WO2012099249A1 - Procédé de stockage à basse température et récipient de stockage à basse température - Google Patents

Procédé de stockage à basse température et récipient de stockage à basse température Download PDF

Info

Publication number
WO2012099249A1
WO2012099249A1 PCT/JP2012/051244 JP2012051244W WO2012099249A1 WO 2012099249 A1 WO2012099249 A1 WO 2012099249A1 JP 2012051244 W JP2012051244 W JP 2012051244W WO 2012099249 A1 WO2012099249 A1 WO 2012099249A1
Authority
WO
WIPO (PCT)
Prior art keywords
liquid
container
storage
temperature
pressure
Prior art date
Application number
PCT/JP2012/051244
Other languages
English (en)
Japanese (ja)
Inventor
木野　正人
昭夫 清水
Original Assignee
株式会社ミツヤコーポレーション
学校法人創価大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 株式会社ミツヤコーポレーション, 学校法人創価大学 filed Critical 株式会社ミツヤコーポレーション
Priority to CN201280006028.5A priority Critical patent/CN103327839B/zh
Priority to JP2012553787A priority patent/JP5958913B2/ja
Publication of WO2012099249A1 publication Critical patent/WO2012099249A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M45/00—Means for pre-treatment of biological substances
    • C12M45/22—Means for packing or storing viable microorganisms
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0236—Mechanical aspects
    • A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
    • A01N1/0252—Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/36—Freezing; Subsequent thawing; Cooling
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/36—Freezing; Subsequent thawing; Cooling
    • A23L3/363—Freezing; Subsequent thawing; Cooling the materials not being transported through or in the apparatus with or without shaping, e.g. in form of powder, granules, or flakes
    • A23L3/364—Freezing; Subsequent thawing; Cooling the materials not being transported through or in the apparatus with or without shaping, e.g. in form of powder, granules, or flakes with packages or with shaping in form of blocks or portions

Definitions

  • the present invention relates to a cryopreservation method for preserving preservation objects such as foods, organs and cells at low temperatures, and a cryopreservation container used in this cryopreservation method.
  • organic substances such as foods, organs and blood can suppress the activity and chemical reaction of bacteria and enzymes, and the storage time becomes longer as the storage temperature is lower. Therefore, when storing an object to be stored for a long time, it is often stored frozen by freezing, amorphous solidification, or the like.
  • the freezing point of water decreases by about 1 ° C. every time the pressure increases by 10 MPa up to about 200 MPa. That is, in order to decrease the storage temperature in the non-freezing state by 1 ° C., the pressure may be increased by about 10 MPa.
  • a pressure freezing is performed in which a high pressure of about 200 MPa is applied to the pressure vessel using a pressure device, and the object to be stored is kept in a non-freezing state at about ⁇ 20 ° C.
  • a law has been proposed.
  • the object to be stored may be frozen by reducing the pressure to normal pressure.
  • a high pressure of 400 to 500 MPa may be applied using a pressurizing device to denature proteins and enzymes such as foods and may be stored in a non-frozen state at a low temperature.
  • the conventional pressure freezing method requires a large-scale pressure device that generates high pressure.
  • Patent Document 1 food and storage water are put in a sealed container (pressure container) to freeze part of the stored water, and the pressure in the sealed container is increased from atmospheric pressure by volume expansion of the generated ice.
  • a technique for storing food at a low temperature in a non-freezing state at a temperature below the freezing point at normal pressure is disclosed.
  • Non-Patent Document 1 discloses that in a non-freezing state, water or an aqueous solution is filled in a pressure-resistant container containing food, frozen at a sub-freezing temperature, and high pressure is applied in the pressure-resistant container. A technique for impregnating food with seasoning water is disclosed.
  • the conventional freeze pressurization method requires an expensive pressure vessel. And, if the amount of ice exceeding the expected amount is generated, such as when the temperature in the pressure vessel is too low, a high pressure is suddenly applied in the pressure vessel, so the pressure vessel may be damaged, An expensive pressure vessel needs to be repaired or remanufactured. Moreover, the pressure vessel is heavy and difficult to carry.
  • the pressure vessel may be damaged.
  • the present invention provides a low-temperature storage method in which an excessively high pressure is not applied, and a low-temperature storage container suitable for this low-temperature storage method, while relieving abrupt changes in the internal pressure of the container containing the storage object.
  • the purpose is to provide.
