Cell culture system, cell culture method, meat production system, and meat production method

Abstract

To provide a low-pressure hypoxic system that can make a low-pressure hypoxia environment capable of controlling a chamber interior to a desired barometric pressure and a desired oxygen concentration according to a storing object, and provide a barometric pressure and oxygen concentration adjustment method, a cell culture system, a cell culture method, a meat production system, a meat production method, a training system, and a training method.SOLUTION: A low-pressure hypoxic system includes: a chamber for accommodating a human, animal and / or material; decompression means for decompressing a barometric pressure in the chamber; and hypoxia concentration control means for maintaining the oxygen concentration in the chamber to the concentration lower than the normal air. The low-pressure hypoxic system is configured such that, in the chamber accommodating the human, animal and / or material, the barometric pressure in the chamber is maintained in a predetermined barometric pressure lower than an ordinary pressure by the decompression means, and the oxygen concentration in the chamber is maintained to the predetermined oxygen concentration of the concentration lower than the oxygen concentration to be decided by the predetermined barometric pressure using the hypoxia concentration control means.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to a low-pressure, low-oxygen system capable of maintaining the air pressure inside a chamber lower than normal pressure and the oxygen concentration lower than that of normal air, a method for adjusting air pressure and oxygen concentration, a cell culture system, a cell culture method, a meat production system, a meat production method, a training system, and a training method. [Background technology]

[0002] So-called high-altitude training has traditionally been practiced as a way to utilize the effects of air pressure and oxygen concentration. As altitude increases, air pressure becomes lower than at sea level (1 atmosphere), and the drop in air pressure reduces the partial pressure of oxygen, resulting in a lower oxygen concentration than normal air (20.9%). In other words, as shown in Figure 1, the oxygen concentration decreases in proportion to the drop in air pressure. For example, it is said that endurance improves when people train in an environment with low air pressure (0.7 atmospheres) and low oxygen concentration (14.6%), equivalent to an altitude of 3,000 m.

[0003] In addition, various factors such as temperature, gas concentration, culture solution composition, and pH affect the growth of animal and plant cells, microorganisms, etc. The oxygen concentration in the culture room is particularly important, and mesenchymal stem cells and other organisms must be cultured under low-oxygen conditions, so various low-oxygen incubators have been developed.

[0004] For example, Patent Document 1 describes a hypoxic incubator that is an airtight culture chamber with a door, and is configured to maintain the O2 concentration at a target constant value by controlling the opening and closing of an N2 concentrator that automatically concentrates N2 in the air and automatic valves V1 and V2 of an N2 storage tank, respectively, by a control unit. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 8-56646 Summary of the Invention [Problem to be solved by the invention]

[0006] However, although the hypoxic incubator in Patent Document 1 adjusts the oxygen concentration by supplying nitrogen gas, it does not take into consideration pressure. Furthermore, because high-altitude training equipment is intended for humans and animals, it does not take into consideration lowering the oxygen concentration even further than before. For this reason, there has been insufficient consideration of the effects of a low-pressure, low-oxygen environment on various things, and of various combinations of air pressure and oxygen concentration.

[0007] The present invention has been made in consideration of the above circumstances, and its purpose is to provide a low-pressure, low-oxygen system, a method for adjusting air pressure and oxygen concentration, a cell culture system, a cell culture method, a meat production system, a meat production method, a training system, and a training method that can easily create a low-pressure, low-oxygen environment in which the air pressure inside the chamber can be controlled to a desired air pressure that is lower than normal pressure and the oxygen concentration can be controlled to a desired oxygen concentration that is lower than normal air, depending on the object to be stored. [Means for solving the problem]

[0008] The low-pressure hypoxic system described in claim 1 comprises a chamber for housing people, animals and / or objects, and a pressure reducing means for lowering the air pressure within the chamber, and is configured so that, when the chamber contains people, animals and / or objects, the pressure reducing means maintains the air pressure within the chamber at a predetermined pressure lower than normal pressure, thereby maintaining the oxygen concentration within the chamber at a predetermined oxygen concentration determined by the predetermined air pressure, which is lower than that of normal air.

