JP6727042B2 - Container with gas supply function, cell culture container, transport container, storage container, cell culture method, content transport method and content storage method - Google Patents

Description

The present invention provides a container having a function of supplying gas into a container, the container having a gas supply function; a cell culture container comprising the container having a gas supply function, a carrying container and a storage container; and a container having a gas supply function. The present invention relates to a cell culture method, a content transport method and a content storage method used.

In order to culture cells efficiently, gas such as oxygen and carbon dioxide may be supplied to the cell culture container (Patent Document 1).
In addition, in order to prevent deterioration of the state of cells when carrying the cultured cells, gas such as oxygen, carbon dioxide and nitrogen may be supplied to the cell carrying container (Patent Document 2).

However, in the conventional cell culture container and cell transport container, it is necessary to supply gas from the outside of the container, so it is necessary to separately prepare a high-pressure gas cylinder or the like. Therefore, there is a problem that the entire apparatus including the container and the high pressure gas cylinder becomes complicated and large. Further, there are many restrictions on the use of high-pressure gas, and there is a problem that the high-pressure gas cylinder cannot be handled easily.

As a transport container and a storage container that do not require a high-pressure gas cylinder or the like, a container in which an atmosphere modifier is stored has been proposed (Patent Document 3).

Japanese Unexamined Patent Application Publication No. 2016-077164 JP, 2005-124556, A JP, 2014-083002, A

However, since the atmosphere modifier uses a chemical reaction, heat and by-products are easily generated. Heat and by-products may contaminate contents such as cells or deteriorate the condition. Further, when a gas atmosphere composed of a plurality of gases is required, the composition of the atmosphere adjusting agent becomes complicated and it is not practical.

INDUSTRIAL APPLICABILITY The present invention has a simple structure, is compact, can be easily handled, and is unlikely to contaminate the contents or deteriorate the condition thereof; a cell culture container comprising the gas supply function container, and a transport container. And a container for storage; and a method for culturing cells, a method for transporting contents, and a method for storing contents using a container with a gas supply function.

The present invention has the following aspects.
<1> A container with a gas supply function, comprising a container body and a gas supplier housed in the container body, wherein the gas supplier has the following gas-containing material.
Gas-containing material: In a substrate comprising either one or both of a liquid composition containing a gelling agent and a liquid medium, and a gel composition containing a network structure formed from the gelling agent and a liquid medium. Including gas in a bubble state.
<2> The container body has a gas permeable partitioning portion that divides the inner space of the container body into a gas supply chamber in which the gas supply body is stored and a content storage chamber in which the content is stored. A container with a gas supply function according to <1> above.
<3> The container with a gas supply function according to <2>, wherein the partition section has a gas permeable film facing the content storage chamber side.
<4> The container with a gas supply function according to <3>, wherein the partition part has a gas-permeable support layer that supports the gas-permeable membrane.
<5> In the above <1> to <4>, the gas contains at least one gas selected from the group consisting of hydrogen, oxygen, nitrogen, argon, helium, carbon dioxide, methane, ethane, propane and butane. Either container with gas supply function.
<6> The content of gas in the gas-containing material is 6 to 80% by volume in terms of volume/mass% (v/w%) of the gas-containing material, according to <1> to <5> above. Either container with gas supply function.
<7> The container with a gas supply function according to any one of <1> to <6>, wherein the gas-containing gas contained in the gas-containing material has a bubble diameter equal to or smaller than a microbubble size.
<8> The <1> to <wherein the gelling agent is at least one selected from the group consisting of gelatin, agar, carrageenan, pectin, glucomannan, pullulan, sodium alginate, and carboxyvinyl polymer or a derivative thereof. A container with a gas supply function according to any of 7>.
<9> A cell culture vessel comprising the vessel with a gas supply function according to any one of <1> to <8>.
<10> A container for transportation, comprising the container with a gas supply function according to any one of the items <1> to <8>.
<11> A storage container comprising the container with a gas supply function according to any one of the above <1> to <8>.
<12> A cell culture method in which cells are cultured using the container with a gas supply function according to any one of <1> to <8>.
<13> A content transportation method for transporting the content using the container with a gas supply function according to any one of the above <1> to <8>.
<14> A method for storing contents, wherein the contents are stored using the container with a gas supply function according to any one of <1> to <8>.

The container with a gas supply function of the present invention has a simple structure, is compact, can be easily handled, and is unlikely to contaminate the contents or deteriorate the condition.
The cell culture container, the transport container and the storage container of the present invention have a function of supplying gas into the container, have a simple structure, are compact, can be easily handled, and contaminate the contents. Or it is hard to make the condition worse.
According to the cell culturing method of the present invention, cells can be cultivated while easily supplying the required gas without preparing a complicated and large-sized device, and the cells are unlikely to be contaminated or deteriorated.
According to the content transporting method of the present invention, the content can be transported while easily supplying the required gas without preparing a complicated and large-sized device, and the content is unlikely to be contaminated or the state thereof is not deteriorated.
According to the content storage method of the present invention, the content can be stored while easily supplying the required gas without preparing a complicated and large-sized device, and the content is unlikely to be contaminated or deteriorated.

It is sectional drawing which shows an example of the container with a gas supply function of this invention. It is sectional drawing which shows a mode that the container main body of the container with a gas supply function of FIG. 1 was decomposed|disassembled. It is sectional drawing which shows the other example of the container with a gas supply function of this invention. It is sectional drawing which shows the other example of the container with a gas supply function of this invention. It is sectional drawing which shows the other example of the container with a gas supply function of this invention. It is sectional drawing which shows a mode that the inside of the container main body was suction-deaerated in the container with a gas supply function of FIG. 3 is a photograph of the appearance of the liquid composition (X1) of Example 1 and the gas-containing materials of Examples 1, 3 to 5, and 7. 3 is a photograph of a cross-sectional microscope observation of the gas-containing material of Example 1. 5 is a graph showing a time change of hydrogen gas concentration in a gas phase by a gas release test from a gas-containing material using a container with a gas supply function in Example 2. 5 is a photograph of a cross-sectional microscope observation of the gas-containing material of Example 3. 8 is a photograph of a cross-sectional microscope observation of the gas-containing material of Example 4. 8 is a photograph of a cross-sectional microscope observation of the gas-containing material of Example 5. 9 is a photograph of a cross-sectional microscope observation of the gas-containing material of Example 6. 9 is a photograph of a cross-sectional microscope observation of the gas-containing material of Example 7. It is a graph which shows the time change of the content of the gas in the gas containing material in a 25 degreeC sealed container in Examples 1, 3-7.

