Edible support for cell culture of cultured meat and method for manufacturing same

Abstract

The present invention relates to an edible support for the cell culture of cultured meat and a method for manufacturing same and, more specifically, to: an edible support for the cell culture of cultured meat, wherein the edible support can be eaten together with the cultured meat and has a meat texture; a method for manufacturing same; a method for manufacturing cultured meat using same; and a processed food thereof.

Description

Edible support for cell culture of cultured meat and method for producing the same

[0001] The present invention relates to an invention that can provide a support for culturing edible cultured meat cells, a method for producing the same, a method for producing cultured meat using the same, and a processed food product thereof.

[0002] As concerns about environmental pollution caused by the livestock industry and the sanctity of life grow, the number of people seeking cultured meat (or meat substitutes) is increasing worldwide. Cultured meat is a meat substitute that replaces natural meat, and refers to meat that is grown by culturing stem cells in a culture medium to produce lean meat. Many products that utilize textured vegetable protein (TVP) to create existing meat substitutes have been released on the market, but the problem when tasting these products is that they do not reproduce the texture of actual meat.

[0003] Replacing the texture of meat requires a high level of technology, and thus has been considered a difficult problem to date. One way to mimic the texture of meat is to create texture in alternative meats. To achieve this, the American company Beyond Meat has proposed a method of extruding isolated proteins from soybeans. The key technology is to control the pressure, temperature, and speed during extrusion to create dense isolated proteins. Accordingly, significant investment is being made worldwide in the development of extruders, and research is being conducted on technologies that can precisely control high pressure. However, implementing the desired meat texture from plant-based proteins through these methods remains a significant challenge.

[0004] In particular, there have been various attempts to create a texture similar to meat using TVP materials.

[0005] In general, supports made of non-edible raw materials used in the production of cultured meat require a process of separating the cultured cells after cell proliferation and culture. However, there is a problem in that cell destruction occurs during separation, resulting in a decline in the quality of the final cultured meat.

[0006] In addition, in the case of supports manufactured using conventional edible raw materials, there is a limitation in that they are manufactured using hydrogel and 3D printing using gel raw materials, but in terms of texture, they have a jelly-like texture and do not have the texture of meat, and in the case of supports utilizing TVP, there was a problem that the range of applications was very narrow because the strength decreased during the cell culture period and the shape of the support could not be maintained.

[0007] The present invention provides an edible support for cell culture of cultured meat that satisfies the properties required as a support used for producing cultured meat, is edible, has a meat texture, and does not require separate separation of cultured cells from the support, a method for producing the same, and a method for producing cultured meat using the same.

[0008] Accordingly, the technical problems to be solved by the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by a person having ordinary skill in the technical field pertaining to the present invention from the description below.

[0009] The present invention, which aims to solve the above problem, relates to an edible support for cell culture of cultured meat, comprising: an extrudate obtained by extruding textured vegetable protein (TVP) raw material and water; and a surface of the extrudate comprising a collagen-modified layer.

[0010] As a preferred embodiment of the present invention, the TVP raw material may be used that contains 20.50 to 22.50 wt% of carbohydrates, 62.20 to 66.50 wt% of crude protein, 1.50 to 2.00 wt% of crude fat, and the remaining amount of moisture and ash.

[0011] As a preferred embodiment of the present invention, the extrudate may have a length of 3 mm or more and a cross-sectional diameter of 9 mm for the long side and 7 mm for the short side before hydration, and may have a length of 3 mm or more and a cross-sectional diameter of 10 mm for the long side and 8 mm for the short side after hydration. In this case, the hydration is performed for 24 hours under the condition of immersing in water at 20 to 27°C and stirring at 50 rpm using a vertical mixer.

[0012] As a preferred embodiment of the present invention, the edible support has a length of 3 mm and a cross-sectional area of ​​251 mm. 2 At that time, the total average area of ​​pores in the edible support was 10,000 ㎛. 2 It could be strange.

[0013] As a preferred embodiment of the present invention, the total average area of ​​pores in the hydrated edible support measured after hydrating the edible support for 24 hours can increase by 10.0% or less.

[0014] As a preferred embodiment of the present invention, when the compressive strength is measured by compressing the edible support to 75% of its length or height using a CTX measuring device from Brookfield, the compressive strength (TA hardness) may be 100 to 300 g.

[0015] As a preferred embodiment of the present invention, the compressive strength measured after hydrating the edible support for 21 days may have a compressive strength reduction rate of less than 76.0% according to the following mathematical formula 1.

[0016] [Mathematical Formula 1]

[0017] Compressive strength reduction rate (%) = (Compressive strength of edible support before hydration - Compressive strength of edible support after 21 days of hydration) / (Compressive strength of edible support before hydration) × 100%

[0018] In mathematical expression 1, the compressive strength was measured by compressing the edible support to 75% of the length or height using Brookfield's CTX (CTX TEXTURE ANALYZER), and the hydration was performed for 21 days under the condition of immersing the edible support in water at 20 to 27°C and stirring at 50 rpm using a vertical mixer.