  • the cryopreservation method of the present invention has a stretchable first closed space filled with a first liquid that increases in volume when solidified, a compressibility greater than that of the first liquid, and is higher than that of the first liquid.
  • Forming a retractable second closed space filled with a second liquid having a low-temperature freezing point in the container, placing a storage object in the container, and first and second closures A step of sealing the container in which the space is formed and the object to be stored is sealed; and after sealing the container, the first liquid and the second liquid are cooled to a temperature lower than the freezing point of the first liquid. Storing the storage object at a storage temperature lower than the freezing point of the liquid in the storage object at the atmospheric pressure, the temperature being between the freezing point of the liquid and the liquid.
  • the low-temperature storage method of the present invention after the container is sealed, when the first liquid is cooled to a temperature lower than the freezing point of the first liquid, at least a part of the first liquid is solidified (frozen). Since the first closed space occupied by the first liquid expands due to the increase in volume when the first liquid solidifies, the volume in the container other than the first closed space decreases, so that the pressure in the container is reduced. Ascend and exceed atmospheric pressure.
  • the second liquid filled in the second closed space has a higher compressibility than the first liquid, a part of the expanded volume of the first closed space is equal to the volume of the second closed space. Absorbed by reduction. And since the freezing point of the second liquid is lower than the storage temperature, it does not freeze even when cooled to the storage temperature. Therefore, compared with the case where the second closed space filled with the second liquid is not formed in the container, the rapid increase in pressure in the container is suppressed and the pressure increase becomes slow.
  • the upper limit value of the pressure in the container and the pressure increase mode according to the upper limit of the internal volume of the container, the amount of the first liquid, the amount of the second liquid, and the cooling mode. is uniquely determined. Therefore, there is no possibility that the container will be damaged due to an increase in pressure more than expected, and the object to be stored can be stably stored at a low temperature in a non-frozen state.
  • Non-Patent Document 1 In the conventional freeze pressurization method, for example, as disclosed in Non-Patent Document 1 described above, a seasoning liquid is filled in a pressure-resistant container containing a food surrounded by a seasoning liquid, and the aqueous solution is frozen at ⁇ 25 ° C. A high pressure of several hundred MPa is applied. However, in this method, the food is frozen only by a slight change in the ratio between the aqueous solution concentration and the seasoning solution concentration.
  • a desired high pressure is obtained by adjusting the amount of liquid, and a desired high pressure can be stably obtained as compared with the conventional method.
  • the freezing point of the first liquid is lowered by pressurization, and a drug such as a freezing point lowering agent for lowering the freezing point is not required.
  • a chemical such as a freezing point reducing agent may be added to the first liquid.
  • the object to be preserved may be contained in a space other than the first and second closed spaces in the container, and the space may be filled with a liquid having a freezing point lower than the preservation temperature. .
  • the second closed space is included in the first closed space
  • the object to be stored is included in the second closed space
  • the temperature other than the first closed space in the container is lower than the storage temperature.
  • a liquid having a freezing point of may be filled.
  • the internal space in the container is the first closed space
  • the second closed space may be included in the first closed space
  • the storage object may be included in the second closed space.
  • the second closed space is formed in the first closed space, and the storage object is stored in the container.
  • the object to be stored is in the second enclosed space, and in the step of increasing the pressure in the container, at least a part of the first liquid is frozen to produce an icing capsule, It is preferable to increase the pressure in the closed space of 2 to a pressure exceeding the atmospheric pressure.
  • the object to be stored is contained in the first closed space filled with the first liquid. Therefore, an object to be stored is contained in an icing capsule formed by freezing the first liquid. Therefore, the storage object can be stored at a low temperature in the frozen capsule.
  • the second liquid serves as a buffer material, and a rapid pressure increase in the frozen capsule is suppressed. Therefore, since cracks do not occur in the frozen capsule, the pressure in the frozen capsule does not decrease, and the storage object can be stably stored at a low temperature in a non-frozen state.
  • the first liquid and the first liquid it is preferable to set the volume ratio with respect to the second liquid.
  • the method further includes a step of forming a third closed space filled with a liquid or a gas to be contained in the second closed space.
  • the storage object contacts only the liquid or gas filled in the third closed space and does not contact the second liquid. Therefore, even if the storage object is a food, organ, cell, or the like, the liquid that denatures the storage object can be filled in the second space, and the options for the second liquid are widened.
  • the first closed space is filled with water as the first liquid in the step of forming the first and second closed spaces in the container.
  • the first liquid may be fresh water or water added with a chemical.
