

[0051] In one embodiment, the nanocellulose may be a hydrogel having a viscosity of 12,000 cP or more and a gelation time of 1 second to 30 seconds.

[0052] If the above nanocellulose is in hydrogel form, its viscosity may be 12,000 cP or higher. If the viscosity is below this range, scaffolds may not form properly. If the viscosity is above this range, it has the advantage of maintaining a stable structure under various environmental conditions.

[0053] The above nanocellulose may have a gelation time of preferably 1 to 20 seconds, and more preferably 1 to 10 seconds. Conventional nanocellulose has had problems such as not being gelled or requiring the addition of specific substances for gelation, but the nanocellulose according to the present invention does not require separate additives due to its short gelation time, can shorten the scaffold manufacturing time, and can prevent contamination due to long-term exposure in particular.

[0054]

[0055] In one embodiment, the nanocellulose may have an endotoxin content of 5 EU / mL or less.

[0056] Endotoxin is a toxic substance, a potent pyrogen, and can be an indicator of biological safety. The cultured meat scaffold according to the present invention, which is produced by culturing cells on a scaffold and then consumed, requires ensuring biological safety. When endotoxin levels are below the above range, there is no cytotoxicity, resulting in high cell viability during cultured meat production. It is also harmless to the human body, making it suitable as an edible cultured meat scaffold.

[0057]

[0058] In one embodiment, the nanocellulose may have a concentration of 0.1 mg / mL to 10 mg / mL.

[0059] The concentration of nanocellulose can be appropriately selected and used, but preferably, a scaffold with excellent physical properties can be manufactured when the concentration of nanocellulose in the scaffold is within the above range.

[0060]

[0061] According to one embodiment, the polymer hydrogel may include at least one selected from the group consisting of collagen, chitosan, fibrin, agar, starch, alginate, hyaluronic acid, gelatin, dextran, elastin, carrageenan, polyethylene glycol, polyacrylic acid, polyvinyl alcohol, polyvinylpyrrolidone, polylactic acid, and polyglycolic acid.

[0062] Hydrogels contain a large amount of water and have properties between liquids and solids. The polymer chains form a three-dimensional structure and absorb a large amount of water to swell, but due to the crosslink structure, they do not dissolve in water, and can simulate the tissues of actual living organisms. The polymer hydrogel can be used without limitation as long as it is a polymer that can be used in the production of cultured meat. The polymer hydrogel has the advantage of being easy to create a structure because the process of dissolving and shaping the material is simple. Accordingly, it can be used as a three-dimensional cell culture scaffold by improving the problem of two-dimensional cell culture creating an environment different from that of actual tissue in the process of producing cultured meat.

[0063]

[0064] In one embodiment, the polymer hydrogel may be in the form of beads or in bulk form.

[0065] The polymer hydrogel according to the present invention is in the form of beads or bulk, and a polymer hydrogel of an appropriate form can be selected as needed.

[0066]

[0067] According to one embodiment, the weight ratio of the nanocellulose and the polymer hydrogel may be 1:4 to 3:2.

[0068] The weight ratio of the nanocellulose and the polymer hydrogel may preferably be 1:4 to 5:4; 1:4 to 1:1; 1:4 to 3:4; 1:4 to 1:2; 1:2 to 3:2; 1:2 to 5:4; 1:2 to 1:1; 1:2 to 3:4; 3:4 to 3:2; 3:4 to 5:4; 3:4 to 1:1; 1:1 to 3:2; 1:1 to 5:4; or 5:4 to 3:2.

[0069] The pore size and density of the scaffold can be controlled depending on the mixing ratio of the above nanocellulose and the above polymer hydrogel, and when mixed in a weight ratio within the above-mentioned range, a scaffold having excellent physical properties can be manufactured by exhibiting optimal pore size, density, and pore uniformity.

[0070]

[0071] In one embodiment, the scaffold may have a pore diameter of 5 μm to 500 μm, a porosity of 70% to 95%, and a pore uniformity of 10% to 40%.