The following definitions of terms apply throughout the specification and claims.
The “gel” refers to one in which a liquid medium is retained in a network structure formed by physical aggregation of a gelling agent by hydrogen bonding or the like or cross-linking of the gelling agent by covalent bonding or the like.
“Physical gel” refers to a liquid medium held in a network structure formed by physical aggregation of a gelling agent by hydrogen bonding or the like.
“Chemical gel” refers to a liquid medium held in a network structure formed by cross-linking a gelling agent by covalent bonding or the like.
"Gelling" means that a liquid composition containing a gelling agent and a liquid medium is changed into a gel composition containing a network structure formed from the gelling agent and a liquid medium.
"Gel melting" means that a gel composition containing a network structure formed from a gelling agent and a liquid medium is changed to a liquid composition containing a gelling agent and a liquid medium.
The “gelling agent” refers to a compound capable of forming a network structure capable of holding a liquid medium by physical aggregation by hydrogen bonding or the like or crosslinking by covalent bonding or the like.
The "bubbles having a size of micro bubbles or less" means bubbles having a diameter of micron order or less. The bubbles may include bubbles having a diameter of nano order.
The “saturated solubility of gas with respect to a substrate” refers to the saturated solubility of gas with respect to the liquid composition under atmospheric pressure when the substrate is entirely a liquid composition. The “gas dissolution” that defines the saturation solubility is a state in which Henry's law is established and the gas is dissolved in a molecular state according to the pressure.

The saturated solubility of a gas in a substrate can be measured by gas chromatography (GC). The temperature at which the saturated solubility is measured needs to be a temperature at which the base material is all a liquid composition, and in the case of a liquid composition, it may be a temperature higher than the gelation temperature, and it reversibly changes to a liquid composition. In the case of a gel composition that can be used, the temperature may be higher than the gel melting temperature.
The content of gas in the gas-containing material (volume/mass% (v/w%) conversion of the gas-containing material) is measured by weighing a sample collected from the gas-containing material under atmospheric pressure and 10° C. The gas contained in the sample is heated to 70° C. to be released into the gas phase, the gas in the gas phase is quantified by gas chromatography (GC), and the amount of gas released from a predetermined mass (100 g) of the gas-containing material is determined. It is obtained by calculating the ratio of volume (cm ³ ). However, carbon dioxide and hydrocarbons are determined by the method described in Examples below.
The gelling temperature is determined as follows. A 50 mL glass screw bottle was charged with a liquid composition having a volume of about half the volume of the screw bottle and the container was sealed and immersed in a constant temperature water bath, heated to 75° C., and then gradually cooled down in the constant temperature water bath, and screw bottle The temperature at which the liquid composition does not flow even when tilted at 45° and 90° is the gelling temperature.

"-" showing a numerical range means including the numerical value described before and after that as a lower limit and an upper limit.
The dimensional ratios in FIGS. 1 to 5 include those different from actual ones for convenience of description. 2 to 5, the same components as those of FIG. 1 are designated by the same reference numerals and the description thereof will be omitted.

<<Vessel with gas supply function>>
FIG. 1 is a sectional view showing an example of a container with a gas supply function of the present invention.
The container 1 with a gas supply function is a container having a gas-permeable partition 12 that divides an internal space into a gas supply chamber 102 in which a gas supply body 50 is stored and a content storage chamber 104 in which a content 200 is stored. The main body 10 and the gas supply body 50 housed in the gas supply chamber 102 of the container body 10 are provided.

<Container body>
FIG. 2 is a cross-sectional view showing a state where the container body 10 is disassembled.
The container body 10 includes a lid body 20, an upper vessel 30 to which the lid body 20 is attached by screwing, and a lower vessel 40 to which the upper vessel 30 is attached by screwing.

The lid 20 has a tubular portion 22 and a canopy portion 24 that seals the upper end of the tubular portion 22. On the inner peripheral wall of the lid 20, a female screw portion 28 that is screwed with the upper male screw portion 36 of the upper container 30 is provided.

The upper container 30 has a tubular portion 32; and a gas permeable partition portion 12 that divides the internal space of the tubular portion 32 into upper and lower parts. An upper male screw portion 36 that is screwed into the female screw portion 28 of the lid 20 is provided on the upper portion of the outer peripheral wall of the tubular portion 22. A lower female threaded portion 38 that is screwed into the upper male threaded portion 46 of the lower container 40 is provided on the lower portion of the inner peripheral wall of the tubular portion 22.

The lower container 40 has a tubular portion 42; and a bottom portion 44 that vertically divides the internal space of the tubular portion 42. An upper male screw portion 46 to be screwed with the lower female screw portion 38 of the upper container 30 is provided on the upper portion of the outer peripheral wall of the tubular portion 42. At the lower part of the inner peripheral wall of the tubular portion 42, a lower female threaded portion 48 that is screwed with an upper male threaded portion of another container when mounting another container is provided.

The partition part 12 has a gas permeable film 14 facing the content storage chamber 104 side; and a gas permeable support layer 16 that supports the gas permeable film 14. The support layer 16 is formed integrally with the tubular portion 32 of the upper container 30, and the support layer 16 has a plurality of through holes 18 formed therein.

The gas permeable film 14 is a layer made of a gas permeable material. The gas permeable film may be one that allows gas released from the gas supply body 50 to permeate from the gas supply chamber 102 to the content storage chamber 104 and does not permeate the content 200 in the content storage chamber 104 to the gas supply chamber 102. Good. Examples of the material of the gas permeable membrane include polydimethylsiloxane, biaxially stretched polystyrene, ethylene-vinyl acetate copolymer, polystyrene and polyethylene. As the material of the gas permeable membrane, polydimethylsiloxane is preferable from the viewpoint of excellent cell adhesion.

Examples of the material of the container body 10 (excluding the gas permeable film 14) include resin, glass, metal (aluminum, iron, alloys containing these, etc.) and the like.
When it is necessary to visually check the gas supply body 50 and the content 200, the material of the container body 10 is preferably resin or glass, and more preferably resin because the container body 10 can be easily manufactured. Examples of the resin include polyethylene terephthalate, polyvinyl chloride, polystyrene, polypropylene and the like.
On the other hand, when container strength is required or when gas permeation loss from the container body 10 is desired to be reduced, stainless steel, aluminum and alloys thereof are preferable as the material of the container body 10, and aluminum is preferable because it is lightweight and easy to process. Duralumin and the like are more preferable.

<Gas supplier>
The gas supplier 50 has a gas-containing material described below. The gas supply body 50 further has a material container (a petri dish, a bottle, a gas permeable film, or a bag made of a metal sheet or a resin sheet processed with holes such as gas vents) that stores the gas-containing material. You may have. In addition, the gas supply body 50 may further include a carrier carrying a gas-containing material.

(Gas-containing material)
The gas-containing material contains a gas in a bubble state in a base material (base material) made of either one or both of a liquid composition described later and a gel composition described later.

The gas-containing material includes a gas dissolved in the base material and a gas in a bubble state contained in the base material. Since the gas-containing material includes gas in a bubble state, in the substrate, the saturated solubility of the gas in the liquid composition when the substrate is entirely a liquid composition (hereinafter, gas for the substrate is (It is also referred to as the saturated solubility of .). That is, the total amount of the gas dissolved in the base material and the gas in the bubble state exceeds the saturated solubility of the gas in the base material. When the amount of gas in the gas-containing material exceeds the saturated solubility of gas in the base material, the gas-containing material can release the amount of gas exceeding the saturated solubility of gas in the base material to the outside.