[0019] As a preferred embodiment of the present invention, the edible support is capable of satisfies the following equation 1 when measuring the cell attachment rate of muscle stem cells to the edible support after suspending and culturing muscle stem cells under conditions of 37°C and 5% by volume of CO2.

[0020] [Equation 1]

[0021] 500% ≤ (cell attachment rate on day 14 / cell attachment rate on day 7) × 100% ≤ 800%

[0022] As a preferred embodiment of the present invention, the edible support can be configured such that when the cell attachment rate of muscle stem cells to the edible support is measured after suspending and culturing muscle stem cells under conditions of 37°C and 5% by volume of CO2, the cell attachment rate increase rate can satisfy the following equation 2.

[0023] [Equation 2]

[0024] 600% ≤ (cell attachment rate on day 21 / cell attachment rate on day 14) × 100% ≤ 1200%

[0025] As a preferred embodiment of the present invention, the collagen modified layer may be formed by surface modification treatment with a surface modification agent including collagen powder having a molecular weight of 10 to 250 kDa and acetic acid at a concentration of 0.05 to 1.0 volume%.

[0026] As a preferred embodiment of the present invention, the collagen concentration in the surface modifier may be 5 to 100 μg / ml.

[0027] As a preferred embodiment of the present invention, the collagen powder of the surface modifier may include at least one selected from bovine skin-derived collagen, pig skin-derived collagen, chicken skin-derived collagen, and fish-derived collagen.

[0028] Another object of the present invention relates to a method for manufacturing the edible support described above, which can be manufactured by performing a process including: a first step of preparing a tissue vegetable protein (TVP) raw material; a second step of performing an extrusion process with the TVP raw material and water to manufacture an extrudate; and a third step of coating the extrudate with a surface modifier, followed by drying at 30 to 40°C and surface washing with a PBS (phosphate buffer saline) buffer solution to form a collagen-modified layer on the surface of the extrudate.

[0029] As a preferred embodiment of the present invention, the extrudate of the second step may have a length of 3 mm or more and a cross-sectional diameter of 9 mm for the long side and 7 mm for the short side before hydration, and may have a length of 3 mm or more and a cross-sectional diameter of 10 mm for the long side and 8 mm for the short side after hydration. In this case, the hydration is performed for 24 hours under the condition of immersing in water at 20 to 27°C and stirring at 50 rpm using a vertical mixer.

[0030] As a preferred embodiment of the present invention, the surface modifier of step 3 may include collagen powder having a molecular weight of 10 to 250 kDa and acetic acid at a concentration of 0.05 to 1.0 volume%.

[0031] Another object of the present invention is to provide a method for producing cultured meat by culturing cells on the edible support described above.

[0032] In addition, another object of the present invention is to provide cultured meat cultured on the edible support described above; and a cultured meat processed food product that is edible using both the edible support.

[0033] The cultured meat culture support of the present invention is edible and has a texture similar to meat, so that there is no need to separate the cultured cells from the support after cell culture is completed, and since the support of the present invention has a low rate of decrease in compressive strength even after hydration, it not only has excellent shape stability when performing a cell culture process, but also has an excellent cell attachment rate.

[0034] The effects of the present invention are not limited to those mentioned above, and effects not mentioned will be clearly understood by those skilled in the art from the following description.

[0035] Figure 1 is a photograph of an edible support manufactured in Example 1.

[0036] Figure 2, A, is a photograph of an edible support manufactured in Comparative Example 1, and Figure 2, B, is a photograph of a hydrated edible support.

[0037] Figure 3 is a photograph of the edible support manufactured in Comparative Example 2-1.

[0038] Figure 4, A, is a photograph of an edible support manufactured in Comparative Example 2-2, and Figure 4, B, is a photograph of a hydrated edible support.

[0039] Figure 5, A, is a photograph of an edible support manufactured in Comparative Example 3, and Figure 5, B, is a photograph of a hydrated edible support.

[0040] Figure 6, A, is a photograph of an edible support manufactured in Comparative Example 4, and Figure 6, B, is a photograph of a hydrated edible support.

[0041] Figure 7 is data for measuring cell proliferation rates using edible supports of Example 1, Comparative Example 2-1, and Comparative Example 3, conducted in Experimental Example 3.

[0042] In the present invention, “textured vegetable protein (TVP) raw material” means a raw material (or material) of an extrudate used in an extrusion process, and textured vegetable protein (TVP) means an extrudate processed from the raw material.

[0043] The numerical ranges described in the present invention include all values ​​within the range of start and end points, and for example, the range of "20°C to 100°C" means that the values ​​of '20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C, 100°C' as well as any value between integers that fall within the scope of the described range, such as '20°C to 40°C, 50°C to 70°C, 80°C to 100°C'. Additionally, the range of "10 minutes to 100 minutes" is meant to include all integers including, for example, '10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 60 minutes to 70 minutes, 70 minutes to 80 minutes, 80 minutes to 90 minutes, 90 minutes to 100 minutes, as well as any value between integers that fall within the range described in this manner, such as '20 minutes to 40 minutes, 80 minutes to 100 minutes.