  • the freezing point reducing agent is added, the freezing temperature is lowered. Therefore, in the step of forming the first and second closed spaces in the container, the first liquid to which the freezing point reducing agent is added is used as the first closed space. It is preferable to fill the inside.
  • silicone oil, vegetable oil, gelled Yamanori or gelatin is used as the second liquid as the second liquid. It is preferable to fill the space.
  • oils have a large compressibility, and particularly silicone oil has a large compressibility. Therefore, it is preferable to fill the second closed space with silicone oil.
  • the second liquid is vegetable oil having a low freezing point, or gelled Yamatonori or gelatin.
  • cryopreservation method of the present invention is not limited to the case where it is applied to pressurized non-freezing cryopreservation in which the storage object is cryopreserved in an unfrozen state, and the preservation object is frozen in a pressurized state to be cryopreserved.
  • Pressurized freezing and cryogenic storage pressurize the object to be stored in a non-frozen state by pressurization, and then reduce the pressure to form a uniform ice nucleus in the object to be stored to freeze the object to be stored. It can also be applied to laws.
  • the cryopreservation container of the present invention is a container filled with a first liquid that increases in volume when solidified, is contained in the container, has a higher compressibility than the first liquid, and is lower in temperature than the first liquid. Filled with a second liquid having a freezing point and made of a stretchable inner bag, the temperature between the freezing point of the first liquid and the second liquid, and the atmospheric pressure of the liquid in the storage object The storage object is stored at a storage temperature lower than the freezing point.
  • the cryopreservation container of the present invention is suitably used in the cryopreservation method of the present invention.
  • Explanatory drawing which shows the state which concerns on the 1st Embodiment of this invention and cools a cryopreservation container with an iced capsule formation tank.
  • Explanatory drawing which shows a cryopreservation container.
  • Explanatory drawing which shows the cryogenic storage container which concerns on the 2nd Embodiment of this invention.
  • the cryopreservation container 10 containing the storage object A is suspended in the contact solvent 4 accommodated in the frozen capsule formation tank 3 by the hanging rod 2 via the hanger 1.
  • the ice capsule forming tank 3 contains a contact refrigerant 4 such as ethanol brine, and is placed in a freezer compartment (not shown) and cooled.
  • the storage object A is suspended in the contact refrigerant 4 while being stored in the cryogenic storage container 10 shown in FIG.
  • the cryopreservation container 10 has a triple structure, and includes an outer container 11 (container), an inner bag 12, an inner bag 13, and a weight 14, and the upper portion of the outer container 11 is a hanger. 1 is fixed.
  • the outer container 11 has an upper limit on the internal volume, and is, for example, a bag made of a resin such as a polyethylene film.
  • the inner bag 12 and the inner bag 13 are extendable and made of, for example, a resin such as polyethylene. It is preferable that the inner bag 12 and the inner bag 13 are not in contact with the inner surface of the outer container 11 and the inner surface of the inner bag 12 with a spacer (not shown), respectively.
  • the inside of the inner bag 13 contains the storage object A and is filled with liquid or gas B.
  • the storage object A is, for example, food such as vegetables, fruits, meat, fish, human or animal organs, cells, etc., and the size and thickness thereof are not particularly limited.
  • the liquid or gas B filled in the inner bag 13 is not particularly limited, but preferably does not denature the storage object A.
  • the liquid or gas B may be a seasoning liquid such as saline or carbon dioxide gas when the storage object A is a food, or a low-temperature storage liquid when the storage object A is an organ or a cell. Also good.
  • the inner bag 13 contains the inner bag 13, and the internal space other than the space occupied by the inner bag 4 of the inner bag 12 has a higher compression rate than water (first liquid) and a freezing point lower than that of water. It is filled with the liquid C (2nd liquid) which has.
  • Liquid C is, for example, oils, gelled Yamatonori or gelatin.
  • oils have a large compressibility
  • silicone oil in particular has a large compressibility.
  • dimethyl silicone oil has a large compressibility. Therefore, when the storage temperature is ⁇ 5 ° C. or lower, the liquid C is preferably a silicone oil, particularly dimethyl silicone oil.
  • the storage object A is a food
  • the liquid C is an edible low-freezing point vegetable oil or a gelled Yamatonori
  • the food safety due to liquid leakage can be ensured.
  • corn oil, coconut oil, and walnut oil have a freezing point at normal pressure of less than 0 ° C., and are suitable when the storage temperature is a relatively high temperature of ⁇ 2 ° C. to ⁇ 5 ° C.