[0072] The scaffold can form pores by mixing nanocellulose and polymer hydrogel. These pores can regulate the physical properties of the scaffold by participating in the proliferation, differentiation, and adhesion of cultured meat cells. The cultured meat scaffold according to the present invention can increase the uniformity of the pores and maintain the pore shape due to the addition of nanocellulose, thereby increasing metabolism, i.e., nutrient supply and waste excretion.

[0073] The above pore diameter may mean the longest length passing through the center of the pore. Preferably, 5 ㎛ to 400 ㎛; 5 ㎛ to 300 ㎛; 5 ㎛ to 200 ㎛; 5 ㎛ to 100 ㎛; 50 ㎛ to 500 ㎛; 50 ㎛ to 400 ㎛; 50 ㎛ to 300 ㎛; 50 ㎛ to 200 ㎛; 50 ㎛ to 100 ㎛; 100 ㎛ to 500 ㎛; 100 ㎛ to 400 ㎛; 100 ㎛ to 300 ㎛; 100 ㎛ to 200 ㎛; 200 ㎛ to 500 ㎛; 200 ㎛ to 400 ㎛; 200 ㎛ to 300 ㎛; It may be 300 ㎛ to 500 ㎛; 300 ㎛ to 400 ㎛; or 400 ㎛ to 500 ㎛.

[0074] The porosity may preferably be 70% to 90%; 70% to 85%; 70% to 80%; 70% to 75%; 75% to 95%; 75% to 90%; 75% to 85%; 75% to 80%; 80% to 95%; 80% to 90%; 80% to 85%; 85% to 95%; 85% to 90%; or 90% to 95%.

[0075] The above pore uniformity may refer to the difference between the average pore size and each pore size, and the pore size may be 10% to 40% larger or smaller than the average pore size. Preferably, it may be 10% to 30%; 10% to 20%; 20% to 40%; 20% to 30%; or 30% to 40%.

[0076] If the pore diameter is less than the above range, there may be a problem of a decrease in cell attachment rate, and if it exceeds the above range, there may be a problem of not being able to function as a support during cell culture.

[0077] If the porosity is below the above range, there may be a problem of inhibiting cell growth, and if it exceeds the above range, there may be a problem of not being able to function as a support during cell culture.

[0078] If the pore uniformity exceeds the above range, there may be a problem of poor reproducibility during cell culture.

[0079] When a scaffold has an appropriate pore size, porosity, and pore uniformity, the scaffold shape is maintained stably during cell culture, allowing cells to more easily penetrate into the scaffold, and increasing nutrient circulation and oxygen supply, thereby providing an optimal environment for cell growth.

[0080]

[0081] In one embodiment, the scaffold has a density of 1 g / cm 3 10 g / cm 3 , and the strength may be 0.1 MPa to 20 MPa and the endotoxin may be 5 EU / mL or less.

[0082] The above scaffold can provide a scaffold with excellent physical properties due to the mixture of nanocellulose and polymer hydrogel. The physical properties of the scaffold can be expressed as density, strength, toughness, rigidity, cell viability, etc., and the density is preferably 1 g / cm 3 8 g / cm 3 ; 1 g / cm 3 5 g / cm 3 ; 1 g / cm 3 3 g / cm 3 ; 3 g / cm 3 10 g / cm 3 ; 3 g / cm 3 8 g / cm 3 ; 3 g / cm 3 5 g / cm 3 ; 5 g / cm 3 10 g / cm 3 ; or 5 g / cm 3 8 g / cm 3 ; may be, and the strength may preferably be 0.1 MPa to 10 MPa; 0.1 MPa to 5 MPa; 0.1 MPa to 1 MPa; 0.1 MPa to 0.5 MPa; 0.5 MPa to 20 MPa; 0.5 MPa to 10 MPa; 0.5 MPa to 5 MPa; 0.5 MPa to 1 MPa; 1 MPa to 20 MPa; 1 MPa to 10 MPa; 1 MPa to 5 MPa; 5 MPa to 20 MPa; 5 MPa to 10 MPa; or 10 MPa to 20 MPa.