[0044] Hereinafter, the present invention will be described in more detail through a method for manufacturing an edible support for cultured meat of the present invention.

[0045] The edible support of the present invention is manufactured by performing a process including: a first step of preparing a textured vegetable protein (TVP) raw material; a second step of performing an extrusion process with the textured vegetable protein raw material together with water to manufacture an extrudate; and a third step of coating the extrudate with a surface modifier, followed by drying and surface washing to form a collagen-modified layer on the surface of the extrudate.

[0046] The TVP raw material of step 1 may include at least one selected from among isolated soy protein powder, concentrated soy protein powder, isolated soy protein powder, textured wheat protein powder, mung bean protein powder, fava bean protein powder, chickpea protein powder, lupin protein powder, lentil protein powder, wheat gluten powder, isolated sunflower seed protein powder, pumpkin protein powder, rice protein powder, hemp seed protein powder, oat protein powder, potato protein powder, and corn protein powder, and preferably may include at least one selected from among isolated soy protein powder, concentrated soy protein powder, isolated soy protein powder, textured wheat protein powder, fava bean protein powder, and chickpea protein powder, and more preferably may include at least one selected from among isolated soy protein powder, concentrated soy protein powder, and isolated soy protein powder.

[0047] And, the TVP raw material may be used that contains 20.50 to 22.50 wt% of carbohydrates, 62.20 to 66.50 wt% of crude protein, 1.50 to 2.00 wt% of crude fat, and the remaining balance of moisture and ash, preferably 21.00 to 22.30 wt% of carbohydrates, 62.50 to 66.00 wt% of crude protein, 1.60 to 1.95 wt% of crude fat, and the remaining balance of moisture and ash, and more preferably 21.00 to 22.00 wt% of carbohydrates, 62.50 to 65.00 wt% of crude protein, 1.60 to 1.90 wt% of crude fat, and the remaining balance of moisture and ash, which is advantageous in terms of securing appropriate strength of the edible support as an extrudate and maintaining strength after hydration.

[0048] Next, in Step 2, the prepared TVP raw material is fed into an extruder and extruded to produce a scaffold-shaped extrudate. At this time, only the TVP raw material is fed into the extruder, and no other materials are used.

[0049] In addition, the extruder may be a general extruder used in the art, and for example, a single-screw extruder, a twin-screw extruder, etc. may be used.

[0050] And, as a preferable example of the above extrusion process, it is performed by feeding TVP raw material into the feeding hopper of a twin-screw extrusion molding machine at a supply amount of 6.0 to 12.0 kg / hr, preferably 7.0 to 10.0 kg / hr. And, the supply amount of water fed into the barrel of the twin-screw extrusion molding machine is performed while feeding 3.50 to 5.00 kg / hr, preferably 3.80 to 4.90 kg / hr, more preferably 4.00 to 4.70 kg / hr, and the water fed is water having a temperature of 20 to 25°C. In addition, it is preferable that the die temperature of the extrusion molding machine be 140 to 170°C, preferably 145 to 160°C, more preferably 145 to 155°C.

[0051] At this time, it is advantageous to perform the extrusion process so that the TVP raw material supply amount, water supply amount, water temperature, and die temperature range satisfy the above ranges, in terms of ensuring that the extrudate or the edible support manufactured using the same has the compressive strength of meat texture and maintains the appropriate compressive strength after hydration. For example, if the water supply amount exceeds 5.00 kg / hr, there is a problem that the porosity of the extrudate increases too much and the compressive strength is greatly reduced, and if the water supply amount is less than 3.50 kg / hr, the compressive strength is high, but there may be a problem that it goes beyond the appropriate compressive range after hydration. In addition, when the die temperature of the extruder increases, the compressive strength of the edible support, which is the extrudate, tends to increase after hydration, but it is advantageous to have the die temperature within the above range in terms of ensuring the appropriate compressive strength after hydration.

[0052] And, the shape of the extrudate can be manufactured into various shapes such as a cylindrical shape, a column shape with a polygonal cross-section, etc. by adjusting the die hole part of the extruder, and the length and / or cross-sectional diameter of the extrudate can also be freely adjusted. The extrudate is in the shape of a scaffold, and in the present invention, the scaffold means an extrudate having a length of 3 mm or more and a cross-sectional diameter of a long side of 9 mm or more and a short side of 7 mm or more before hydration, and an extrudate having a length of 3 mm or more and a cross-sectional diameter of a long side of 10 mm or more and a short side of 8 mm or more after hydration, which is porous and has a network structure inside. At this time, the hydration is performed for 24 hours under the condition of immersing the extrudate in water at 20 to 27°C and stirring it at 50 rpm using a vertical mixer.