[0009] The low-pressure low-oxygen system described in claim 2 comprises a chamber for housing people, animals, and / or objects, a pressure reduction means for lowering the air pressure within the chamber, and a low-oxygen concentration adjustment means for maintaining the oxygen concentration within the chamber at a lower concentration than normal air, and is characterized in that, when the chamber contains people, animals, and / or objects, the pressure reduction means maintains the air pressure within the chamber at a predetermined pressure lower than normal pressure, and the low-oxygen concentration adjustment means maintains the oxygen concentration within the chamber at a predetermined oxygen concentration lower than the oxygen concentration determined by the predetermined air pressure.

[0010] The low-pressure low-oxygen system according to claim 3 is characterized in that the low-oxygen concentration adjusting means includes zeolite.

[0011] The low-pressure low-oxygen system according to claim 4 is characterized in that the pressure inside the chamber is maintained at 0.3 atmospheres or more and less than 1.0 atmospheres.

[0012] The low-pressure, low-oxygen system according to claim 5 is characterized in that the oxygen concentration in the chamber is maintained at 1% or more and less than 21%.

[0013] The low-pressure hypoxic system described in claim 6 is characterized in that a front chamber is provided between the chamber and the outside, and an opening / closing door is provided between the chamber and the front chamber that is configured to close the entrance / exit of the chamber, and the opening / closing door tightly contacts the entrance / exit due to the difference in air pressure between the chamber and the outside of the chamber, thereby closing the entrance / exit.

[0014] The method for adjusting air pressure and oxygen concentration described in claim 7 is characterized in that the air pressure inside a chamber containing people, animals, and / or objects is made lower than atmospheric pressure, and the oxygen concentration is made lower than that of normal air, and the chamber is maintained at a predetermined air pressure lower than atmospheric pressure using a pressure reducing means, thereby maintaining the oxygen concentration inside the chamber at a predetermined oxygen concentration determined by the predetermined air pressure.

[0015] The method for adjusting air pressure and oxygen concentration described in claim 8 is characterized in that the air pressure inside a chamber that houses people, animals, and / or objects is made lower than atmospheric pressure and the oxygen concentration is made lower than normal air, the chamber is maintained at a predetermined air pressure lower than atmospheric pressure using a pressure reducing means, and the oxygen concentration inside the chamber is maintained at a predetermined oxygen concentration lower than the oxygen concentration determined by the predetermined air pressure using a low oxygen concentration adjusting means for maintaining the oxygen concentration inside the chamber lower than normal air.

[0016] The cell culture system described in claim 9 is characterized in that it comprises a low-pressure, low-oxygen system described in any one of claims 1 to 6, and the chamber contains at least one culture chamber that contains a culture container for culturing cells.

[0017] The cell culture system described in claim 10 is characterized in that it comprises a low-pressure, low-oxygen system described in any one of claims 1 to 6, the chamber being housed in a culture room, and the chamber housing a culture container for culturing cells.

[0018] The cell culture system according to claim 11 is characterized in that the culture chamber is an existing cell culture device.

[0019] A cell culture method according to claim 12 is characterized in that the cell culture system according to claim 9 or 11 is used to culture cells in a culture chamber housed in a chamber.

[0020] A cell culture method according to claim 13 is characterized in that the cell culture system according to claim 10 or 11 is used to culture cells in a chamber housed in a culture room.

[0021] A meat production system according to claim 14 comprises the cell culture system according to any one of claims 9 to 11, and is characterized in that it cultures cells that constitute meat.

[0022] The meat production method according to claim 15 is characterized in that meat is produced from cells cultured using the cell culture method according to claim 12 or 13.

[0023] A training system according to a sixteenth aspect of the present invention includes the hypobaric hypoxic system according to any one of the first to sixth aspects of the present invention, and the chamber is a training room.