[0083] If the density is below the above range, there may be a problem of poor texture due to the lack of bonding between cells during culture, and if it exceeds the above range, there may be a problem of the cells not maturing sufficiently due to a reduced space for differentiation during culture. The above strength may be a compressive strength, and if the strength is below the above range, there may be a problem of the scaffold collapsing because it cannot function as a support, and if it exceeds the above range, there may be a problem of the cells not growing stably. When the physical properties are within the above range, cells can be cultured stably, easily, and quickly, and the cell survival rate during cell culture may be 80% or more.

[0084] The cultured meat scaffold according to the present invention exhibits optimal density and strength through the addition of nanocellulose, and thus exhibits characteristics that prevent it from easily collapsing even when subjected to physical tensile force. This can help improve the texture of cultured meat by enabling cell alignment and differentiation through the application of tensile force during cultured meat production.

[0085] Cultured meat scaffolds require biological safety, as they are manufactured and consumed as cultured meat. Safety can be demonstrated through the toxic substance endotoxin. If endotoxin levels are below the above range, the scaffold exhibits no cytotoxicity, leading to high cell viability and harmlessness to the human body, making it suitable as an edible cultured meat scaffold.

[0086]

[0087] A method for manufacturing a cultured meat scaffold according to the present invention comprises the steps of: preparing nanocellulose; mixing the nanocellulose with a polymer hydrogel; and freeze-drying.

[0088] The cultured meat scaffold according to the present invention can provide a safe scaffold with excellent physical properties by being manufactured by mixing nanocellulose and polymer hydrogel, thereby manufacturing cultured meat with excellent texture.

[0089]

[0090] According to one embodiment, the step of preparing the nanocellulose may include a step of treating the cellulose at a temperature of 20° C. to 50° C. for 30 minutes to 3 hours; a washing step; and a nanofabricating step.

[0091] The above cellulose may be obtained by preprocessing wood-based or non-wood-based raw materials having a cellulose content of 70% or more using a pulper or refinery, such as by decomposition.

[0092] The above-mentioned treatment step may be to treat pretreated cellulose with TEMPO or an enzyme, or to react untreated cellulose under the above-mentioned conditions. The treatment step may preferably be for 30 minutes to 2 hours, and more preferably for 1 hour to 2 hours.

[0093] If the temperature and time of the above processing step are outside the above range, there may be a problem in producing nanocellulose having mechanical strength and viscosity suitable for the scaffold.

[0094] The above washing step may preferably be performed two or more times.

[0095] The above nano-forming step may be performed by processing at least once with nano-forming equipment such as a high-pressure homogenizer or an ultrasonic emulsifier.

[0096] Cellulose has a large diameter and a low surface area, and poor uniformity between fibers, making it difficult to manufacture a scaffold with a uniform pore size and porosity. However, the nanocellulose obtained by the manufacturing method according to the present invention has a very small diameter and a large surface area, and not only has very high uniformity between fibers, but also has a high porosity, thereby smoothly supplying oxygen and nutrients to cells, thereby making it possible to manufacture a uniform and stable support for cell attachment and growth.

[0097]

[0098] According to one embodiment, the freeze-drying step may be freeze-drying at a temperature of -50°C to -80°C for 48 to 72 hours.

[0099] If freeze-drying is performed below the above temperature and time ranges, the product may not be completely dried, resulting in an unstable or non-uniform shape. If freeze-drying is performed above the above ranges, the product may have different physical properties from the interior due to excessive surface drying. If freeze-drying is performed within the above ranges, not only is moisture completely removed, but the surface and interior can have identical physical properties and structures.

[0100]

[0101] The cultured meat according to the present invention is produced by culturing cells or tissues on a cultured meat scaffold according to the present invention or a cultured meat scaffold produced by a method for producing a cultured meat scaffold according to the present invention.

[0102] Conventional cultured meat has had difficulties in achieving texture and product safety. The cultured meat according to the present invention is cultured by culturing cells or tissues on a cultured meat scaffold comprising nanocellulose and a polymer hydrogel. This cultured meat exhibits high cell or tissue culture activity, enabling easy and rapid culture. Furthermore, the cultured meat exhibits excellent nutrient supply and waste discharge during the culture process, resulting in superior texture and safe cultured meat with improved flavor and texture.

