Cell-cultured food in packaging container, production method for cell-cultured food in packaging container, and preservation method for cell-cultured food in packaging container

Abstract

To provide a cell-cultured food in a packaging container, in which deterioration of the cell-cultured food containing cultured cells can be inhibited.SOLUTION: A cell-cultured food in a packaging container contains cultured cells and is formed by encapsulating the cultured cells in the sealed packaging container, where the inside of the packaging container is in a low oxygen state. Preferably, the packaging container is encapsulated in another sealed packaging container, and a space located inside the other packaging container and outside the packaging container is in a low oxygen state. Furthermore, a deoxygenation agent is preferably encapsulated in the space located inside the other packaging container and outside the packaging container, and a carbon dioxide generation agent is also preferably encapsulated in the space located inside the other packaging container and outside the packaging container.SELECTED DRAWING: Figure 5

Description

[Technical field]

[0001] The present invention relates to a cell-cultured food, and more particularly to a technique for inhibiting deterioration of a cell-cultured food. [Background technology]

[0002] In recent years, technological developments in the production of cell-cultured foods such as cultured meat have progressed. Cell-cultured foods are not only expected to help resolve issues such as food shortages and food waste, but also to reduce livestock production and contribute to preventing global warming. On the other hand, the technology to preserve the quality of cell-cultured foods has not yet been fully developed, and the general understanding is that the quality of cell-cultured foods can be maintained by keeping them in a sterile environment. For example, it is believed that by packaging cell-cultured foods in a sterile environment to prevent quality deterioration caused by microorganisms, the quality of the cell-cultured foods can be maintained without additional sterilization treatments, the addition of food additives, or freezing and storage. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Special Publication No. 2023-519478 [Patent Document 2] Special Publication No. 2022-513441 [Patent Document 3] Special Publication No. 2022-532498 [Non-patent literature]

[0004] [Non-Patent Document 1] Cell Transplantation, Volume 29: 1-7, 2020 [Non-Patent Document 2] Journal of Nutritional Science and Vitaminology 50, 45-49,2004 Summary of the Invention [Problem to be solved by the invention]

[0005] However, a common problem with food preservation is that when food is stored in an atmospheric environment with an oxygen concentration of 21%, the fats and oils, vitamins, colorings, aromatic components, etc. in the food oxidize, impairing the taste of the food. In addition, one of the problems with the preservation of muscle cells in food is that, for example, after only 2 hours of storage in an environment with an oxygen concentration of 1%, the action of superoxide dismutase derived from muscle cells decomposes superoxide anions in muscle cells into oxygen and hydrogen peroxide. It has also been reported that the presence of hydrogen peroxide promotes the activity of proteolytic enzymes, resulting in a decrease in the number of muscle cells (Non-Patent Documents 1 and 2). When proteins contained in food are decomposed in this way, there is a problem of unintended texture and taste.

[0006] Therefore, the inventors conducted intensive research and succeeded in developing a cell-cultured food in a packaging container that can solve the above problems and inhibit deterioration of the cell-cultured food, thereby completing the present invention. Specifically, by enclosing a cell-cultured food containing cultured cells in a sealed packaging container and creating a hypoxic environment inside the packaging container, it was possible to reduce the rate of decrease in the number of viable cells in the cell-cultured food. Furthermore, by sealing the packaging container within another packaging container and creating a hypoxic environment in the space within the other packaging container and outside the packaging container, the rate of decrease in the number of viable cells in the cell-cultured food was also successfully reduced.

[0007] Here, Patent Document 1 describes that the process of packaging and sealing the cultured tissue is carried out under aseptic conditions, so that no additional sterilization procedures or preservative additives are required, and packaged cultured tissue (e.g., meat) with a long shelf life is produced (paragraph 0022). Furthermore, Patent Document 2 describes that, with regard to edible foods that utilize cultured cells, the storage period of the edible foods can be significantly extended because the cells are grown in a completely sterile environment, and that no normal flora (bacteria, yeast, etc.) is present, and that freezing is not necessary due to the aseptic packaging (paragraph 0227). Furthermore, Patent Document 3 states that because the growth process is aseptic, the resulting packaged food is sterile and has a much longer shelf life than conventional fresh products such as fresh meat, and that because the resulting food is sterile, it can be shipped without the need for refrigeration (paragraph 0145).