[0024] A training method according to claim 17 is characterized in that a person and / or an animal exercises in a chamber using the training system according to claim 16. [Effects of the Invention]

[0025] According to the present invention, it is possible to provide a low-pressure, low-oxygen system, a method for adjusting air pressure and oxygen concentration, a cell culture system, a cell culture method, a meat production system, a meat production method, a training system, and a training method, which can easily create a low-pressure, low-oxygen environment inside a chamber that can be controlled to a desired air pressure and oxygen concentration depending on the object to be stored. [Brief explanation of the drawings]

[0026] [Figure 1] FIG. 2 is an explanatory diagram showing the relationship between atmospheric pressure and oxygen concentration. [Figure 2] 1 is an explanatory diagram showing an embodiment of a low-pressure low-oxygen system according to the present invention. [Figure 3] FIG. 10 is an explanatory diagram showing another embodiment of the low pressure low oxygen system according to the present invention. [Figure 4] FIG. 10 is an explanatory diagram showing the results of rearing mice in the same example. [Figure 5] FIG. 10 is an explanatory diagram showing the results of cell culture according to another embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0027] Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. Fig. 1 is a graph showing the relationship between atmospheric pressure and oxygen concentration. Fig. 2 is a schematic diagram showing one embodiment of a hypobaric hypoxic system according to the present invention. Fig. 3 is a schematic diagram showing another embodiment of a hypobaric hypoxic system according to the present invention. Fig. 4 is a graph showing the effect of atmospheric pressure and oxygen concentration on mouse rearing according to the same embodiment. Fig. 5 is a graph showing the effect of atmospheric pressure and oxygen concentration on cell culture according to another embodiment.

[0028] As shown in FIG. 2, the hypobaric hypoxia system 1 according to the present invention comprises a chamber 10 and a pressure reducing means 20. The chamber 10 is used to house a person, an animal, and / or an object. The size of the chamber 10 is not particularly limited, but when housing a person or an animal, it must be large enough to completely fit the person's and / or animal's body, and can range from a capsule-like size that allows the person and / or animal to simply lie down, to a room-like size that allows multiple people and / or animals to sit, exercise, or enter. When housing an object, the size of the chamber 10 can vary depending on the object and amount to be housed, and can be as large as a storage shed, warehouse, greenhouse, incubator, box, or other similar size.

[0029] The pressure reducing means 20 is used to reduce the air pressure inside the chamber 10. The pressure reducing means 20 may be located anywhere inside or outside the chamber 10, but it is preferable that the pressure reducing means 20 is located inside the chamber 10 and pushes the air inside the chamber 10 out of the chamber 10. The pressure reducing means 20 is, for example, a pressure reducing compressor, but may also be other means such as a pump.

[0030] The low-pressure, low-oxygen system 1 may also be equipped with a pressure gauge for monitoring the air pressure inside the chamber 10 and an oxygen concentration meter for monitoring the oxygen concentration. In the present invention, low pressure refers to a value less than 1.0 atmosphere, which is atmospheric pressure (normal pressure), and greater than 0 atmosphere, and a low oxygen concentration refers to a value less than approximately 21%, which is the oxygen concentration in normal air, and greater than 0%.

[0031] The low-pressure, low-oxygen system 1 of the present invention is configured to maintain the air pressure in chamber 10, which contains people, animals, and / or objects, at a predetermined air pressure lower than normal pressure using pressure reduction means 20, thereby maintaining the oxygen concentration in chamber 10 at a predetermined oxygen concentration determined by the predetermined air pressure, which is lower than that of normal air. In other words, by maintaining the air pressure at a predetermined low pressure, the oxygen concentration is determined to be a predetermined oxygen concentration that is proportional to the air pressure. This makes it possible to create a low-pressure, low-oxygen environment that can be controlled to a desired air pressure and desired oxygen concentration. However, the desired oxygen concentration in this case is an oxygen concentration determined by the desired air pressure.

[0032] 3, the low-pressure hypoxic system 2 according to the present invention comprises a chamber 30, a pressure reducing means 40, and a low-oxygen concentration adjusting means 50. The chamber 30 and the pressure reducing means 40 are similar to the chamber 10 and the pressure reducing means 20 of the low-pressure hypoxic system 1.

[0033] The low oxygen concentration adjusting means 50 is used to maintain the oxygen concentration inside the chamber 30 at a concentration lower than the oxygen concentration of normal air (approximately 21%). There are no particular limitations on the configuration of the low oxygen concentration adjusting means 50, but for example, the low oxygen concentration adjusting means 50 is placed at a position where outside air and / or exhaust from inside the chamber 30 passes through, nitrogen is selectively adsorbed onto an adsorbent using a PSA (Pressure Swing Adsorption) method or the like to separate oxygen and nitrogen, air with a high oxygen concentration is discharged, and air with a low oxygen concentration is supplied into the chamber 30.