The content of gas in the gas-containing material can be appropriately selected according to the type of gas, the use of the gas-containing material, etc., within the range that the substrate can hold.
The content of gas in the gas-containing material should be in a wide range of 6 to 80% by volume in terms of volume/mass% (v/w%) of the gas-containing material according to the method for producing the gas-containing material described later. Is possible. When the gas content is at least the lower limit value of the above range, for example, the saturated solubility of hydrogen gas in water is 1.6 ppm (1.7% by volume), so that the saturated solubility of gas in the base material is far higher. Since the amount of gas that exceeds is retained, a large amount of gas can be released. It is physically difficult for the gas content to exceed the upper limit of the above range, and there is concern that the strength of the obtained gas-containing material may be reduced, which is not preferable.

The bubble diameter of the gas in the bubble state is preferably a microbubble size or less, more preferably 200 μm or less, from the viewpoint of uniform dispersion of the gas and gas retention stability in the gas-containing material. When the bubble diameter of the gas in the bubble state is 200 μm or less, the gas in the bubble state can be surely included in the base material, and the gas retention stability in the base material is improved. When the bubble diameter exceeds 200 μm, the buoyancy of the gas itself becomes large during the production of the gas-containing material, the loss of gas outside the system increases, and it becomes difficult to produce the gas-containing material having a large gas content. The gas in the bubble state in the present invention may include bubbles having a nano bubble size (1 to 100 nm) smaller than the micro bubble size. Nanobubble-sized bubbles have improved storage stability in a liquid composition.

The gas content in the gas-containing material and the bubble diameter of the gas in the bubble state can be adjusted by the components of the composition described below, the method of dispersing the gas in the liquid composition in the method of producing the gas-containing material described below, and the like.

(gas)
The gas contained in the gas-containing material and released from the gas-containing material is not particularly limited as long as it is a gas that can exhibit a useful function in the fields of cell culture, medicine, food, cosmetics and the like.

The gas capable of exhibiting a useful function includes at least one gas selected from the group consisting of hydrogen, oxygen, nitrogen, argon, helium, carbon dioxide, methane, ethane, propane and butane. The gas may be a mixed gas. The ratio of each gas in the mixed gas is arbitrary. Air is included in the mixed gas.

As the gas, at least one gas selected from the group consisting of hydrogen, oxygen, nitrogen, argon, helium and carbon dioxide is more preferable, and a gas containing hydrogen is particularly preferable.
Hydrogen is used in the fields of food, cosmetics, medical treatment, etc., which utilize the potential of reducing and antioxidizing properties. In addition, hydrogen can be expected to exhibit cell proliferation ability in cell culture and various cell functions.
Oxygen is used in fields such as cell culture, medical care, and health care equipment.
Since nitrogen, argon, and helium are inert gases in nature, they can be used in the fields of cell culture, food, cosmetics, etc., where antioxidant is required.
Carbon dioxide is used in fields such as cell culture, food, and cosmetics.

Although it is difficult to dissolve hydrogen in a high concentration in the base material, hydrogen can be contained in the base material in a high concentration as a gas in a bubble state by the method for producing a gas-containing material described below.

(Base material)
The base material (base material) of the gas-containing material is composed of either one or both of a liquid composition described below and a gel composition described below.
It is preferable that the substrate contains a gel composition because it can contain a large amount of gas in a bubble state and can release the gas for a long time because the gas in a bubble state can be retained for a long time. More preferably,

(Composition)
The liquid composition includes a gelling agent and a liquid medium.
The gel composition is a gel composition of a liquid composition, and includes a network structure formed from a gelling agent and a liquid medium.
The liquid composition and the gel composition (hereinafter collectively referred to simply as “composition”) may contain a filler and an additive, if necessary, within a range that does not impair the effects of the present invention. Good.

The composition can be reversibly changed from a liquid composition to a gel composition and from a gel composition to a liquid composition (X), and from a liquid composition to a gel composition. , And a composition (Y) which cannot be changed from a gel composition to a liquid composition.
Hereinafter, each of the composition (X) and the composition (Y) will be described.

(Composition (X))
Examples of the liquid composition (X1) in the composition (X) include a composition capable of forming a network structure capable of holding a liquid medium by physical aggregation of the gelling agent by cooling.
Examples of the gel composition (X2) in the composition (X) include a so-called physical gel in which a liquid medium is held in a network structure formed by physical aggregation of a gelling agent. In the gel composition (X2) which is a physical gel, since the network structure is formed by the physical aggregation of the gelling agent, the network structure is easily dissolved by heating. Therefore, the gel composition (X2) can be returned to the original liquid composition (X1) by heating.
Thus, the composition (X) reversibly changes from the liquid composition (X1) to the gel composition (X2) by cooling and from the gel composition (X2) to the liquid composition (X1) by heating. It can change.

The liquid composition (X1) has a gelling temperature which changes from the liquid composition (X1) to the gel composition (X2) by cooling.
The gel composition (X2) has a gel melting temperature that changes from the gel composition (X2) to the liquid composition (X1) by heating.
The gelling temperature of the liquid composition (X1) is preferably 0.5 to 65°C, more preferably 10 to 60°C.
When the gelling temperature is within the above range, the composition (X) can be easily maintained in the gel composition (X2) state in the normal operating temperature range of the container 1 with a gas supply function. Therefore, the base material composed of the composition (X) can contain a large amount of gas in a bubble state. Further, the gas can be released for a long time because the base material can hold the gas in a bubble state for a long time.

Selection of each component contained in the composition (X) may be appropriately performed depending on the application of the container 1 with a gas supply function and the operating temperature range (the required gelling temperature. When used in the fields of medicine, food, cosmetics, etc., in addition to the gas retention performance, storage stability and gelation temperature of the composition (X) of the base material comprising the composition (X), the composition Each component contained in the composition (X) may be appropriately selected in consideration of the safety of (X) against the living body, adverse effects on cells during cell culture, and the like.

Examples of the gelling agent in the composition (X) include proteins or polysaccharides derived from natural products such as gelatin, agar, carrageenan, pectin, glucomannan, pullulan and sodium alginate; polymers having hydrogen-bonding groups; aggregation. And the like having a hydrophobic group having a hydrophilic property. The gelling agents may be used alone or in combination of two or more.
The gelling agent is preferably at least one selected from the group consisting of gelatin, agar, carrageenan, pectin, glucomannan, pullulan and sodium alginate.

The liquid medium is appropriately selected depending on the use of the container 1 having a gas supply function and the type of each component (particularly gelling agent) of the composition (X). For example, when the container 1 with a gas supply function is used in the fields of cell culture, medical treatment, food, cosmetics, etc., water, ethanol, etc. are used as the liquid medium. In applications where human safety is not required, any organic solvent or the like may be used.

(Composition (Y))
Examples of the liquid composition (Y1) in the composition (Y) include a composition capable of forming a network structure capable of holding a liquid medium by crosslinking a gelling agent.
Examples of the gel composition (Y2) in the composition (Y) include a so-called chemical gel in which a liquid medium is retained in a network structure formed by crosslinking a gelling agent. In the gel composition (Y2) which is a chemical gel, since the network structure is formed by the cross-linking of the gelling agent, it is difficult to eliminate the network structure. Therefore, the gel composition (Y2) does not change to the original liquid composition (Y1) due to heating or the like.
Thus, the composition (Y) can be changed from the liquid composition (Y1) to the gel composition (Y2) by the chemical reaction of the gelling agent, but the gel composition (Y2) is changed to the liquid composition ( You cannot change to Y1).