[0008] However, these patent documents did not state or suggest that maintaining a cell-cultured food containing cultured cells in a hypoxic state would reduce the rate of attrition of cultured cells in the cell-cultured food or inhibit deterioration of the cell-cultured food. The present invention has been made in consideration of the above circumstances, and aims to provide a cell-cultured food in a packaging container that is capable of suppressing deterioration of a cell-cultured food containing cultured cells, a method for producing a cell-cultured food in a packaging container, and a method for preserving a cell-cultured food in a packaging container. [Means for solving the problem]

[0009] In order to achieve the above-mentioned object, the cell culture food in a packaging container of the present invention is a cell culture food in a packaging container containing cultured cells, in which the cultured cells are sealed in a sealed packaging container, and the inside of the packaging container is configured to be in a hypoxic state. It is also preferable that the cell-cultured food in a packaging container of the present invention is configured such that the packaging container is sealed within another packaging container that is sealed, and the space within the other packaging container and outside the packaging container is in a hypoxic state.

[0010] It is also preferable that the cell culture food in a packaging container of the present invention is configured so that an oxygen scavenger is enclosed in the space within the other packaging container and outside the packaging container, and it is more preferable that the oxygen concentration in the space within the other packaging container and outside the packaging container is 0.1% or less.

[0011] It is also preferable that the cell culture food in a packaging container of the present invention is configured so that a carbon dioxide generating agent is sealed in the space within the other packaging container and outside the packaging container, and it is more preferable that the oxygen concentration in the space within the other packaging container and outside the packaging container is 12% or less.

[0012] It is also preferable that the cell culture food in a packaging container of the present invention is configured so that a liquid or gel prepared to have a low oxygen concentration is sealed within the packaging container, or in the space within the other packaging container but outside the packaging container. It is also preferable that the cell culture food in a packaging container of the present invention is configured so that the packaging container has gas permeability and the other packaging container has gas barrier properties.

[0013] It is also preferable that the cell culture food in a packaging container of the present invention is configured so that the packaging container is filled with a liquid or gel. Furthermore, it is also preferable that the cell-cultured food in a packaged container of the present invention be configured by combining the above-mentioned cell-cultured foods in a packaged container in various ways.

[0014] The method for producing a cell-cultured food in a packaging container of the present invention is a method for producing a cell-cultured food in a packaging container containing cultured cells, in which the cultured cells are sealed in a packaging container and the inside of the packaging container is placed in a hypoxic state. In addition, the method for producing a cell-cultured food in a packaging container of the present invention is also preferably a method in which the packaging container is enclosed in another packaging container together with an oxygen scavenger and / or a carbon dioxide generator, and the space within the other packaging container and outside the packaging container is made into a hypoxic state.

[0015] It is also preferable that the method for producing a cell-cultured food in a packaging container of the present invention is a method in which the packaging container is sealed within another packaging container, and the space within the other packaging container and outside the packaging container is made into a low-oxygen state by vacuum packaging. It is also preferable that the manufacturing method of the cell culture food in a packaging container of the present invention is a method in which a liquid or gel prepared to have a low oxygen concentration is enclosed within the packaging container, or within another packaging container in which the packaging container is enclosed, and in the space outside the packaging container.

[0016] The method for preserving a cell-cultured food in a packaging container of the present invention is a method for preserving a cell-cultured food in a packaging container containing cultured cells, in which the cultured cells are sealed in the packaging container and the inside of the packaging container is placed in a hypoxic state. In addition, the preservation method for the cell culture food in a packaging container of the present invention is also preferably a method in which the packaging container is enclosed in another packaging container together with an oxygen scavenger and / or a carbon dioxide generator, and the space within the other packaging container and outside the packaging container is made into a hypoxic state.