[0034] For selective adsorption of nitrogen, for example, a vessel filled with zeolite or a hollow fiber membrane made of polyimide or the like is used. Alternatively, oxygen and nitrogen may be separated using an oxygen adsorbent such as a perovskite-type oxide as an adsorbent. By using the low oxygen concentration adjusting means 50, the oxygen concentration in the chamber 30 can be adjusted from a value greater than 0% to less than approximately 21%.

[0035] The low-pressure, low-oxygen system 2 of the present invention is configured so that, in a chamber 30 containing people, animals, and / or objects, the pressure in the chamber 30 is maintained at a predetermined pressure lower than normal pressure by the pressure reduction means 40, and the oxygen concentration in the chamber 30 is maintained at a predetermined oxygen concentration lower than the oxygen concentration determined by the predetermined pressure by the low-oxygen concentration adjustment means 50. In other words, by using the low-oxygen concentration adjustment means 50 in combination with the pressure reduction means 40, the oxygen concentration can be set to a predetermined oxygen concentration lower than a value proportional to the pressure. This makes it possible to create a low-pressure, low-oxygen environment that can be controlled to a desired pressure and a wider range of desired oxygen concentrations other than the oxygen concentration determined by the pressure.

[0036] In the low-pressure, low-oxygen system 2, there are no particular limitations on the configuration of the low-oxygen concentration adjustment means 50, but it is preferable that the low-oxygen concentration adjustment means 50 contain zeolite. A nitrogen adsorber using zeolite, a porous material, can store a large amount of nitrogen even if it is small.

[0037] The air pressure within chambers 10, 30 of low-pressure hypoxic systems 1, 2 must be maintained at a predetermined pressure greater than 0 atmospheres and lower than normal pressure (1 atmosphere) when people, animals, and / or objects are housed therein, but is preferably maintained at a value greater than 0.3 atmospheres and less than 1.0 atmospheres. Furthermore, the oxygen concentration within chambers 10, 30 must be maintained at a predetermined oxygen concentration greater than 0% and lower than the oxygen concentration in normal air (approximately 21%). The oxygen concentration within chamber 30 of low-pressure hypoxic system 2 is preferably maintained at a value greater than 1% and less than 21%.

[0038] The low-pressure hypoxic systems 1, 2 may also be provided with a front chamber 60. Although the front chamber 60 is only shown in FIG. 3, it may also be provided in FIG. 2. The front chamber 60 is provided between the chambers 10, 30 and the outside, and is provided with an opening / closing door 62 configured to close the entrance / exit of the chambers 10, 30 between the chambers 10, 30 and the front chamber 60. A possible configuration is that the opening / closing door 62 tightly contacts the entrance / exit due to the difference in air pressure between the inside and outside of the chambers 10, 30, thereby closing the entrance / exit. This allows the entrance / exit of the chambers 10, 30 to be sealed with a simple structure.

[0039] The hypobaric hypoxic systems 1 and 2 of the present invention can create a hypobaric hypoxic environment. The hypobaric hypoxic environment exerts various effects on the people, animals, and / or objects housed within the chambers 10 and 30, and these effects are thought to vary depending on the people, animals, and objects housed within. For example, in cell culture, some cell types are cultured under hypoxic conditions, but the effects of atmospheric pressure, particularly low pressure, have not been studied to date. Furthermore, even for cells cultured under normal pressure and a normal oxygen concentration, culturing under hypobaric hypoxic conditions may affect cell proliferation and differentiation.

[0040] Therefore, the present invention may be a cell culture system that includes the above-described low-pressure, low-oxygen systems 1, 2 and accommodates cells in chambers 10, 30 for culturing. In this case, chambers 10, 30 accommodate at least one culture chamber 34 that accommodates a culture vessel 32 for culturing cells, and cells are cultured in the culture chamber 34 accommodated in chambers 10, 30. Alternatively, chambers 10, 30 are accommodated in a culture chamber 34, and chambers 10, 30 accommodate a culture vessel 32 for culturing cells, and cells are cultured in chambers 10, 30 accommodated in the culture chamber 34. Then, cells can be cultured in a low-pressure, low-oxygen environment that can be controlled to a desired air pressure and a desired oxygen concentration.