Gelation of the liquid composition (Y1) may be changed so that a network structure is formed by crosslinking and a chemical gel is formed. As a method of gelling the liquid composition (Y1), a method suitable for the liquid composition (Y1) to be used may be selected. Examples of the gelation method include a method of adding a crosslinking agent, neutralization, heating and the like.

Since the composition (Y) cannot be changed from the gel composition (Y2) to the liquid composition (Y1), it is usually used as a composition composed only of the gel composition (Y2). When the composition (Y) consists only of the gel composition (Y2), the base material composed of the composition (Y) can contain a large amount of gas in a bubble state. Further, the gas can be released for a long time because the base material can hold the gas in a bubble state for a long time.

Selection of each component contained in the composition (Y) may be appropriately performed depending on the use of the container 1 with a gas supply function. When the container 1 with a gas supply function is used in fields such as cell culture, medical care, food, cosmetics, etc., in addition to the gas retention performance and storage stability of the base material comprising the composition (Y), the composition Each component contained in the composition (Y) may be appropriately selected in consideration of the safety of (Y) against the living body, adverse effects on cells during cell culture, and the like.

Examples of the gelling agent in the composition (Y) include a polymer having a crosslinkable functional group, a polyfunctional monomer, a monofunctional monomer, a silicone polymer, a carboxyvinyl polymer or a derivative thereof. The gelling agents may be used alone or in combination of two or more.
As the gelling agent, a carboxyvinyl polymer or its derivative is preferable because it is easy to control gelation by a simple operation such as neutralization and concentration adjustment.

The liquid medium is appropriately selected depending on the use of the container 1 having a gas supply function and the type of each component (particularly gelling agent) of the composition (Y). For example, when the container 1 with a gas supply function is used in the fields of cell culture, medical treatment, food, cosmetics, etc., water, ethanol, etc. are used as the liquid medium. In applications where human safety is not required, any organic solvent or the like may be used.

(Additive)
As an additive that can be added to the composition, when the container 1 with a gas supply function is used in the fields of cell culture, medical care, food, cosmetics, etc., for the purpose of expressing a synergistic effect or imparting a new effect, Any known additive can be used as long as it is a component that can be used in combination with each component contained in the composition.

Examples of the additives include food additives, cosmetic additives, antioxidants, medium additives, feed additives and the like. Specifically, the following may be mentioned.
Bactericides: sodium hypochlorite, sodium sulfite, high-grade coconut powder, etc.
Emulsifiers: glycerin fatty acid ester, sucrose fatty acid ester, steariloyl calcium lactate, sorbitan fatty acid ester, propylene glycol fatty acid ester and the like.
Thickening stabilizer: sodium alginate, propylene glycol alginate, propoxymethylcellulose sodium, propoxymethylcellulose calcium, sodium starch glycolate, sodium polyacrylate, methylcellulose, hydroxymethylcellulose, natural polysaccharides and the like.
Water retention emulsion stabilizer: sodium chondroitin sulfate, etc.
Binders and quality modifiers: potassium polyphosphate, sodium polyphosphate, potassium metaphosphate, sodium metaphosphate, etc.
Anti-sticking agent: D-mannitol and the like.
Preservatives: benzoic acid, benzoate, sorbic acid, sorbate, paraoxybenzoic acid esters, sodium dehydroxyacetate, propionic acid, propionate, white protein, polylysine, pectin degradation products and the like.
Antioxidants: erythorbic acid, erythorbate, isopropyl citrate, dibutylhydroxytoluene, dl-α tocopherol, nordihydroguaiaretic acid, butylhydroxyanisole, propyl gallate and the like.
Reinforcing agents: vitamins, etc.
Others: Amino acid derivatives, nucleic acids, lipids, antioxidants, anti-glycation agents, fats and oils, surfactants and the like.
The additives may be used alone or in combination of two or more, depending on the desired effect.

(Method of manufacturing gas-containing material)
A method for producing a gas-containing material will be described by taking a gas-containing material in which a gas in a bubble state is contained in a base material composed of the composition (X) as an example.
The gas-containing material can be produced by, for example, a method including the following steps (1) to (3).
Step (1): While maintaining the liquid composition (X1) at a temperature higher than the gelation temperature, by supplying a gas to the liquid composition (X1), the liquid composition (X1 containing a gas in a bubble state ) Is obtained.
Step (2): A step of filling a liquid container (X1) containing a gas in a bubble state in a filling container and sealing the container.
Step (3): A group consisting of the gel composition (X2) by cooling the liquid composition (X1) containing a gas in a bubble state to a gelation temperature or lower in the filling container to cause gelation. A step of obtaining a gas-containing material in which a gas in a bubble state is included in the material.

The method including the steps (1) to (3) is a suitable method for producing a gas-containing material. According to the method including the steps (1) to (3), a gas-containing material having a large gas content can be stably produced, as described below.
Usually, the gas is more likely to move in the liquid composition (X1) as the viscosity of the liquid composition (X1) is lower, and is more easily dispersed as fine bubbles. However, the fact that the gas easily moves in the liquid composition (X1) means that it is difficult for the gas to stay in the liquid composition (X1) and the gas is likely to be vaporized into the gas phase. On the contrary, when the liquid composition (X1) has a high viscosity, the gas is difficult to disperse, which is not preferable for forming fine bubbles. Of course, in the gel composition (X2) obtained by gelling the liquid composition (X1), it becomes substantially impossible to disperse the gas.
According to the method including the steps (1) to (3), the gas is uniformly dispersed in the liquid composition (X1) as fine bubbles to a high concentration, and then the liquid composition (X1) is rapidly cooled. By gelling, a gas in a bubble state can be included at a high concentration in the base material composed of the gel composition (X2).

(Preparation of liquid composition (X1))
The liquid composition (X1) is prepared, for example, by charging a liquid medium at room temperature into a dissolution tank, charging the gelling agent under stirring, and then raising the temperature to a temperature at which the gelling agent can be dissolved and dissolving it in the liquid medium. Can be prepared.

Examples of the apparatus used for preparing the liquid composition (X1) include a tank or a kettle equipped with a stirrer. The material of the device can be selected within the range that does not impair the effects of the present invention, in consideration of corrosion resistance against gelling agent, liquid medium, gas, etc.; heat resistance at use temperature; elution into liquid composition (X1) and the like. .. Examples of the material of the device include stainless steel material, glass lining, fluororesin lining, plastic and the like.

(Process (1))
By supplying gas to the liquid composition (X1) while maintaining the liquid composition (X1) at a temperature higher than the gelation temperature, the liquid composition (X1) containing gas in a bubble state is obtained.