[0017] In addition, it is also preferable that the method for preserving a cell-cultured food in a packaging container of the present invention is a method in which the packaging container is sealed within another packaging container, and the space within the other packaging container and outside the packaging container is made into a low-oxygen state by vacuum packaging. It is also preferable that the preservation method for the cell culture food in a packaging container of the present invention is a method in which a liquid or gel prepared to have a low oxygen concentration is enclosed within the packaging container, or within another packaging container in which the packaging container is enclosed, and in the space outside the packaging container. Effect of the Invention

[0018] According to the present invention, it is possible to provide a cell-cultured food in a packaged container that can suppress deterioration of a cell-cultured food containing cultured cells, a method for producing a cell-cultured food in a packaged container, and a method for preserving a cell-cultured food in a packaged container. [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 is a schematic diagram showing the configuration of a cell culture food in a packaging container according to a first embodiment of the present invention. [Diagram 2] FIG. 4 is a schematic diagram showing the configuration of a cell culture food in a packaging container according to a second embodiment of the present invention. [Diagram 3] FIG. 11 is a schematic diagram showing the configuration of an application example of a cell-cultured food in a packaging container according to a second embodiment of the present invention. [Figure 4] FIG. 1 is a diagram showing steps of a test for measuring the number of viable cells in an example. [Diagram 5] FIG. 1 shows the results of a measurement test of viable cell count in an example. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] Hereinafter, embodiments of the cell cultured food in a packaged container, the method for producing the cell cultured food in a packaged container, and the method for preserving the cell cultured food in a packaged container of the present invention will be described in detail. However, the present invention is not limited to the specific contents of the following embodiments and examples described later.

[0021] [First embodiment] The cell culture food in a packaging container of this embodiment is a cell culture food in a packaging container containing cultured cells, characterized in that the cultured cells are sealed in a sealed packaging container and the inside of the packaging container is in a hypoxic state. In the present embodiment, the packaging container is not particularly limited, but a bag-shaped container made of a soft packaging material can be suitably used. For example, a packaging container made of a thermoplastic resin film or sheet can be suitably used.

[0022] Specifically, as shown in FIG. 1, a packaging container 10 having a sealed storage section 12 therein, which is obtained by, for example, heat-welding the peripheral edges 11 of two films, can be used. The cultured cells C are sealed in the storage section 12. In order to prevent the cultured cells C from drying out, it is preferable to use a packaging container 10 in which a liquid or gel L is sealed in the storage section 12. In this embodiment, the shape of the packaging container 10 is not limited to a rectangular shape, and may be any shape such as a polygonal shape, a circular shape, etc. Also, the packaging container 10 does not have to be in a bag shape.

[0023] Cell-cultured foods are foods that contain cultured cells and are also called cellular foods. Cell-cultured foods include so-called cultured meat, as well as various foods that contain cultured cells, such as fish meat, dairy products, sweets, and vegetables made from cultured cells.

[0024] The type of cultured cell C is not particularly limited, but may be one or more animal cells selected from, for example, muscle cells, interstitial cells, adipocytes, chondrocytes, hepatic cells, endothelial cells, renal cells, epithelial cells, nerve cells, lymphatic cells, intestinal cells, cardiac muscle cells, and ciliated epithelial cells. In addition, the cultured cell C can be, for example, one or more plant cells selected from cells of seeds, leaves, stems, roots, immature embryos, mature embryos, callus, microspores, meristems, cotyledons, hypocotyls, epicotyls, mesocotyls, coleoptiles, basal shoots, shoots, or reproductive tissues. Also, cells of other organisms such as insects and microorganisms can be used as the cultured cells C. Furthermore, it is also possible to use a combination of these cells.

[0025] The type of liquid or gel L is not particularly limited, but for example, liquids such as physiological saline or water, gels such as polysaccharides or proteins, etc. can be used.

[0026] A low-oxygen state inside a packaging container means that the oxygen concentration is lower than that in the atmospheric environment, where the oxygen concentration is 21%. Specifically, the oxygen concentration inside the packaging container is preferably 20% or less, more preferably 15% or less, even more preferably 12% or less, even more preferably 10% or less, even more preferably 5% or less, even more preferably 3% or less, even more preferably 2% or less, and particularly preferably 1% or less.