[0053] Next, step 3 is a process of surface-modifying the surface of the extrudate manufactured in step 2 with collagen.

[0054] The surface modifier of step 3 includes collagen powder and acetic acid, and can be manufactured by dissolving collagen powder in acetic acid, and the surface modifier can be manufactured by dissolving collagen powder in acetic acid so that the collagen concentration becomes 5 to 100 ㎍ / ml, preferably so that the collagen concentration becomes 5 to 60 ㎍ / ml, and more preferably so that the collagen concentration becomes 7 to 30 ㎍ / ml. At this time, if the collagen concentration in the surface modifier is less than 5 ㎍ / ml, there may be a problem that the amount of collagen fixed to the surface of the surface-treated extrudate is small, and thus the cell attachment rate may be significantly reduced, and even if the collagen concentration exceeds 100 ㎍ / ml, not only will there be no further effect of increasing the cell attachment rate, but rather, there may be a problem that cell attachment is inhibited and cell proliferation rate is low, which is uneconomical.

[0055] And, among the surface modifier components, it is advantageous in terms of cell culture to use collagen powder having a molecular weight of 300 kDa or less, preferably collagen powder having a molecular weight of 10 to 250 kDa.

[0056] In addition, the collagen powder may include at least one selected from collagen derived from bovine skin, collagen derived from pig skin, collagen derived from chicken skin, and collagen derived from fish.

[0057] In addition, the concentration of the acetic acid among the surface modifier components is appropriately 0.05 to 1.0 volume%, preferably 0.05 to 0.5 volume%, and more preferably 0.08 to 0.20 volume%.

[0058] In addition, it is preferable to perform the third-stage coating treatment through a dipping method in which the extrudate is immersed in a surface modifier, or a spraying method in which the surface modifier is sprayed onto the surface of the extrudate.

[0059] And, in the third step, after coating as above, it is dried at 30 to 40℃, preferably 35 to 40℃, to form a coating layer, and then washed with PBS (phosphate buffer saline) buffer solution.

[0060] The edible support for cell culture of the cultured meat of the present invention manufactured by the above method is a porous scaffold.

[0061] The edible support of the present invention was immersed in water at 20 to 27°C and hydrated for 24 hours under the condition of stirring at 50 rpm using a vertical mixer, and had a length of 3 mm and a cross-sectional area of ​​251 mm. 2 At one time, the total average area of ​​pores in the hydrated edible support was 10,000 ㎛. 2 It may be more than 500,000㎛, preferably 2 It may be ideal, more preferably 1,200,000 to 2,300,000 ㎛ 2 It could be.

[0062] And, the total average area of ​​pores in the hydrated edible support increases by 10% or less, preferably by 4.0% to 10.0%, and more preferably by 4.0% to 6.5%.

[0063] Also, 3mm in length and 251mm in cross-sectional area 2 When the hydrated edible support is compressed to 75% of the length or height using a Brookfield CTX measuring device to measure the compressive strength, the compressive strength may be 100 to 300 g.

[0064] In addition, the edible support has a low compressive strength reduction rate after hydration, and the compressive strength measured after hydrating the edible support for 21 days may have a compressive strength reduction rate of less than 76.0% according to the following mathematical formula 1, preferably 70 to 76%, and more preferably 70.0 to 75.5%.

[0065] [Mathematical Formula 1]

[0066] Compressive strength reduction rate (%) = (Compressive strength of edible support before hydration - Compressive strength of edible support after 21 days of hydration) / (Compressive strength of edible support before hydration) × 100%

[0067] In mathematical expression 1, the compressive strength was measured by compressing the edible support to 75% of the length or height using Brookfield's CTX, and the hydration was performed for 21 days under the condition of immersing the edible support in water at 20 to 27°C and stirring it at 50 rpm using a vertical mixer.

[0068] In addition, the edible support of the present invention has an excellent cell attachment rate, and muscle stem cells are suspended and cultured under conditions of 37°C and 5% by volume of CO2, and the edible support (length 3 mm and cross-sectional area 251 mm) 2 ) when measuring the cell adhesion rate, the cell adhesion rate increase rate can satisfy the cell adhesion rate of Equation 1 below.

[0069] [Equation 1]

[0070] 500% ≤ cell adhesion increase rate ≤ 800%, preferably 550% ≤ cell adhesion increase rate ≤ 780%, more preferably 600% ≤ cell adhesion increase rate ≤ 750%

[0071] In Equation 1, the cell adhesion increase rate means (cell adhesion rate on the 14th day / cell adhesion rate on the 7th day) × 100%.

[0072] In addition, the edible support of the present invention is made by suspending and culturing muscle stem cells under conditions of 37°C and 5% by volume of CO2, and forming an edible support (3 mm in length and 251 mm in cross-sectional area). 2 ) when measuring the cell adhesion rate, the cell adhesion rate increase rate can satisfy the cell adhesion rate of Equation 2 below.