The viscosity of the liquid composition (X1) during gas supply is preferably 5 to 1000 mPa·s, more preferably 10 to 800 mPa·s. When the viscosity of the liquid composition (X1) is less than the lower limit value of the above range, the gas is likely to be dispersed as fine bubbles in the liquid composition (X1), but the gas easily floats and is hard to stay in the liquid. When the viscosity of the liquid composition (X1) exceeds the upper limit of the above range, it is difficult to disperse the gas in the liquid composition (X1) as fine bubbles, and it is difficult to uniformly disperse the gas.
On the other hand, the temperature of the liquid composition (X1) at the time of supplying the gas may be any temperature as long as it can achieve the viscosity range of the liquid composition (X1) described above, and may be appropriately selected.

The amount of gas supplied is such that the total amount of the gas dissolved in the liquid composition (X1) and the gas in the bubble state exceeds the saturated solubility of the gas in the liquid composition (X1). The content of gas in the gas-containing material obtained in Step 1 is preferably 6 to 80 volume% in terms of volume/mass% (v/w%) of the gas-containing material.

The charging amount of the gelling agent and the liquid medium, and the supply amount of the gas may be appropriately set according to the desired gas content in the gas-containing material. Also, the temperature and viscosity of the liquid composition (X1) may be appropriately set according to the type of the gelling agent, the liquid medium and the gas, and the content of the gas in the desired gas-containing material.

As a device used in the step (1), a device and equipment used in a known gas-liquid dispersion operation can be used as long as the device can uniformly disperse gas in the liquid composition (X1) as fine bubbles. The material of the device can be selected within the range that does not impair the effects of the present invention, in consideration of corrosion resistance against gelling agent, liquid medium, gas, etc.; heat resistance at use temperature; elution into liquid composition (X1) and the like. ..

In the step (1), gas is supplied to the liquid composition (X1) while the liquid composition (X1) is being stirred; or the liquid composition (X1) is not stirred. It is preferable to shake after feeding the gas to ().
Examples of the stirring method include a method using a stirrer and a method using a line mixer.
Examples of the shaking method include a method using a shaker.

According to the method using the stirrer, gas is introduced from the lower part of the stirring blade while stirring the liquid composition (X1), and the gas can be finely dispersed in the liquid composition (X1) by stirring.
Examples of the apparatus equipped with a stirrer include vessels such as turbine blades suitable for gas dispersion in a liquid, a kettle equipped with stirring blades such as full-zone blades, and tanks.

According to the method using a line mixer, the liquid composition (X1) and gas are introduced into the line mixer, and the gas can be finely dispersed in the liquid composition (X1) by line mixing.
Examples of the line mixer include known turbo mixers, static mixers, ejectors, and other devices suitable for fine mixing of gas and liquid.

In the method using a shaker, it is possible to use a sealed shaker in which the contents are less likely to leak during shaking.
The liquid composition (X1) is charged into the shaker to about 1/2 volume of the shaker volume. In order to replace the gas phase part of the shaker with gas, 1 to 5 times the gas phase volume of the shaker was introduced into the liquid composition (X1) after bubbling or into the gas phase, Cover and seal. The closed shaker is shaken until the gas in the gas phase is finely dispersed in the liquid composition (X1).

(Process (2))
A liquid composition (X1) containing gas in a bubble state is filled in a filling container and sealed.
In order to suppress the volatilization loss of gas in a bubble state into the gas phase, it is preferable to perform filling and sealing as quickly as possible.

The temperature of the liquid composition (X1) at the time of filling is preferably 5 to 20° C. higher than the gelation temperature. When the temperature of the liquid composition (X1) at the time of filling is at least the lower limit value of the above range, it is easy to increase the content of gas in a bubble state contained in the liquid composition (X1). When the temperature of the liquid composition (X1) at the time of filling is equal to or lower than the upper limit value of the above range, the gas in the bubble state contained in the liquid composition (X1) is less likely to be vaporized into the gas phase.

The viscosity of the liquid composition (X1) at the time of filling is preferably 100 to 10,000 mPa·s. When the viscosity of the liquid composition (X1) at the time of filling is less than the lower limit value of the above range, the gas of fine bubbles in the liquid composition (X1) easily floats before gelling in the step (3), There is concern that the content will decrease. When the viscosity of the liquid composition (X1) at the time of filling exceeds the upper limit of the above range, it becomes difficult to transfer the liquid composition (X1) and it becomes difficult to fill the filling container.
On the other hand, the temperature of the liquid composition (X1) at the time of filling may be any temperature as long as it can achieve the viscosity range of the liquid composition (X1) described above, and may be appropriately selected. Generally, a temperature higher by 5 to 20° C. than the gelation temperature of the liquid composition (X1) may be regarded as a temperature range in which the above-mentioned viscosity range can be achieved.

The filling container is preferably made of a material that does not easily permeate gas so as to suppress gas permeation loss. Examples of the filling container include an aluminum pouch, a bag made of a film having low gas permeability, and a metal container.

As a filling method, a method of filling the liquid composition (X1) in a filling container so that there is no gas phase space as much as possible and quickly sealing it, from the viewpoint of suppressing volatilization loss of gas in a bubble state into the gas phase Is preferred.
The sealing method depends on the type of the filling container, but examples thereof include known sealing methods such as heat sealing and sealing with a lid having an inner lid.

(Process (3))
In a filling container, a liquid composition (X1) containing a gas in a bubble state is cooled to a temperature not higher than the gelation temperature to cause gelation, whereby a bubble state is formed in a substrate composed of the gel composition (X2). A gas-containing material containing the above gas is obtained.

Cooling is preferably performed as quickly as possible. Bubbles contained in the liquid composition (X1) filled in the filling container by rapidly quenching the liquid composition (X1) containing gas in the filling container together with the filling container to a gelling temperature or lower. The volatilization loss of the gas in the state can be reduced as much as possible.

The cooling method is not particularly limited as long as it can quickly cool the filling container filled with the liquid composition (X1) containing the gas in the bubble state to the gelation temperature or lower. As a cooling method, for example, a filling container filled with a liquid composition (X1) containing a gas in a bubble state is rapidly immersed in a water bath cooled to a gelation temperature or lower to quench the filling container together with the filling container. A method in which the filling container is cooled in advance in a refrigerant bath, and the liquid composition (X1) containing gas in a bubble state is filled therein and hermetically sealed.

<Contents>
The content 200 stored in the content storage chamber 104 of the container 1 with a gas supply function differs depending on the use of the container 1 with a gas supply function.
In the case of a cell culture container and a container for transportation, examples of the content 200 include cells, organs, culture media (culture liquid, etc.), and combinations thereof.
In the case of a storage container, the contents 200 include foods, cosmetics, etc. in addition to the contents of the cell culture container and the container for transportation.

<Assembly of container with gas supply function>
By attaching the upper container 30 to the lower container 40 accommodating the gas supply body 50 and attaching the lid 20 to the upper container 30 accommodating the content 200, the partition 12 defines the internal space as the gas supply chamber 102 and the content. The container 1 with a gas supply function is obtained, which includes the container body 10 divided into the object storage chamber 104 and the gas supply body 50 housed in the gas supply chamber 102 of the container body 10.