[0027] In this embodiment, the method for creating a low-oxygen state inside the packaging container 10 is not particularly limited, but for example, the low-oxygen state inside the packaging container 10 can be suitably created by forming the packaging container 10 by vacuum packaging. In addition, by sealing a liquid or gel prepared to have a low oxygen concentration inside the packaging container 10, the inside of the packaging container 10 can be suitably made into a low-oxygen state.

[0028] In this embodiment, the material of the packaging container 10 preferably has gas barrier properties. Specifically, the oxygen permeability of the material of the packaging container 10 at 23°C-90% RH is 40cc / m 2 ·day·atm or less is preferable, and 4.4cc / m 2 ·day·atm or less is more preferable, and 0.4cc / m 2 ·day·atm or less is more preferable, and 0.1cc / m 2 It is particularly preferable that the temperature is 100°C or less. As a material for the packaging container 10, for example, a material having an inner layer made of polyethylene with heat sealability, an oxygen absorbing intermediate layer containing an oxygen absorber, and an outer layer with barrier properties can be suitably used.

[0029] The manufacturing method for a cell-cultured food in a packaging container of this embodiment is a manufacturing method for a cell-cultured food in a packaging container containing cultured cells, and is characterized in that the cultured cells are sealed in the packaging container and the inside of the packaging container is placed in a hypoxic state. In addition, the method for preserving a cell-cultured food in a packaging container of this embodiment is a method for preserving a cell-cultured food in a packaging container containing cultured cells, characterized in that the cultured cells are sealed in the packaging container and the inside of the packaging container is kept in a hypoxic state. In the manufacturing method and preservation method of these cell-cultured foods in a packaged container, the cell-cultured food in a packaged container is the same as that described above with reference to FIG.

[0030] According to this embodiment, by using a single packaging container, a cell-cultured food in a packaging container can be obtained in which a hypoxic state is maintained inside the packaging container, and the cell-cultured food in the packaging container can be stored in a state in which a hypoxic state is maintained inside the packaging container. Therefore, with a relatively simple configuration, it is possible to eliminate the problem of the taste of food being impaired when food is stored in an atmospheric environment with an oxygen concentration of 21%, and the problem of the food developing an unintended texture or taste due to oxygen, thereby suppressing the deterioration of cell-cultured foods.

[0031] [Second embodiment] The cell culture food in a packaging container of this embodiment differs from the first embodiment in that the packaging container is sealed inside another sealed packaging container, but the other configurations are the same as those of the first embodiment, except for the points described below. In other words, the cell culture food in a packaging container of this embodiment is a cell culture food in a packaging container containing cultured cells, characterized in that the cultured cells are sealed in a sealed packaging container, the inside of the packaging container is in a hypoxic state, and the packaging container is further sealed within another sealed packaging container, the space within the other packaging container and outside the packaging container is in a hypoxic state.

[0032] Specifically, as shown in Fig. 2, an inner packaging container 10a is enclosed in an outer packaging container 20. The inner packaging container 10a and the outer packaging container 20 can be suitably bag-shaped containers made of soft packaging materials. The inner packaging container 10a can be obtained, for example, by heat-welding the peripheral edges 11a of two films, and has a sealed container section 12a inside.

[0033] Similarly, the outer packaging container 20 can be obtained by heat-welding the peripheral edges 21 of two films, and has a sealed container section 22 inside. In this embodiment, the shapes of the inner packaging container 10a and the outer packaging container 20 are not limited to a rectangular shape, but may be any shape, such as a polygon or a circle.

[0034] The storage section 12a of the inner packaging container 10a is filled with cultured cells C. In order to prevent the cultured cells C from drying and to use the liquid or gel L as a scaffold for culture, it is preferable to fill the storage section 12a with a liquid or gel L together with the cultured cells C.

[0035] Furthermore, in the containerized cell culture food of this embodiment, the space within the outer container 20 and outside the inner container 10a (hereinafter sometimes referred to as inter-container space) is in a hypoxic state. Specifically, the oxygen concentration in the space between the containers is preferably 20% or less, more preferably 15% or less, even more preferably 12% or less, even more preferably 10% or less, even more preferably 5% or less, even more preferably 3% or less, even more preferably 2% or less, and particularly preferably 1% or less.