[0103] The above culture may be by seeding or printing cells or tissues.

[0104]

[0105] In one embodiment, the scaffold may be capable of culturing cells or tissues while maintaining a three-dimensional structure.

[0106] Traditionally, cell culture has been performed using two-dimensional (2D) cell culture methods using petri dishes. However, culturing cells using two-dimensional (2D) cell culture methods has problems, such as creating an environment different from that of actual tissue, such as changing cell morphology or allowing cell division to proceed but altering gene expression. Therefore, methods for culturing cells similar to actual tissue using three-dimensional (3D) cell culture methods are being studied, and accordingly, a cell-attachment scaffold with a structure similar to the characteristics of the extracellular matrix is ​​important. Scaffolds manufactured using only a single material have very low cell adhesion, making effective cell growth difficult. The cultured meat according to the present invention utilizes a cultured meat scaffold comprising nanocellulose and a polymer hydrogel to form a network structure by inducing hydrogen bonds between hydroxyl groups on the surface of nanocellulose, thereby maintaining a three-dimensional structure and allowing culture. As a result, effective cell growth can be achieved, resulting in high cell or tissue viability and culture activity, and thus excellent cultured meat can be produced.

[0107]

[0108] In one embodiment, the cell may comprise at least one selected from the group consisting of a stem cell, a bone cell, a muscle cell, an adipocyte, and a stem cell.

[0109] The above cells may be non-human animal derived cells, and are not limited to cells that can be used for producing cultured meat, but are preferably muscle stem cells.

[0110]

[0111] In one embodiment, the cell or tissue survival rate after culture may be 80% or more, and the culture activity may be 20% or more.

[0112] The cultured meat according to the present invention is a method of culturing cells or tissues using a scaffold containing nanocellulose and a polymer hydrogel, and has excellent cell or tissue viability and culture activity during culture, so that cultured meat with excellent texture can be formed easily and quickly.

[0113] The above survival rate may be the ratio of the number of surviving cells after culturing to the number of surviving cells before culturing.

[0114] In the past, the scaffold had poor physical properties, so that it could not maintain its shape or had poor cell adhesion, which prevented smooth metabolism and resulted in low survival rates. However, the cultured meat of the present invention has low cytotoxicity and excellent cell adhesion of the scaffold, and thus can provide cultured meat with excellent physical properties while exhibiting high survival rates and culture activity above the above range.

[0115]

[0116] The cultured meat scaffold according to the present invention comprises nanocellulose and a polymer hydrogel, and a scaffold having excellent physical properties and an optimal pore size and density can be manufactured depending on the ratio of the nanocellulose and the polymer hydrogel according to the present invention, and thus, when manufacturing cultured meat, cell or tissue culture activity and survival rate are excellent, and thus, safe cultured meat with excellent texture and no cytotoxicity can be manufactured easily and quickly.

[0117]

[0118] Hereinafter, the present invention will be described in more detail by way of examples and comparative examples.

[0119] However, the following examples are only intended to illustrate the present invention, and the content of the present invention is not limited to the following examples.

[0120]

[0121] Figures 1a and 1b are images of cultured meat according to examples and comparative examples of the present invention. Referring to Figure 1a, it can be seen that the scaffold cultured meat according to the present invention has a high density and a dense texture, while in the case of the comparative example of Figure 1b, it can be seen that the shape is unstable and the texture is very loose.

[0122] Figures 2a and 2b are images of cultured meat scaffolds according to examples and comparative examples of the present invention. Referring to Figures 2a and 2b, when comparing the example of Figure 2a and the comparative example of Figure 2b, it can be confirmed that the example has a higher density and more uniform pores.