[0073] [Equation 2]

[0074] 600% ≤ cell adhesion increase rate ≤ 1200%, preferably 750% ≤ cell adhesion increase rate ≤ 1200%, more preferably 800% ≤ cell adhesion increase rate ≤ 1150%

[0075] In Equation 2, the cell adhesion increase rate means (cell adhesion rate on day 21 / cell adhesion rate on day 14) × 100%.

[0076] Cultured meat can be produced by culturing various cells using the edible support of the present invention manufactured by the method described above. Suitable cells that can be cultured include muscle stem cells, mesenchymal stem cells, adipose stem cells, fibroblasts, embryonic stem cells, induced pluripotent stem cells, vascular stem cells, and / or animal cells.

[0077] In addition, culture conditions, culture medium used for culture, etc. vary depending on the cultured cells, and culture can be performed by selecting culture conditions and culture medium according to the generally known cultured cells.

[0078] The edible support of the present invention has an appropriate strength and can maintain a strength that can have a meat-like texture even after hydration, and can provide various cultured meat processed foods in which the edible support and the cultured meat can be eaten together without separating the cultured meat from the support after producing cultured meat by culturing cells on the edible support.

[0079]

[0080] The present invention is described in detail through preferred embodiments, and the following examples are illustrative of the present invention, but the present application is not limited to the following examples.

[0081] [Example]

[0082] Example 1: Preparation of edible support

[0083] IFF's SPC (ALPHAR) ®12) was prepared using tissue vegetable protein (TVP) as a raw material, and the components of SPC (Soy Protein Concentrate) are shown in Table 1 below.

[0084] Next, the TVP raw material is fed into the feeding hopper of a twin-screw extruder, and then extrusion is performed under the extrusion conditions of a TVP raw material supply amount of 7.50 kg / hr, a water supply amount of 4.62 kg / hr through a barrel, and a die temperature of 158°C to manufacture an extrudate, which is then cut to a length of 3 mm and a cross-sectional area of ​​approximately 198 mm. 2 A TVP, which is a cylindrical extruded product, was manufactured. The number of holes, hole diameter, and hole length (land) of the die were 2 holes and Φ3 mm * 2 mm.

[0085] Next, the TVP, which is the cylindrical extrudate, was immersed in a surface modifier at 37°C for 60 minutes, then taken out and dried at 35-36°C and surface washed with PBS buffer solution to produce an edible support having a collagen-modified layer formed on the surface of the extrudate. At this time, the surface modifier was prepared by dissolving collagen powder (bovine collagen) having a molecular weight of 200-250 Da in a 0.1% by volume acetic acid aqueous solution to a final concentration of 10 ug / ml.

[0086] And, a photo of the manufactured edible support is shown in Fig. 1.

[0087]

[0088] Comparative Example 1

[0089] An edible support is manufactured in the same manner as in Example 1, but without surface modification, and has a length of 3 mm and a cross-sectional area of ​​about 198 mm. 2 A cylindrical extruded product (pre-hydration size) was manufactured as a support. And, its photograph is shown in Fig. 2A.

[0090]

[0091] Comparative Example 2-1

[0092] Using the commercially available IFF product 4380, an edible support having the same shape and size was manufactured by surface modification using the same method as in the example, and this is shown in Fig. 3.

[0093]

[0094] Comparative Example 2-2

[0095] An edible support was manufactured using the same extrudate as in Comparative Example 2-1, but the extrudate without surface modification was manufactured as an edible support in the same shape and size, and a photograph thereof is shown in A of Fig. 4.

[0096]

[0097] Comparative Example 3

[0098] ADM's ISP (product name: ArconS) was prepared as a raw material for tissue vegetable protein (TVP), and the components of ISP (Isolated Soy Protein) are shown in Table 1 below.

[0099] Next, the TVP raw material is fed into the feeding hopper of a twin-screw extruder, and then extrusion is performed under the extrusion conditions of a TVP raw material feed amount of 12.00 kg / hr, a water feed amount of 5.00 kg / hr into the barrel, and a die temperature of 170°C to manufacture an extrudate, which is then cut into pieces having a length of 3 mm and a cross-sectional area of ​​approximately 198 mm. 2 A cylindrical extruded product (pre-weighing size) was manufactured.

[0100] Next, the cylindrical extrudate was immersed in a surface modifier at 37°C for 60 minutes, then taken out and dried at 35-36°C and surface washed with PBS buffer solution to produce an edible support having a modified layer formed on the surface of the extrudate. The surface modifier used was the same as the surface modifier of Example 1.

[0101] A photograph of the manufactured edible support is shown in Fig. 5A.

[0102]

[0103] Comparative Example 4

[0104] An edible support was manufactured using the same method as in Comparative Example 3, but the extrusion molding conditions were performed with a TVP raw material supply amount of 9.00 kg / hr, a water supply amount of 4.54 kg / hr, and a die temperature of 170°C to manufacture an extrudate of the same size, and then surface modification was performed using the same method to manufacture an edible support, the photograph of which is shown in Fig. 6A.