[0036] In this embodiment, the method for making the inter-container space into a low-oxygen state is not particularly limited, but for example, the inter-container space can be suitably made into a low-oxygen state by forming the outer packaging container 20 by vacuum packaging. Furthermore, by sealing a liquid or gel prepared to have a low oxygen concentration in the space between the containers, the space between the containers can be suitably made into a low oxygen state.

[0037] Furthermore, as an application example of this embodiment, as shown in FIG. 3, it is also preferable to seal an oxygen hypoxic agent 30 in the space between the containers, thereby making the space between the containers into a hypoxic state. As the oxygen reducing agent 30, an oxygen scavenger or a carbon dioxide generating agent can be suitably used. Also, these may be used in combination.

[0038] Specifically, for example, Ageless® (Mitsubishi Gas Chemical Company, Inc.) can be suitably used as the oxygen scavenger. By using such an oxygen scavenger, oxygen in the space between the containers is absorbed by the oxygen scavenger, making it possible to maintain the oxygen concentration in the space between the containers at 0.1% or less, and making it possible to turn most of the remaining gas in the space between the containers into nitrogen, an inert gas.

[0039] As a carbon dioxide generating agent, for example, Anaeropack® (Mitsubishi Gas Chemical Company, Inc.) can be suitably used. By using such a carbon dioxide generating agent, the space between the containers is filled with the carbon dioxide generating agent, and the oxygen concentration in the space between the containers can be maintained at 6 to 12%, and the carbon dioxide concentration can be maintained at 5 to 8%, and most of the remaining gas in the space between the containers can be turned into inert nitrogen gas.

[0040] In this embodiment, the material of the inner packaging container 10a is preferably gas permeable. Specifically, the oxygen permeability of the material of the inner packaging container 10a at 23°C-90%RH is 41cc / m 2 ·day·atm or more is preferable, and 200cc / m 2 ·day·atm or more is preferable, and 1000cc / m 2 ·day·atm or more is more preferable, and 5000cc / m 2 It is particularly preferable that the temperature is 100°C or higher. As the material for the inner packaging container 10a, for example, low-density polyethylene, polymethylpentene, silicone, etc. can be suitably used.

[0041] In this embodiment, the material of the outer packaging container 20 preferably has gas barrier properties. Specifically, the oxygen permeability of the material of the outer packaging container 20 at 23°C-90% RH is 40cc / m 2 ·day·atm or less is preferable, and 4.4cc / m 2 ·day·atm or less is more preferable, and 0.4cc / m2 ·day·atm or less is more preferable, and 0.1cc / m 2 It is particularly preferable that the temperature is 100°C or less. As a material for the outer packaging container 20, for example, a material having an inner layer made of polyethylene with heat sealability, an oxygen absorbing intermediate layer containing an oxygen absorber, and an outer layer with barrier properties can be suitably used.

[0042] The method for producing a cell-cultured food in a packaging container of this embodiment is a method for producing a cell-cultured food in a packaging container containing cultured cells, which is a method in which the cultured cells are sealed in a packaging container and the oxygen concentration in the packaging container is made to be hypoxic, and further includes any one of the following steps:

[0043] That is, it is preferable to enclose a packaging container within another packaging container, and to make the space within the other packaging container and outside the packaging container (inter-container space) into a low-oxygen state by vacuum packaging. It is also preferable to enclose a liquid or gel that has been adjusted to a low oxygen concentration within the packaging container or in the space between the containers. Furthermore, it is also preferable to enclose the packaging container together with an oxygen scavenger and / or a carbon dioxide generator within another packaging container to make the space between the containers hypoxic.