By attaching an additional upper container 30 below the lower container 40, and by attaching an additional lower container 40 below the additional upper container 30, and repeating this as necessary, the gas supply chamber 102 and the storage of contents are stored. A container with a multistage gas supply function having chambers 104 alternately may be used.

<Mechanism of action>
1. In the container 1 with the gas supply function described above, the container main body 10 and the gas supply body 50 housed in the container main body 10 are provided, and the gas supply body 50 has a gas-containing material, so that gas is supplied. There is no need to separately prepare a high-pressure gas cylinder or the like to do so. Therefore, the structure is simple, compact, and easy to handle. In addition, since the gas-containing material physically holds gas in a bubble state, unlike a conventional atmosphere modifier using a chemical reaction, heat and by-products are not generated when the gas is released. Therefore, the contents or the condition is not deteriorated by heat or by-products.
2. Further, the container main body 10 divides the internal space of the container main body 10 into a gas supply chamber 102 in which the gas supply body 50 is stored and a content storage chamber 104 in which the content 200 is stored. Since the portion 12 is provided, the gas supply body 50 and the content 200 do not come into contact with each other, and the components other than the gas contained in the gas supply body 50 do not easily migrate to the content 200.
3. Further, since the partition part 12 has the gas permeable film 14 facing the content storage chamber 104 side, the gas released from the gas supply body 50 is in contact with the gas permeable film 14 after passing through the gas permeable film 14. Since the gas is forced to pass through the contents 200, the gas can be reliably supplied to the contents 200.
4. Further, since the partition portion 12 has the gas permeable support layer 16 that supports the gas permeable membrane 14, the content 200 can be reliably supported together with the gas permeable membrane 14.

<Other Embodiments>
The container with a gas supply function of the present invention includes a container body and a gas supplier housed in the container body, and the gas supplier may have a specific gas-containing material. It is not limited to the functional container 1.

FIG. 3 is a cross-sectional view showing another example of the container with a gas supply function of the present invention.
The container 2 with a gas supply function includes a container body 60; a gas supply body 50 housed in the container body 60; and a container 70 for contents contained in the container body 60.
The container body 60 has a lid body 20; and a bottomed cylindrical main vessel 62 to which the lid body 20 is attached by screwing. On the upper part of the outer peripheral wall of the main container 62, a male screw portion 66 that is screwed into the female screw portion 28 of the lid 20 is provided.
The contents container 70 has a bottomed cylindrical main container 72 that stores the contents 200; and an isolation film 74 that seals the open end of the main container 72.
The isolation film 74 is a film that is permeable to gas and impermeable to the content 200. Examples of the isolation film 74 include paper, non-woven fabric, microporous film, microporous film, perforated resin film and the like.

FIG. 4 is a cross-sectional view showing another example of the container with a gas supply function of the present invention.
The container 3 with a gas supply function includes a container body 80 formed of a bag body; a gas supply body 50 housed in the container body 80; and a content container 70 housed in the container body 80.
The container body 80 is a bag body made by heat-sealing two sides of two stacked films. A film having low gas permeability is preferable.

FIG. 5: is sectional drawing which shows the other example of the container with a gas supply function of this invention.
The container 4 with a gas supply function includes a container body 80 having a bag body, in which the contents 200 are housed, and a gas supply body 50, which is housed in the container body 80 and on which the contents 200 are placed. Prepare
The container body 80 is a bag body made by heat-sealing two sides of two stacked films. A film having low gas permeability is preferable.
The gas supplier 50 has a gas-containing material 52 and a gas-permeable film 54 that covers the surface of the gas-containing material 52.
In order to prevent the contents 200 from being deteriorated by the oxygen remaining in the container body 80, the container 4 with a gas supply function is provided with a container as shown in FIG. 6 before heat-sealing the container body 80. It is preferable to suck and degas the inside of the main body 80 in advance to reduce the space portion as much as possible.

<Use>
The container with a gas supply function of the present invention can be used as a cell culture container, a transport container, a storage container and the like.

(Cell culture container)
As the cell culture container, a container having a gas permeable membrane in a partition part, such as the container 1 with a gas supply function in FIG. 1, is suitable.
Since the partition 12 of the container 1 with the gas supply function has the gas permeable film 14 facing the content storage chamber 104 side, after the gas released from the gas supply body 50 permeates the gas permeable film 14, Since the gas is forcedly passed through the content 200 in contact with the gas permeable film 14, the gas can be reliably supplied to the content 200. Therefore, the cells can be efficiently cultured.

On the other hand, in the container 2 with the gas supply function of FIG. 3 and the container 3 with the gas supply function of FIG. 4, the content 200 is only exposed to the gas atmosphere in the container body, and compared to the container 1 with the gas supply function. The amount of gas that can be supplied to the contents 200 is small.

(Transportation container, Storage container)
As the container for transportation or the container for storage, any of the containers with a gas supply function such as the container with a gas supply function 1, the container with a gas supply function 2, the container with a gas supply function 3 and the container with a gas supply function 4 is used. It can be preferably used.

(Cell culture method)
The cell culture using the container with a gas supply function of the present invention can be performed in the same manner as the known method except that the known cell culture container is changed to the cell culture container of the present invention.
When culturing cells using the container 1 with a gas supply function, the gas of the content storage chamber 104 is loosened by loosely screwing the lid 20 and the upper container 30 or by removing the lid 20. Is preferably released to the outside. By allowing the gas in the content storage chamber 104 to be released to the outside, the pressure P in the gas supply chamber 102 where the gas is continuously generated from the gas supply body 50 and the pressure P′ in the content storage chamber 104 are The relationship is P>P'. Therefore, the gas released from the gas supply body 50 easily permeates the gas permeable film 14, and the gas can be more reliably supplied to the content 200.

(Content transportation method, content storage method)
The transportation or storage of the contents using the container with a gas supply function of the present invention can be carried out in the same manner as a known method except that the atmosphere regulator is changed to the specific gas supplier of the present invention. In addition, in order to prevent the contents from being deteriorated by the oxygen remaining in the container body, the inside of the container body may be replaced with an inert gas such as nitrogen before sealing the container body.

Hereinafter, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto.

<Measurement>
The methods for measuring each physical property in the examples are shown below.

(Gelization temperature)
The gelling temperature was determined as follows.
A 50 mL glass screw bottle was charged with the liquid composition (X1) of about half the capacity of the screw bottle and the container was tightly sealed, immersed in a constant temperature water bath, heated to 75° C., and then gradually cooled down in the constant temperature water bath. The temperature at which the liquid composition stopped flowing even when the screw bottle was tilted at 45° and 90° was defined as the gelling temperature. The temperature was measured by attaching a standard mercury thermometer to the outer wall of the screw bottle.

(viscosity)
The viscosity was measured using an E-type viscometer (manufactured by Toki Sangyo Co., Ltd., for low viscosity: RE-105L, for high viscosity: RE-215U).

(Observation of dispersed state of bubbles in gas-containing material)
The gas containing material was cut with a sharp knife. The dispersed state of the bubbles was observed by observing the cross section using a microscope (Digital Microscope VHX-900F, manufactured by Keyence Corporation).