[0105] Classification (g / 100g) TVP raw material analysis Example 1, Comparative Example 1 Comparative Example 3, Comparative Example 4 Carbohydrates 21.55 19.58 Crude Protein 63.2 368.02 Crude Fat 1.72 1.35 Moisture and Ash 13.5 11.05

[0106]

[0107] Experimental Example 1: Compressive Strength Measurement

[0108] The compressive strength before and after hydration of each of the edible supports manufactured in Example 1 and Comparative Examples 1 to 4 was measured by the following method, and the results are shown in Table 2 below.

[0109] Hydration was performed by soaking the edible support in water at 20 to 23°C for 30 minutes, 7 days (1 week), 14 days (2 weeks), and 21 days (3 weeks), and stirring was performed at 50 rpm using the impeller of a mixer.

[0110] And, the microscope photograph of the support of Comparative Example 1 that was hydrated for 24 hours is shown in B of Fig. 2, the microscope photograph of the support of Comparative Example 2-2 is shown in B of Fig. 4, the microscope photograph of the support of Comparative Example 3 is shown in B of Fig. 5, and the microscope photograph of the support of Comparative Example 4 is shown in B of Fig. 6.

[0111] The compressive strength measurement experiment was conducted using Brookefield's CTX equipment (CTX TEXTURE ANALYZER). The ball type probe was moved vertically to measure the height (or length) of the sample. The sample was compressed to 75% of the measured height (or length), and the value measured at this time was measured as the compressive strength.

[0112] And, the compressive strength reduction rate was calculated based on the following mathematical formula 1.

[0113] [Mathematical Formula 1]

[0114] Compressive strength reduction rate (%) = (Compressive strength of edible support before hydration - Compressive strength of edible support after 21 days of hydration) / (Compressive strength of edible support before hydration) × 100%

[0115] Classification Example 1 Comparative Example 1 Comparative Example 2-1 Comparative Example 2-2 Comparative Example 3 Comparative Example 4 TVP Raw Material IFF Company's SPC (ALPHAR ® 12) IFF's 4380ADM's ArconS Surface treatment presence or absence○×○×○○Compressive strength (g) Before hydration809.3809.33867.73867.71283.4Not measurableHydration 30 minutes247.9274.6943.31172.4137.18881.0Hydration 1 week162.9242.4606.5931.9138.4597.3Hydration 2 weeks252.5260.4705.9548.3140.48601.4Hydration 3 weeks202.8180.8996.8579.3Not measurable490.02Compressive strength reduction rate (%), 3 weeks Sign language 74.71% 77.63% 74.22% 85.80% Incalculable Incalculable

[0116] Looking at the compressive strength measurement results in Table 2 above, the compressive strength reduction rate measured after 3 weeks of hydration was less than 75%, showing excellent compressive strength retention. In contrast, Comparative Example 1, which was not surface treated, showed a tendency for the compressive strength reduction rate to be higher compared to Example 1. This tendency could also be confirmed through the compressive strength reduction rates of Comparative Examples 2-1 and 2-2.

[0117] In addition, the appropriate compressive strength for having a meaty texture is 100 to 300 g, preferably 200 to 300 g, after hydration. However, Comparative Example 2-1 had a problem in that the compressive strength measured after 3 weeks of hydration was too high at over 900 g.

[0118] In addition, Comparative Example 3 was completely decomposed in the third week of the hydration experiment, and it is believed that it was completely decomposed due to the impact from the impeller of the mixer.

[0119] In addition, in the case of Comparative Example 4, there was a problem in which the compressive strength before hydration exceeded the measurement limit of 5000 g, which is judged to be a result of inappropriate extrusion process conditions.

[0120]

[0121] Experimental Example 2: Measurement of the total area of ​​pores in an edible support

[0122] The total area of ​​pores in the edible supports manufactured in the above examples and comparative examples was measured before and after hydration by the following method, and the results are shown in Table 3 below.

[0123] The pore area was calculated by manually designating the outline of all pores on the plane of the edible support, photographing three supports once with a Nikon SMZ745T using a low-magnification microscope and the compatible program HKBasic, and designating the area of ​​all observable pore regions in the photograph, and calculating them in the program. However, when selecting pores, they were visually confirmed, and those that were not pores were excluded and measured.

[0124] Example 1 Comparative Example 2-1 Comparative Example 3 Comparative Example 4 Total area of ​​pores in the support before hydration (㎛) 2 )2,084,450Not measurable269,156113,22124 Total pore area within the support after hydration for 4 hours (㎛) 2 )2,182,879362,779219,725 Total pore area increase rate 4.72% 34.78% 84.07%

[0125] As shown in Table 3 above, the edible support of the present invention showed no significant change in the total pore area within the support before and after hydration, showing an area change rate of less than 5%. In contrast, Comparative Examples 3 and 4 showed excessively high area change rates of 34.78% and 84.07%, respectively.