[0044] Furthermore, the method for preserving a cell-cultured food in a packaging container of this embodiment is a method for preserving a cell-cultured food in a packaging container containing cultured cells, which comprises sealing the cultured cells in the packaging container and creating a hypoxic state inside the packaging container, and further comprises any one of the following steps:

[0045] That is, similar to the manufacturing method of the cell-cultured food in a packaging container of this embodiment, it is preferable to enclose the packaging container within another packaging container and make the space within the other packaging container and outside the packaging container (the space between the containers) into a low-oxygen state by vacuum packaging. It is also preferable to enclose a liquid or gel that has been adjusted to a low oxygen concentration within the packaging container or in the space between the containers.

[0046] Furthermore, it is also preferable to enclose the packaging container together with an oxygen scavenger and / or a carbon dioxide generator within another packaging container to make the space between the containers hypoxic. In the method for preserving a cell cultured food in a packaging container according to the present embodiment, the preservation temperature is preferably about 5° C. The temperature suitable for cell preservation is generally about −60 to −80° C. The temperature suitable for cell culture is generally about 37° C. In these methods for producing and storing a cell-cultured food in a packaged container, the cell-cultured food in a packaged container is similar to that described above with reference to FIG. 2 or FIG.

[0047] According to this embodiment, the space inside the other packaging container and outside the packaging container can be maintained in a hypoxic state, and thus it is possible to maintain the inside of the packaging container in a hypoxic state. In other words, by using two packaging containers, it is possible to obtain a cell-cultured food contained in a packaging container in which a hypoxic condition is maintained inside the packaging container, and it is also possible to store the cell-cultured food contained in the packaging container in a condition in which a hypoxic condition is maintained inside the packaging container.

[0048] In this way, this embodiment can use a relatively simple method to solve the problem of food losing its taste when stored in an atmospheric environment with an oxygen concentration of 21%, and the problem of food developing an unintended texture or taste due to oxygen, thereby making it possible to suppress deterioration of cell-cultured foods. EXAMPLES

[0049] Tests conducted to confirm the effects of the packaged cell-cultured food of the present invention, the method for producing the packaged cell-cultured food, and the method for preserving the packaged cell-cultured food of the present invention will be described below. First, two types of packaging containers, an inner packaging container and an outer packaging container, were prepared, and cultured cells and a culture solution were sealed in the inner packaging container. One was made with an oxygen-lowering agent sealed in the space between the containers, which was inside the outer packaging container and outside the inner packaging container, and the other was made without sealing in the oxygen-lowering agent. Then, the number of viable cells in the cultured cells in the inner packaging container after a certain period of time had passed was measured.

[0050] Specifically, a bag made of linear low-density polyethylene (LLDPE) that had been subjected to corona treatment was used as the inner packaging container. The oxygen permeability of the inner packaging container at 23°C-90% RH was 8900cc / m 2 The dimensions are 50 mm x 50 mm. The thickness of the inner packaging film is 100 μm. In addition, a bag made of a film of 12μ PET / 15μ nylon / 70μ CPP was used as the outer packaging container. The 12μ PET layer constitutes the outer surface of the outer packaging container. The oxygen permeability of the outer packaging container at 23℃-90% RH is 0cc / m 2 The dimensions are 110 mm x 160 mm. The thickness of the outer packaging film is 97 μm.

[0051] The cultured cells used were mouse-derived muscle cells (C2C12, KAC Corporation). The culture medium used was DMEM high glucose (Fujifilm Wako Pure Chemical Industries, Ltd., model number 043-30085) containing 10% FBS (Cosmo Bio Co., Ltd., model number S-FBS-NL-025).

[0052] Tests were then conducted using the cell culture food in a packaged container of Comparative Example 1, which did not contain an oxygen hypoxic agent, the cell culture food in a packaged container of Example 1, which contained a carbon dioxide generating agent as an oxygen hypoxic agent, and the cell culture food in a packaged container of Example 2, which contained an oxygen scavenger as an oxygen hypoxic agent, according to the process shown in Figure 4.

[0053] First, a cell suspension containing differentiated cultured cells (mouse-derived muscle cells) and culture solution were injected into the inner packaging container and sealed (step 10). The amount of cultured cells seeded into the inner packaging container was 1.24 million cells / bag. The amount of culture solution injected into the inner packaging container was 4 mL. Then, it was stored at 37° C. for one day (step 11). This allowed the cultured cells to adhere to the inner surface of the inner packaging container.