(Content of hydrogen gas in the gas-containing material)
A sample collected from the gas-containing material under conditions of atmospheric pressure and 10° C. was precisely weighed and sealed in a headspace GC analysis sample bottle used in gas chromatographic analysis (hereinafter referred to as GC analysis). The sample bottle was placed in a constant temperature water bath at 70° C., and the sample was heated until it became a liquid composition (X1) containing no bubbles, and hydrogen gas was released into the gas phase. The gas phase gas in the sample bottle is sampled, the hydrogen gas is quantified by GC analysis (TCD detector), and the hydrogen gas content in the gas-containing material (volume/mass% of gas-containing material (v/w %) conversion) was calculated.

(Contents of nitrogen gas, oxygen gas, and argon gas in the gas-containing material)
The content of each gas in the gas-containing material (volume/mass% (v/w%) conversion of the gas-containing material) is calculated by the same method as the measuring method of "content of hydrogen gas in gas-containing material". did.

(Propane gas content in gas-containing material)
The TCD detector for GC analysis was changed to an FID detector, and the content of propane gas in the gas-containing material (gas-containing material was measured by the same method as the method for measuring “content of hydrogen gas in gas-containing material”). Volume/mass% (v/w %) conversion) was calculated.

(Content of carbon dioxide gas in the gas-containing material)
A sample collected from the gas-containing material under atmospheric pressure and 10° C. was precisely weighed and sealed in a headspace TOC analysis sample bottle used in total organic carbon analysis (hereinafter referred to as TOC analysis). The sample bottle was placed in a constant temperature water bath at 70° C., and the sample was heated until it became a liquid composition (X1) containing no bubbles, and carbon dioxide gas was released into the gas phase. The gas phase gas in the sample bottle was sampled, and the carbon dioxide gas released into the gas phase was quantified by TOC analysis. On the other hand, in order to quantify the carbon dioxide gas remaining in the liquid composition (X1) after heating at 70° C., a 0.1 N potassium hydroxide aqueous solution was placed in a sample bottle after collecting the gas phase gas. Was added to make the sample basic, and the sample bottle was shaken to dissolve the total amount of carbon dioxide in the bottle as an alkali salt in the liquid. The total amount of carbon dioxide in the liquid composition (X1) is determined by TOC analysis of this dissolved solution, and the content of carbon dioxide gas in the gas-containing material (volume/mass% of gas-containing material (v/w% ) Conversion) was calculated.

(Time-dependent change of gas content in gas-containing material in gas-tight container at 25°C (gas release characteristic))
A sample collected from the above-mentioned various gas-containing materials under atmospheric pressure and 10° C. is precisely weighed in a range of 0.8 to 1.5 g in a headspace GC analysis sample bottle (21.75 mL) used for GC analysis. And sealed. This sample bottle was allowed to stand in a thermostat at 25° C., gas phase gas in the sample bottle was sampled every time, and it was released into the gas phase from the gas-containing material at 25° C. by GC analysis. Time variation of the content of various gases in the gas-containing material (volume/mass% (v/w%) conversion of the gas-containing material) by quantifying the amount of gas and subtracting it from the gas content when producing the gas-containing material I asked.

<<Example 1>>
<Manufacture of gas-containing material>
(Preparation of liquid composition (X1))
A 1 L resin container with a lid was charged with fish scale-derived gelatin powder and tap water (Kitakyushu City Waterworks Bureau) at a gelatin concentration of 35% by mass, and allowed to stand at room temperature to swell gelatin. A liquid composition (X1) (gelatin aqueous solution) was prepared by immersing the container in a warm water bath at 70° C. to dissolve gelatin.
The gelling temperature of the liquid composition (X1) was 26°C.

(Process (1))
A 1 L poly container was charged with 500 g of the liquid composition (X1) (gelatin aqueous solution), immersed in a constant temperature water bath at 70° C., and heated to hold. Hydrogen gas was bubbled through the bottom of the plastic container at 100 mL/min for about 15 minutes. A liquid container (X1) containing hydrogen gas in a bubble state was obtained by sealing the plastic container, taking it out from the constant temperature water tank, and rapidly shaking it by hand until the internal temperature reached 30 to 35°C. It was

(Process (2))
The liquid composition (X1) containing hydrogen gas in a bubble state was immediately filled in an aluminum pouch with an internal temperature of 30 to 35° C. so that there was no vapor phase space, and then sealed.

(Process (3))
An aluminum pouch filled with a liquid composition (X1) containing hydrogen gas in a bubble state is immediately immersed in water at 5° C., and the liquid composition (X1) is cooled to gel to give a gel composition. A gas-containing material was obtained in which hydrogen gas in a bubble state was included in the base material composed of the substance (X2).

When the content of hydrogen gas in the obtained gas-containing material was measured by GC analysis, it was 28% by volume in terms of volume/mass% (v/w%) of the gas-containing material.
FIG. 7 shows a photograph of the appearance of the liquid composition (X1) (aqueous gelatin solution) and the gas-containing material at 10° C. before supplying hydrogen gas. Although it is transparent before supplying hydrogen gas, it can be seen that the gas-containing material in which fine gas is dispersed becomes white and opaque.
A photograph of a cross-sectional microscope observation of the gas-containing material is shown in FIG. It was confirmed that fine bubbles of 200 μm or less were dispersed at a high concentration.
Further, FIG. 15 shows the time change of the content of hydrogen gas in the gas-containing material in the closed container at 25° C. The content of hydrogen gas was reduced to 14% by volume after 1 hour and 8% by volume after 5 hours, and it was confirmed that hydrogen gas was released in the gas phase.

<<Example 2>>
<Gas release test from gas-containing material using container with gas supply function>
In the gas supply chamber 102 (capacity: 14.5 cm ³ ) of the container 1 with a gas supply function shown in FIG. 2.5 g of ultrapure water was stored in the content storage chamber 104. The lid 20 was removed from the upper container 30, and the container 1 with the gas supply function was left in an incubator maintained at 37°C. The gas phase gas in the gas supply chamber 102 was sampled every predetermined time, and the hydrogen gas concentration in the gas phase was measured by GC analysis. FIG. 9 shows the time change of the hydrogen gas concentration in the gas phase. It has been confirmed that the gas supply chamber 102 can be maintained in a hydrogen gas atmosphere for several hours by releasing hydrogen gas from the hydrogen gas-containing material.

<<Example 3>>
<Manufacture of gas-containing material>
The same operation as in Example 1 was performed, except that the hydrogen gas was changed to the oxygen gas, to obtain a gas-containing material containing oxygen gas in a bubble state.
When the content of oxygen gas in the obtained gas-containing material was measured by the GC analysis method, it was 32 volume% in terms of volume/mass% (v/w%) of the gas-containing material.
FIG. 7 shows a photograph of the appearance of the gas-containing material at 10° C. It can be seen that the gas-containing material in which fine oxygen gas is dispersed becomes white and opaque.
A photograph of a cross-sectional microscope observation of the gas-containing material is shown in FIG. It was confirmed that fine bubbles of 200 μm or less were dispersed at a high concentration.
Further, FIG. 15 shows the time change of the oxygen gas content in the gas-containing material in the closed container at 25° C. The content of oxygen gas was reduced to 12% by volume after 3 hours and to 8% by volume after 5 hours, confirming that oxygen gas was released in the gas phase.