[0041] The culture chamber 34 accommodates culture vessels 32, such as petri dishes or flasks, for culturing cells, and may be of any size as long as it can accommodate the culture vessels 32. The culture chamber 34 may be an existing cell culture device (incubator), but may also be a larger room-like device. The culture chamber 34 may also be formed integrally with the chambers 10 and 30.

[0042] The size of the chambers 10, 30 is also arbitrary. The chambers 10, 30 may be large enough to accommodate one or more culture chambers 34, such as cell culture devices, or may be box-shaped so that the chambers 10, 30 can fit inside the culture chambers 34, such as cell culture devices. Alternatively, the chambers 10, 30 may be large enough to allow a person to enter the chambers 10, 30. If a person can enter the chambers, the person can work inside the chambers 10, 30.

[0043] Furthermore, the cell culture system equipped with the low-pressure, low-oxygen systems 1 and 2 of the present invention may also be used as a meat production system that includes the cell culture system and cultures cells that constitute meat. The meat production system includes the cell culture system, cultures and proliferates cells in a culture vessel 32, differentiates the cells that have proliferated to a confluent state into myotubes, and ultimately produces meat. Note that, in the present invention, "meat" refers not only to cultured meat that is composed of cells, fat, blood vessels, fibers, etc., similar to the meat of animals such as mammals (e.g., pigs and cows), birds (e.g., chickens), and fish, but also to cultured meat that is made up of only muscle cells or primarily muscle cells and formed into a shape similar to animal meat.

[0044] Alternatively, the present invention may be a training system including the above-described hypobaric hypoxic systems 1 and 2. In this case, the chambers 10 and 30 are training rooms in which humans and / or animals exercise. With regard to air pressure and oxygen concentration, the hypobaric hypoxic system 1 of the present invention can maintain a predetermined oxygen concentration determined by a predetermined air pressure, while the hypobaric hypoxic system 2 of the present invention can maintain a predetermined oxygen concentration that is lower than the oxygen concentration determined by the predetermined air pressure. Therefore, the present invention allows training in a hypobaric hypoxic environment in which the desired air pressure and oxygen concentration can be controlled.

[0045] The training rooms, chambers 10 and 30, are large enough to accommodate people and / or animals and allow them to exercise. By placing exercise load devices such as a bicycle ergometer, treadmill, cross trainer, or stepper inside the chambers 10 and 30, the exerciser inside the chambers 10 and 30 can be given an exercise load equivalent to that of cycling, walking, or running.

[0046] By training humans and / or animals in a low-pressure, low-oxygen environment, their oxygen-carrying capacity increases, improving their endurance. Furthermore, by using the low-oxygen-concentration adjusting means 50 to lower the oxygen concentration below that determined by atmospheric pressure, oxygen-carrying capacity is further improved. Of course, in the low-pressure, low-oxygen systems 1 and 2, humans and / or animals can also improve their oxygen-carrying capacity by simply staying in the chambers 10 and 30, although not as much as when they exercise.

[0047] Next, a method for adjusting the air pressure and oxygen concentration in the present invention will be described. The air pressure inside chamber 10, which contains people, animals, and / or objects, is made lower than normal pressure, and the oxygen concentration is made lower than that of normal air, and the inside of chamber 10 is maintained at a predetermined air pressure lower than normal pressure by pressure reducing means 20, whereby the oxygen concentration inside chamber 10 is maintained at a predetermined oxygen concentration determined by the predetermined air pressure lower than normal pressure.

[0048] Another method for adjusting the air pressure and oxygen concentration will now be described. The air pressure inside chamber 30, which contains people, animals, and / or objects, is made lower than atmospheric pressure, and the oxygen concentration is made lower than that of normal air. Chamber 30 is maintained at a predetermined air pressure lower than atmospheric pressure by pressure reduction means 40, and the oxygen concentration inside chamber 30 is maintained at a predetermined oxygen concentration lower than that determined by the predetermined air pressure lower than atmospheric pressure by low oxygen concentration adjustment means 50, which maintains the oxygen concentration inside chamber 30 at a lower concentration than that of normal air.