[0054] Next, the culture medium in the inner packaging container was drained and replaced with 1.9 mL of Dulbecco's phosphate buffered saline (Nacalai Tesque, Inc., model number 14249-95) and 0.1 mL of 2% gelatin solution (Nippi Corporation, model number 892303) (step 12). The inner packaging container was then sealed in the outer packaging container (step 13). At this time, no oxygen hypoxic agent was sealed in Comparative Example 1, while a carbon dioxide generating agent, Anaeropack® (Mitsubishi Gas Chemical Company, Inc.), was sealed in Example 1, and an oxygen scavenger, Ageless® (Mitsubishi Gas Chemical Company, Inc.), was sealed in Example 2.

[0055] In this way, the inside of the outer packaging container was maintained in a hypoxic state (Examples 1 and 2) or not in a hypoxic state (Comparative Example 1), and after storage at 5°C for 6 days (step 14), the inner packaging container was removed from the outer packaging container and the viable cell count of the cultured cells in the inner packaging container was measured (step 15). The viable cell count was measured as follows.

[0056] First, the Dulbecco's phosphate buffered saline was removed from the inner packaging container and the bag was washed once with Dulbecco's phosphate buffered saline (Nacalai Tesque, Inc., model number 14249-95). Next, 0.4 mL of Actase (Nacalai Tesque, Inc., model number 12679-54) was soaked into the entire bag, after which all the liquid was removed and the bag was stored in a CO2 incubator (Sanyo Electric Co., Ltd., model number MCO18AIC) at 37°C for 4 minutes.

[0057] Next, the cells were detached using 1 mL of DMEM high glucose (Fujifilm Wako Pure Chemical Industries, Ltd., model number 043-30085) containing 10% FBS (Cosmo Bio Co., Ltd., model number S-FBS-NL-025), and the detached cells were collected in a 15 mL tube (Corning, model number 430791). Furthermore, 0.1 mL was sampled from the solution collected in the 15 mL tube and applied to a 1.5 mL Eppendorf tube.

[0058] Then, several tens of μL of cell liquid was sampled using a cell counting cartridge (Chemometec, model number 941-0012), and the cartridge was set into a cell counting device (Chemometec, model number NC-200) to measure the cell count and calculate the number of viable cells in the inner packaging container.

[0059] The viable cell count was measured three times for Comparative Example 1, two times for Example 1, and three times for Example 2, and the average was calculated for each. The results are shown in FIG. In addition, the O2 and CO2 concentrations in the space between the containers in Examples 1 and 2 in Figure 5 are based on the product data of the oxygen-lowering agent used and are not actual measurements. Also, the O2 and CO2 concentrations in Comparative Example 1 are in a general atmospheric environment and are not actual measurements.

[0060] As shown in FIG. 5, the viable cell count in Comparative Example 1, in which the space between the containers was not made hypoxic, was 2.1 (10,000 cells / bag). In contrast, the number of viable cells in Example 1, in which the oxygen concentration in the space between the containers was kept at a hypoxic state of 6 to 12%, was 5.2 (10,000 cells / bag), while the number of viable cells in Example 2, in which the oxygen concentration in the space between the containers was kept at a hypoxic state of 0.1% or less, was 210,000 cells / bag.

[0061] Thus, when cultured cells were stored at 5°C for six days using a cell-cultured food in a packaged container, the number of live cells could be doubled by creating a hypoxic state in the space between the containers with an oxygen concentration of 12% or less, compared to when the space between the containers was not created in a hypoxic state. This demonstrates that the rate of decrease in the number of live cells in the cell-cultured food can be reduced, and deterioration of the cell-cultured food can be inhibited, using this cell-cultured food in a packaged container.

[0062] Furthermore, by creating a hypoxic state with an oxygen concentration of 0.1% or less in the space between the containers, the number of live cells could be increased by 10 times compared to when the space between the containers was not created in a hypoxic state. This demonstrates that the rate of decrease in the number of live cells in the cell-cultured food in this packaged container can be further reduced, and deterioration of the cell-cultured food can be significantly suppressed.