[0123] Figures 3a and 3b are SEM images of cultured meat scaffolds according to examples and comparative examples of the present invention. Figure 3a is a scaffold with a nanocellulose content of 0, and Figure 3b is a scaffold with a weight ratio of nanocellulose and polymer hydrogel of 2.5:7.5. Referring to Figures 3a and 3b, it can be confirmed that the pore size decreases with the addition of nanocellulose, and the pores are uniform.

[0124] Figures 4a and 4b are images confirming cell culture in cultured meat scaffolds according to examples and comparative examples of the present invention. Figure 4a shows the results for the comparative example, and Figure 4b shows the results for the example. Through these images, it can be confirmed that the developed cultured meat scaffold exhibits a high cell growth rate during the same culture period, and that cells grow evenly throughout the scaffold.

[0125] Figure 5 is a graph showing cell viability according to an embodiment of the present invention. To determine cell viability, cell viability was measured using the WST-1 assay. It can be confirmed that the cell viability of cultured meat manufactured using a scaffold manufactured according to the present invention is excellent, and it can be seen that cell viability also increases as the concentration increases.

[0126]

[0127] Although the embodiments have been described above, those skilled in the art will appreciate that various technical modifications and variations can be applied based on the above. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[0128] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

1. Nanocellulose; and Containing a polymer hydrogel; Cultured meat scaffold.

2. In paragraph 1, The above nanocellulose is in the form of powder, suspension or hydrogel. Cultured meat scaffold.

3. In paragraph 1, The above nanocellulose has a diameter of 1 nm to 10 μm. Cultured meat scaffold.

4. In paragraph 1, The above nanocellulose is a hydrogel having a viscosity of 12,000 cP or more, The gelation time is 1 to 30 seconds, Cultured meat scaffold.

5. In paragraph 1, The above nanocellulose has an endotoxin content of 5 EU / mL or less. Cultured meat scaffold.

6. In paragraph 1, The polymer hydrogel comprises at least one selected from the group consisting of collagen, chitosan, fibrin, agar, starch, alginate, hyaluronic acid, gelatin, dextran, elastin, carrageenan, polyethylene glycol, polyacrylic acid, polyvinyl alcohol, polyvinylpyrrolidone, polylactic acid, and polyglycolic acid. Cultured meat scaffold.

7. In paragraph 1, The above polymer hydrogel is in the form of beads or bulk, Cultured meat scaffold.

8. In paragraph 1, The weight ratio of the above nanocellulose and the above polymer hydrogel is 1:4 to 3:

2. Cultured meat scaffold.

9. In paragraph 1, The above scaffold has a pore diameter of 50 μm to 500 μm, The porosity is 70% to 95%, The pore uniformity is 10% to 40%, Cultured meat scaffold.

10. In paragraph 1, The above scaffold has a density of 1 g / cm 3 10 g / cm 3 And, The strength is 0.1 MPa to 20 MPa, Endotoxin is less than 5 EU / mL, Cultured meat scaffold.

11. Step of preparing nanocellulose; A step of mixing the above nanocellulose and polymer hydrogel; and comprising a freeze-drying step; Method for manufacturing cultured meat scaffold.

12. In paragraph 11, The steps for preparing the above nanocellulose are: A step of treating cellulose at a temperature of 20°C to 50°C for 30 minutes to 3 hours; Washing step; and a step of nano-forming; comprising: Method for manufacturing cultured meat scaffold.

13. In paragraph 11, The above freeze-drying step is freeze-drying at a temperature of -50°C to -80°C for 48 to 72 hours. Method for manufacturing cultured meat scaffold.

14. A cultured meat scaffold manufactured by culturing cells or tissues on a cultured meat scaffold manufactured by the method for manufacturing a cultured meat scaffold of paragraph 1 or paragraph 11. Cultured meat.

15. In paragraph 14, The above scaffold maintains a three-dimensional structure and cultures cells or tissues. Cultured meat.

16. In paragraph 14, The above cells include at least one selected from the group consisting of stem cells, bone cells, muscle cells, fat cells, and stem cells. Cultured meat.

17. In paragraph 14, The cell or tissue survival rate after culture is 80% or higher, The culture activity is 20% or more, Cultured meat.

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