[0126] In addition, Comparative Example 2-1 had a surface formed with micropores, so when photographed with a low-magnification microscope, the pores on the surface were not discernible, and the photograph was taken as a flat plane, making it impossible to measure the entire pore area.

[0127] Through this, it was confirmed that the edible support of the present invention has very high morphological stability.

[0128]

[0129] Experimental Example 3: Measurement of cell attachment rate to edible support

[0130] After the edible supports of Example 1, Comparative Example 2-1, and Comparative Example 3 were externally coated using collagen, the cell attachment rate for each of them was measured, and the results are shown in Table 4 below.

[0131] Cell culture was performed using the 3D cell culture method, which suspends cells in a medium and cultures them. The medium used was DMEM (Dulbeco's Modified Eagle's Medium) containing FBS (Fetal bovine serum). The cells used were mouse myoblasts, C2C12. The cell culture temperature was 37℃ and was performed for 1 week (7 days), 2 weeks (14 days), and 3 weeks (21 days).

[0132] And, the cell attachment rate to the support was measured by detaching the cells from the support and counting them, and the measurement data is shown in Fig. 7, and the unit of the cell attachment rate in Table 4 is %.

[0133] Classification Example 1 Comparative Example 2-1 Comparative Example 3 Week 1 Cell Attachment Rate (%) 40.6% 53.0% 2.7% Number of Cells (Week 1) 20,300 26,500 1,366 Number of Cells (Week 2) 128,333 79,800 41,566 Number of Cells (Week 3) 1,442,333 862,333 634,000 Cell Proliferation Rate (Week 2 Cell Proliferation Rate Compared to Week 1) 6.3 times 3 times 30 times Cell Proliferation Rate (Week 3 Cell Proliferation Rate Compared to Week 2) 11.2 times 10.8 times 15.3 times

[0134] Looking at the cell attachment rate measurement results in Table 4 above, Comparative Example 2-1 showed the highest cell attachment rate, but the lowest cell proliferation rate, and Comparative Example 3 showed the highest cell proliferation rate, although the cell attachment rate was low. Example 1 showed a lower cell attachment rate than Comparative Example 2-1, but a high cell proliferation rate, and although the cell proliferation rate was lower than Comparative Example 3, when the total number of cells was counted, Example 1 showed the most proliferation. This is because the pore size of the support is a factor that ensures the flow of medium into and out of the support and supports the movement of cell waste products. However, in the case of Comparative Examples 2-1 and 3, the flow of medium and the movement of waste products generated during cell growth were insufficient due to the micropores, which caused the differences in cell attachment rate and cell attachment increase rate as described above.

[0135]

[0136] Examples 2 to 5 and Comparative Examples 5 to 8

[0137] Using the TVP raw material used in Example 1, a cylindrical extrudate, TVP, was manufactured in the same manner, and then treated with a surface modifier to form a collagen-modified layer. An edible support having the same size and shape as Example 1 was manufactured, but the extrusion process conditions were changed as shown in Table 5 below, and Examples 2 to 5 and Comparative Examples 5 to 8 were performed.

[0138] Then, the manufactured edible support (extruded product) was immersed in water at 20 to 23°C and stirred at 50 rpm using a vertical mixer for 24 hours, and the compressive strength was measured, and the results are shown in Table 5 below.

[0139] Classification Screw speed (rpm) Number of holes * Hole diameter (mm) * Hole length (mm) Die temperature (℃) Water supply (kg / hr) TVP raw material input (kg / hr) After hydration for 24 hours, compressive strength (g) Compressive strength (g) Week 1 Week 2 Week 3 Example 2500 2 holes * 3 * 2 150 4.37 8 27 7.36 323.12 39 4.84 15 9.66 Example 3500 2 holes * 3 * 2 150 3.98 8 26 7.74 4 29.18 39 5.4 13 0.84 Example 4500 2 holes * 3 * 2 160 4.37 8 8 30.10 8 7 4.48 7 27.12 28 1.08 Example 55002 hole*3*2 1704.909 280.724 24.22 378.1198.32 Comparative example 55002 hole*3*2 1505.16 8920.02 1037.98 896.64 385.30 Comparative example 65002 hole*3*2 1705.839 150.20 229.18 188.62 76.33 Comparative example 75002 hole*3*2 1706.5 210 252.32 178.76 254.32 81.04 Comparative example 85002 hole*2*6 1705.05 12 145.90 176.28 129.94 50.00

[0140] Looking at Table 5 above, it was confirmed that in the case of Examples 2 to 5, the compressive strength after hydration for 3 weeks satisfied the range of 100 to 300 g, which has a meat texture. In contrast, Comparative Example 5, in which the water supply to the barrel of the twin-screw extruder exceeded 5.00 kg / hr, had a problem in which the compressive strength after hydration for 3 weeks exceeded 300 g, and Comparative Examples 6 and 7, in which the TVP raw material input was slightly increased, the die temperature of the extruder was 170°C, and the water supply exceeded 5.00 kg / hr, had a problem in which the compressive strength was significantly reduced even though the water supply was increased compared to Comparative Example 5.