<<Example 4>>
<Manufacture of gas-containing material>
The same operation as in Example 1 was carried out except that the hydrogen gas was changed to nitrogen gas to obtain a gas-containing material containing nitrogen gas in a bubble state.
When the content of nitrogen gas in the obtained gas-containing material was measured by GC analysis, it was 46% by volume in terms of volume/mass% (v/w%) of the gas-containing material.
FIG. 7 shows a photograph of the appearance of the gas-containing material at 10° C. It can be seen that the gas-containing material in which fine nitrogen gas is dispersed becomes white and opaque.
A photograph of a cross-sectional microscope observation of the gas-containing material is shown in FIG. It was confirmed that fine bubbles of 200 μm or less were dispersed at a high concentration.
Further, FIG. 15 shows the time change of the nitrogen gas content in the gas-containing material in the closed container at 25° C. The content of nitrogen gas was reduced to 27% by volume after 1 hour, 21% by volume after 3 hours, and 17% by volume after 5 hours, and it was confirmed that nitrogen gas was released in the gas phase.

<<Example 5>>
<Manufacture of gas-containing material>
The same operation as in Example 1 was carried out except that the hydrogen gas was changed to argon gas, to obtain a gas-containing material containing argon gas in a bubble state.
When the content of argon gas in the obtained gas-containing material was measured by the GC analysis method, it was 28 volume% in terms of volume/mass% (v/w%) of the gas-containing material.
FIG. 7 shows a photograph of the appearance of the gas-containing material at 10° C. It can be seen that the gas-containing material in which fine argon gas is dispersed becomes white and opaque.
A photograph of a cross-sectional microscope observation of the gas-containing material is shown in FIG. It was confirmed that fine bubbles of 200 μm or less were dispersed at a high concentration.
Further, FIG. 15 shows a time change of the content of the argon gas in the gas-containing material in the closed container at 25° C. The content of argon gas was reduced to 19% by volume after 1 hour, 11% by volume after 3 hours, and 8% by volume after 5 hours, and it was confirmed that the argon gas was released in the gas phase.

<<Example 6>>
<Manufacture of gas-containing material>
The same operation as in Example 1 was carried out except that the hydrogen gas was changed to propane gas to obtain a gas-containing material containing propane gas in a bubble state.
When the content of propane gas in the obtained gas-containing material was measured by the GC analysis method, it was 27 volume% in terms of volume/mass% (v/w%) of the gas-containing material.
A photograph of a cross-sectional microscope observation of the gas-containing material is shown in FIG. It was confirmed that fine bubbles of 200 μm or less were dispersed at a high concentration.
In addition, FIG. 15 shows the time change of the content of propane gas in the gas-containing material in the closed container at 25° C. The content of propane gas was reduced to 18% by volume after 1 hour, 14% by volume after 3 hours, and 11% by volume after 5 hours, and it was confirmed that propane gas was released in the gas phase.

<<Example 7>>
<Manufacture of gas-containing material>
The same operation as in Example 1 was performed except that the hydrogen gas was changed to carbon dioxide gas to obtain a gas-containing material containing carbon dioxide gas in a bubble state.
When the gas content of carbon dioxide in the obtained gas-containing material was measured by the GC analysis method, it was 72 volume% in terms of volume/mass% (v/w%) of the gas-containing material.
FIG. 7 shows a photograph of the appearance of the gas-containing material at 10° C. It can be seen that the gas-containing material in which fine carbon dioxide gas is dispersed becomes white and opaque.
A photograph of a cross-sectional microscope observation of the gas-containing material is shown in FIG. It was confirmed that fine bubbles of 200 μm or less were dispersed at a high concentration.
Further, FIG. 15 shows the time change of the content of carbon dioxide gas in the gas-containing material in the closed container at 25°C. The content of carbon dioxide gas was reduced to 9% by volume after 1 hour and 3% by volume after 3 hours, and it was confirmed that carbon dioxide gas was released in the gas phase.

INDUSTRIAL APPLICABILITY The container with a gas supply function of the present invention is useful as a cell culture container, a transportation container, a storage container and the like.

1 container with gas supply function,
2 container with gas supply function,
3 container with gas supply function,
4 container with gas supply function,
10 container body,
12 partitions,
14 gas permeable membrane,
16 support layer,
18 through holes,
20 lid,
22 tubular part,
24 canopy,
28 Female thread,
30 upper container,
32 tubular part,
36 Upper male thread,
38 Lower female thread,
40 Lower container,
42 tubular part,
44 bottom,
46 upper male thread,
48 lower female thread,
50 gas supplier,
52 gas-containing material,
54 gas permeable membrane,
60 container body,
62 main container,
66 Male thread,
70 Containers for contents,
72 Main container,
74 isolation membrane,
80 container body,
102 gas supply chamber,
104 contents storage room,
200 contents.

Claims (15)

A container body and a gas supply body housed in the container body ,
The container body has a gas permeable partitioning portion that divides the internal space of the container body into a gas supply chamber in which the gas supply body is stored and a content storage chamber in which contents are stored,
A container with a gas supply function, wherein the gas supplier has the following gas-containing material.
Gas-containing material: In a substrate comprising either one or both of a liquid composition containing a gelling agent and a liquid medium, and a gel composition containing a network structure formed from the gelling agent and a liquid medium. Including gas in a bubble state. The container with a gas supply function according to claim 1, wherein the partition section divides the internal space into upper and lower parts. The container with a gas supply function according to claim 1, wherein the content storage chamber has a lid. The container with a gas supply function according to any one of claims 1 to 3, wherein the partition part has a gas permeable film facing the content storage chamber side. The container with a gas supply function according to any one of claims 1 to 4, wherein the partition part has a gas-permeable support layer that supports the gas-permeable membrane. Said gas, hydrogen, oxygen, nitrogen, argon, helium, carbon dioxide, methane, ethane, comprising at least one gas selected from the group consisting of propane and butane, according to any one of claims 1 to 5 Container with gas supply function. The content of the gas in the gas-containing material is 6 to 80 volume% in terms of volume/mass% (v/w%) of the gas-containing material, according to any one of claims 1 to 7. Container with gas supply function. The bubble size of the gas bubbles states encompassed gas-containing material is, microbubbles size or less, the gas supply function container according to any one of claims 1-7. The gelling agent is gelatin, is at least one of agar, carrageenan, selected pectin, glucomannan, pullulan, sodium alginate, and carboxyvinyl polymers or the group consisting of a derivative thereof, any one of the claims 1-8 A container with a gas supply function according to the item. Made from the gas supply function container according to any one of claims 1 to 9 cell culture vessel. Made from the gas supply function container according to any one of claims 1 to 9 carrying container. Made from the gas supply function container according to any one of claims 1 to 9 storage container. Culturing cells using the gas supply function container according to any one of claims 1 to 9 cell culture methods. Carrying the contents by using the gas supply function container according to any one of claims 1 to 9 contents delivery method. To save the contents by using the gas supply function container according to any one of claims 1 to 9 content storage method.