[0049] In either method, what is done inside the chambers 10, 30 in a low-pressure, low-oxygen environment is optional. For example, as shown in FIG. 3, if the chamber 30 contains a culture chamber 34 that contains a culture vessel 32 for culturing cells, cells may be cultured inside the culture chamber 34. Alternatively, although not shown, if the chambers 10, 30 are contained in the culture chamber 34, cells may be cultured inside the chambers 10, 30. Furthermore, meat may be produced from cells cultured using the cell culture method described above.

[0050] Alternatively, if chambers 10, 30 are training rooms, people and / or animals may exercise in chambers 10, 30, or may stay inside and sleep, read, eat, or otherwise move freely. Furthermore, chambers 10, 30 can be used for a variety of purposes, such as growing plants by adjusting the temperature, humidity, etc., inside a low-pressure, low-oxygen environment, cultivating microorganisms such as koji mold, and storing not only organic matter but also inorganic matter such as metal products.

[0051] The present invention will be described in detail below using examples, but the present invention is not limited to the following examples. [Example]

[0052] [Example 1] In this embodiment, the low-pressure hypoxic system 1 includes a chamber 10 and a pressure reducing means 20, as shown in Figure 2. The chamber 10 is a space large enough to accommodate a person, and capable of maintaining an internal air pressure lower than normal pressure. The pressure reducing means 20 is a pressure reducing compressor disposed within the chamber 10, which pushes the air within the chamber 10 out of the chamber 10 to reduce the air pressure.

[0053] Next, a method for adjusting the air pressure and oxygen concentration of the low-pressure hypoxic system 1 in this embodiment will be described. First, chamber 10 is kept at normal pressure and a normal oxygen concentration, and a target object, such as a person, animal, and / or object, is placed inside chamber 10. Then, pressure reduction means 20 is operated to lower the air pressure inside chamber 10 from normal pressure, and the oxygen concentration becomes lower than that of normal air as the air pressure decreases. By maintaining the air pressure inside chamber 10 at a desired low pressure using pressure reduction means 20, the oxygen concentration inside chamber 10 is also maintained at a predetermined low oxygen concentration determined by the predetermined low pressure, as shown in FIG. 1. In this embodiment, low-pressure hypoxic system 1 was able to adjust the air pressure inside chamber 10 to between 0.3 atmospheres and 1.0 atmospheres, and the oxygen concentration to between 5% and 21%.

[0054] [Example 2] In this embodiment, the low-pressure hypoxic system 2 includes a chamber 30, a pressure reducing means 40, and a low-oxygen concentration adjusting means 50, as shown in Figure 3. The chamber 30 is a space large enough to accommodate a person, and capable of maintaining an internal air pressure lower than normal pressure. The pressure reducing means 40 is a pressure reducing compressor disposed within the chamber 30, which pushes air out of the chamber 30 to lower the air pressure.

[0055] The low-oxygen concentration adjusting means 50 maintains the oxygen concentration in the chamber 30 at a lower concentration than that of normal air, and is a container filled with zeolite that serves as a nitrogen adsorption section. The low-oxygen concentration adjusting means 50 is disposed at a position through which the exhaust gas from the chamber 30 passes by the pressure reducing means 40. By the PSA method, nitrogen gas contained in the exhaust gas is selectively adsorbed onto the zeolite, and air with a high oxygen concentration is discharged outside the chamber 30, and the nitrogen adsorbed onto the zeolite is supplied into the chamber 30.

[0056] 3, the low-pressure hypoxic system 2 includes a front chamber 60 between the chamber 30 and the outside. The front chamber 60 has an opening / closing door 62. The opening / closing door 62 is disposed between the chamber 30 and the front chamber 60.