[0141] In addition, in the case of comparative example 8, where the TVP raw material input amount was high, there was also a problem that the compressive strength was significantly low after hydration.

[0142] Through the above examples and experimental examples, it was confirmed that the present invention can provide an edible support for cell culture of cultured meat that satisfies the properties required as a support used for producing cultured meat, is edible, has a meat texture, and does not require separate separation of cultured cells from the support.

Claims

1. An extrudate obtained by extruding textured vegetable protein (TVP) raw material and water, and the surface of the extrudate includes a collagen modified layer. An edible support for cell culture of cultured meat, wherein the extrudate has a length of at least 3 mm and a cross-sectional diameter of at least 9 mm on the long side and at least 7 mm on the short side before hydration, and after hydration for 24 hours has a length of at least 3 mm and a cross-sectional diameter of at least 10 mm on the long side and at least 8 mm on the short side.

2. An edible support for cell culture of cultured meat, wherein the TVP raw material comprises 20.50 to 22.50 wt% of carbohydrates, 62.20 to 66.50 wt% of crude protein, 1.50 to 2.00 wt% of crude fat, and the remaining amount of moisture and ash in the first paragraph.

3. An edible support for cell culture of cultured meat, wherein, after hydrating the edible support for 24 hours in the first paragraph, the total average area of ​​pores in the hydrated edible support increases by 10% or less.

4. In the third paragraph, the hydrated edible support has a length of 3 mm and a cross-sectional area of ​​251 mm. 2 When the total average area of ​​pores in the hydrated edible support is 10,000 ㎛ 2 Edible support for cell culture of cultured meat.

5. An edible support for cell culture of cultured meat, wherein, in the third paragraph, when the compressive strength is measured by compressing the hydrated edible support to 75% of the length or height using a CTX measuring device of Brookfield, the compressive strength (TA hardness) is 100 to 300 g.

6. In the first paragraph, the compressive strength measured after hydrating the edible support for 21 days is an edible support for cell culture of cultured meat, wherein the compressive strength reduction rate according to the following mathematical formula 1 is less than 76.0%; [Mathematical Formula 1] Compressive strength reduction rate (%) = (Compressive strength of edible support before hydration - Compressive strength of edible support after 21 days of hydration) / (Compressive strength of edible support before hydration) × 100% In mathematical expression 1, the compressive strength was measured by compressing the edible support to 75% of the length or height using Brookfield's CTX, and the hydration was performed for 21 days under the condition of immersing the edible support in water at 20 to 27°C and stirring it at 50 rpm using a vertical mixer.

7. In the first paragraph, the edible support is an edible support for cell culture of cultured meat, wherein when the cell attachment rate of muscle stem cells to the edible support is measured after suspending and culturing muscle stem cells under conditions of 37°C and 5% by volume of CO2, the cell attachment rate increase rate satisfies the following equation 1; [Equation 1] 500% ≤ (cell attachment rate on day 14 / cell attachment rate on day 7) × 100% ≤ 800% 8. In the first paragraph, the collagen modified layer is formed by surface modification treatment with a surface modification agent containing collagen powder having a molecular weight of 10 to 250 kDa and acetic acid at a concentration of 0.05 to 1.0 volume%. An edible support for cell culture of cultured meat, wherein the collagen concentration in the surface modifier is 5 to 100 ㎍ / ml.

9. An edible support for cell culture of cultured meat, wherein the collagen powder in paragraph 8 comprises at least one selected from collagen derived from bovine skin, collagen derived from pig skin, collagen derived from chicken skin, and collagen derived from fish.

10. Step 1: Preparing textured vegetable protein raw materials; Step 2: Extruding the above-mentioned vegetable protein raw material with water to produce an extrudate; and A process including the third step of coating the extrudate with a surface modifier, drying it at 30 to 40°C, and washing the surface with a PBS (phosphate buffer saline) buffer solution to form a collagen-modified layer on the surface of the extrudate is performed. A method for producing an edible support for cell culture of cultured meat, wherein the surface modifier comprises collagen powder having a molecular weight of 10 to 250 kDa and acetic acid at a concentration of 0.05 to 1.0 volume%.

11. In the 10th paragraph, the extruded product of the second step has a length of 3 mm or more and a cross-sectional diameter of 9 mm or more on the long side and 7 mm or more on the short side before hydration, A method for producing an edible support for cell culture of cultured meat, which has a length of at least 3 mm and a cross-sectional diameter of at least 10 mm on the long side and at least 8 mm on the short side after hydration for 24 hours.

12. A method for producing cultured meat by culturing cells on an edible support selected from any one of claims 1 to 9.

13. A cultured meat product in which cells are cultured on an edible support selected from any one of claims 1 to 9; and a cultured meat processed food in which all of the edible support are edible.

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