[0063] The present invention is not limited to the above-described embodiment and examples, and various modifications can be made within the scope of the present invention. For example, the present invention can be modified appropriately by using a method other than those described above as a means for creating a low-oxygen state in the space between the containers. [Industrial Applicability]

[0064] The present invention can be suitably used in cases such as when preserving cell-cultured foods while suppressing deterioration. [Explanation of symbols]

[0065] 10 Packaging containers 10a Inner packaging container 11,11a Periphery 12,12a Storage section 20 Outer packaging container 21 Periphery 22 Storage unit 30. Oxygen-reducing agents (O2 absorbents, CO2 generators) C Cultured cells L Liquid or gel

Claims

1. A cell culture food in a packaging container containing cultured cells, The cell culture food in a packaged container is characterized in that the cultured cells are sealed in a sealed packaging container, and the inside of the packaging container is in a hypoxic state.

2. 2. The cell-cultured food in a packaging container according to claim 1, characterized in that the packaging container is enclosed within another sealed packaging container, and the space within the other packaging container and outside the packaging container is in a hypoxic state.

3. 3. The cell-cultured food in a packaging container according to claim 2, wherein an oxygen scavenger is enclosed in a space within the another packaging container and outside the packaging container.

4. 4. The cell-cultured food in a packaging container according to claim 3, wherein the oxygen concentration in the space within the another packaging container and outside the packaging container is 0.1% or less.

5. 3. The cell-cultured food in a packaged container according to claim 2, wherein a carbon dioxide generating agent is enclosed in a space within the another package container and outside the package container.

6. 6. The cell-cultured food in a packaging container according to claim 5, wherein the oxygen concentration in the space within the another packaging container and outside the packaging container is 12% or less.

7. 3. The cell-cultured food in a packaging container according to claim 2, characterized in that a liquid or gel adjusted to a low oxygen concentration is sealed inside the packaging container or in a space inside the other packaging container but outside the packaging container.

8. 3. The cell-cultured food in a packaging container according to claim 2, wherein the packaging container has gas permeability and the other packaging container has gas barrier properties.

9. 3. The cell culture food in a packaged container according to claim 1, wherein the packaged container is filled with a liquid or a gel.

10. A method for producing a cell-cultured food in a packaged container, comprising: A method for producing a cell-cultured food in a packaged container, comprising sealing the cultured cells in a packaged container and creating a hypoxic state inside the packaged container.

11. The method for producing a cell-cultured food in a packaging container according to claim 10, characterized in that the packaging container is enclosed in another packaging container together with an oxygen scavenger and / or a carbon dioxide generator, thereby making the space within the other packaging container and outside the packaging container into a hypoxic state.

12. The method for producing a cell-cultured food in a packaged container according to claim 10, characterized in that the packaging container is sealed within another packaging container, and the space within the other packaging container and outside the packaging container is made into a low-oxygen state by vacuum packaging.

13. The method for producing a cell-cultured food in a packaging container according to claim 10, characterized in that a liquid or gel adjusted to a low oxygen concentration is enclosed in the packaging container, or in a space outside the packaging container within another packaging container in which the packaging container is enclosed.

14. A method for preserving a cell-cultured food in a packaged container, which contains cultured cells, comprising: A method for preserving a cell-cultured food in a packaging container, comprising sealing the cultured cells in the packaging container and creating a hypoxic state inside the packaging container.

15. The method for preserving a cell-cultured food in a packaging container according to claim 14, characterized in that the packaging container is enclosed in another packaging container together with an oxygen scavenger and / or a carbon dioxide generator, thereby making the space within the other packaging container and outside the packaging container into a hypoxic state.

16. 15. The method for preserving a cell-cultured food in a packaging container according to claim 14, characterized in that the packaging container is sealed within another packaging container, and the space within the other packaging container and outside the packaging container is made into a low-oxygen state by vacuum packaging.

17. The method for preserving a cell-cultured food in a packaging container according to claim 14, characterized in that a liquid or gel adjusted to a low oxygen concentration is enclosed in the packaging container, or in a space outside the packaging container within another packaging container in which the packaging container is enclosed.

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