[0057] Next, a method for adjusting the air pressure and oxygen concentration of the low-pressure, hypoxic system 2 in this embodiment will be described. First, with the chamber 30 at normal pressure and normal oxygen concentration, the door 62 of the front chamber 60 is opened, and a person, animal, and / or object is placed inside the chamber 30. The door 62 of the front chamber 60 is then closed, and the pressure reduction device 40 is operated to lower the air pressure inside the chamber 30 below normal pressure. As the air pressure decreases, the oxygen concentration also becomes lower than normal air. Due to the difference in air pressure between the inside and outside of the chamber 30, the door 62 seals tightly against the entrance to the chamber 30, closing it. The pressure reduction device 40 then maintains the air pressure inside the chamber 30 at a predetermined low pressure lower than normal pressure.

[0058] In addition, by operating the low oxygen concentration adjusting means 50, the oxygen concentration is maintained at a predetermined low oxygen concentration that is even lower than the oxygen concentration determined by the predetermined low pressure shown in Figure 1. In this embodiment, the low pressure low oxygen system 2 was able to adjust the air pressure in the chamber 30 to between 0.3 atmospheres and 1.0 atmospheres, and the oxygen concentration to between 1% and 21%.

[0059] [Example 3] In this example, mice are housed in the chamber 10 of the hypobaric hypoxic system 1 of Example 1. The housed mice are housed in the chamber 10 at room temperature, 0.7 atmospheres, and an oxygen concentration of 14-15% for 8 hours, and then removed from the chamber 10 and housed in an environment at 1.0 atmospheres and an oxygen concentration of 21% for 16 hours. This procedure is repeated for 5 days. After 16 hours of housed mice in a normal environment on the 5th day, the mice's red blood cells (RBC) and hemoglobin (Hb) are measured.

[0060] [Comparative Example] In the comparative example, the mice were reared in the same manner as in Example 3, except that they were reared outside the chamber 10 at 1.0 atmosphere and with an oxygen concentration of 21%, and then the red blood cells (RBC) and hemoglobin (Hb) of the mice were measured.

[0061] As a result, the red blood cell (RBC) value of the mouse was 1048±24×10 in Example 3, as shown in FIG. 4 / μL, and the comparative example is 911±25×10 4 / μL. As shown in FIG. 4(b), the hemoglobin (Hb) value was 16.1±0.4 g / dL in Example 3 and 13.9±0.4 g / dL in the Comparative Example. In Example 3, by rearing the mice under hypobaric hypoxia equivalent to an altitude of 3000 m, the red blood cell (RBC) and hemoglobin (Hb) values ​​of the mice increased by 1.15 times and 1.16 times, respectively, compared to the Comparative Example in which the mice were reared under normal pressure and a normal oxygen concentration, which is the conventional rearing method.

[0062] [Example 4] In this example, cells are cultured in the chamber 30 of the low-pressure, hypoxic system 2 of Example 2. The cell line used is a mouse myoblast cell line, C2C12 cells. Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum is used as the culture medium, and the cells are seeded in a petri dish, which serves as the culture vessel 32. An existing cell culture device (external dimensions: W410 mm x D402 mm x H500 mm, internal capacity: 32 L) is housed within the chamber 30 as the culture chamber 34, and the culture vessel 32 containing the seeded cells is placed within the culture chamber 34. Culture is performed at a temperature of 37°C, with a carbon dioxide concentration of 5%, an oxygen concentration of 10%, and a pressure of 0.7 atmospheres. After 48 hours of culture, trypsinization is performed and the cell count is measured.

[0063] [Comparative Example] In the comparative example, cells were cultured in the same manner as in Example 4, except that the oxygen concentration outside the chamber 30 was 21% and the pressure was 1.0 atmosphere. After culturing for 48 hours, trypsin treatment was carried out and the number of cells was measured.

[0064] As a result, as shown in FIG. 5, the number of cells after 48 hours of culture was 5.65±0.58×10 in Example 4. 5 cell / mL, and the comparison example was 4.05±0.48×10 5 In Example 4, by culturing under low pressure and low oxygen, the cell number increased 1.40 times compared to the comparative example in which the cells were cultured under normal pressure and a normal oxygen concentration, which is the conventional culture method. In a reference example in which cell culture was carried out in the same manner as in Example 4 except that the oxygen concentration was set to 5%, the cells died.

[0065] The low-pressure, low-oxygen systems 1 and 2 of Examples 1 to 4 were able to easily create a low-pressure, low-oxygen environment controlled to the desired air pressure and oxygen concentration. Furthermore, in Example 3, mice were reared in an environment equivalent to an altitude of 3,000 m, and the red blood cell (RBC) and hemoglobin (Hb) values ​​increased. Furthermore, in Example 4, the number of cells increased in the culture of mouse myoblast cell line C2C12 cells, and the low-pressure, low-oxygen system 2 enabled us to investigate conditions suitable for cell proliferation. Therefore, when meat is produced using the cell culture method of the present invention, the speed at which cells proliferate and reach a specific cell number increases, thereby enabling the speed of meat production to be increased.

[0066] The present invention can be used for various purposes other than mouse breeding and cell culture, and it is believed that it will be possible to find objects and conditions that will produce advantageous effects. In addition, in Example 4, by using the low-oxygen concentration adjusting means 50 containing zeolite, it was not necessary to prepare a nitrogen gas cylinder, which has been used until now for cell culture in a low-oxygen environment.

[0067] The present invention allows various embodiments and modifications without departing from the broad spirit and scope of the present invention. Furthermore, the above-described embodiments are intended to explain the present invention and do not limit the scope of the present invention. That is, the scope of the present invention is defined by the claims, not the embodiments. Various modifications made within the scope of the claims and the meaning of the invention equivalent thereto are considered to be within the scope of the present invention. [Industrial Applicability]

[0068] As described above, according to the present invention, it is possible to provide a low-pressure, low-oxygen system, a method for adjusting air pressure and oxygen concentration, a cell culture system, a cell culture method, a meat production system, a meat production method, a training system, and a training method, which can easily create a low-pressure, low-oxygen environment that can be controlled to a desired air pressure and oxygen concentration inside a chamber depending on the object to be stored. [Explanation of symbols]

[0069] 1. Low-pressure hypoxic system 2. Low-pressure hypoxic system 10. Chamber 20. Pressure reducing means 30 Chamber 32...Culture container 34...Cultivation room 40 Pressure reducing means 50. Low oxygen concentration adjustment means 60. Front room 62...Opening and closing door

Claims

1. A chamber for containing cells; a pressure reducing means for reducing the pressure in the chamber; a low oxygen concentration adjusting means for maintaining an oxygen concentration in the chamber lower than that of normal air; A cell culture system characterized in that, while the chamber contains the cells, the pressure reduction means maintains the air pressure in the chamber at a predetermined air pressure lower than normal pressure, and the low oxygen concentration adjustment means maintains the oxygen concentration in the chamber at a predetermined oxygen concentration lower than the oxygen concentration determined by the predetermined air pressure.

2. 2. The cell culture system according to claim 1, wherein the low oxygen concentration adjusting means includes zeolite.

3. 3. The cell culture system according to claim 1, wherein the pressure inside the chamber is maintained at 0.3 atmospheres or more and less than 1.0 atmospheres.

4. 4. The cell culture system according to claim 1, wherein the oxygen concentration in the chamber is maintained at 1% or more and less than 21%.

5. a front chamber is provided between the chamber and the outside; a door between the chamber and the front room configured to close an entrance to the chamber; The cell culture system according to any one of claims 1 to 4, characterized in that the difference in air pressure between the chamber and the outside of the chamber causes the opening and closing door to tightly contact the entrance and close the entrance.

6. A cell culture system described in any one of claims 1 to 5, characterized in that the chamber contains at least one culture chamber that houses a culture container in which the cells are cultured.

7. A cell culture system described in any one of claims 1 to 5, characterized in that the chamber is housed in a culture room and the chamber houses a culture container in which the cells are cultured.

8. A cell culture system described in any one of claims 1 to 5, characterized in that the chamber is made integral with a culture room that houses a culture container in which the cells are cultured.

9. Using the cell culture system according to claims 1 to 8, A cell culture method, comprising culturing the cells in the chamber while the cells are contained therein.

10. A meat production system comprising the cell culture system according to any one of claims 1 to 8, for culturing cells that constitute meat.

11. A method for producing meat from cells cultured using the cell culture method according to claim 9.

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