  • the outer container 11 contains the inner bag 12 and the inner space other than the space occupied by the inner bag 12 of the outer container 11 is filled with the liquid D (first liquid) that increases in volume when solidified.
  • the liquid D is preferably water.
  • the liquid D may be fresh water or a solution obtained by adding a drug such as a freezing point lowering agent (freezing inhibitor) to fresh water.
  • the space in the outer container 11 corresponds to the first closed space in the present invention
  • the space in the inner bag 12 corresponds to the second closed space in the present invention
  • the space in 13 corresponds to the third closed space in the present invention.
  • the cryopreservation method forms in the outer container 11 a stretchable first closed space filled with the liquid D and a stretchable second closed space filled with the liquid C.
  • the cryopreservation container 10 containing the storage object A is suspended in the contact refrigerant 4 of the freeze capsule formation tank 3, and the freeze capsule formation tank 3 is installed in the freezer compartment.
  • the liquid D is cooled to a temperature lower than the freezing point of the liquid D to freeze at least a part of the liquid D, and the internal pressure exceeds atmospheric pressure.
  • the temperature is between the step of freezing at least a part of the liquid D to form a frozen capsule and the freezing point of the liquid C and the liquid D, and the object to be stored Storing the storage object A at a storage temperature lower than the freezing point of the liquid (water) in A at atmospheric pressure.
  • the liquid D (water) in the outer container 11 is cooled to form an ice capsule.
  • the volume of the inner bag 12 decreases due to volume expansion when water freezes, and the pressure in the inner bag 12 rises to atmospheric pressure or higher.
  • the liquid C filled in the inner bag 12 is compressed and its volume is reduced, so that a part of the pressure increase is buffered, and the sudden pressure increase in the inner bag 12 is suppressed, so It is maintained and does not crack. And since the pressure in the inner bag 12 exceeds atmospheric pressure, the freezing temperature of the water in the storage object A is less than 0 ° C., and the storage object A is stored in a non-freezing state at a low temperature of less than 0 ° C. Is possible.
  • the pressure increase becomes slow there is no possibility that the outer container 11 is unintentionally damaged, and it is not always necessary to use a pressure resistant container as the outer container 11.
  • a pressure vessel is used as the outer container 11, the wall thickness is small and the pressure resistance may be low, so that a light and inexpensive pressure vessel can be used.
  • the volume expansion during freezing of the liquid D water
  • the volume ratio between the liquid C and the liquid D may be set in consideration of the pressure that does not freeze.
  • the pressure upper limit value and the pressure increase mode in the outer container 11 are uniquely determined. Therefore, it is possible to stably store the storage object A at a low temperature in a non-freezing state without damaging the outer container 11.
  • a pressure-resistant container containing a food surrounded by a seasoning liquid is filled with an aqueous solution (seasoning liquid) such as salt or sugar and is -25.
  • an aqueous solution seasoning liquid
  • a high pressure of several hundred MPa is applied.
  • the pressure changes when the concentration of the aqueous solution changes slightly, and the food freezes.
  • a desired high pressure is obtained by adjusting the amount of liquid, and a desired high pressure can be stably obtained as compared with the conventional case.
  • This embodiment exhibits a good effect particularly when foods, organs, cells and the like are stored at a low temperature for a relatively short period of time.
  • the freezing capsule is forcibly generated to depressurize the inside and simultaneously take away the latent heat inside. As a result, it is possible to shorten the freezing time and realize high-quality freezing.
  • the sphere has the smallest volume for the same surface area. Therefore, when the outer shape of the inner bag 12 filled with the liquid C (water) is a spherical surface, the effect of absorbing the volume increase due to the freezing of the liquid C is small. However, since the sphere can uniformly disperse a large internal pressure, the outer shape of the inner bag 12 should be close to a spherical surface. Therefore, the outer shape of the inner bag 12 is preferably a spheroid.
  • the elliptical flatness is a flatness close to that of a sphere, and is preferably near the limit where the frozen capsule is not broken by the elasticity of the liquid C.
  • a bag made of polyethylene film having a clamping chuck at the top was used as the outer container 11. This bag had a length under the chuck of 140 mm, a width of 100 mm, and a thickness of 0.04 mm.
  • the inner bag 12 a bag made of a polyethylene film having a fastening chuck on the upper side was used as the inner bag 12. This bag had a length under the chuck of 85 mm, a width of 60 mm, and a thickness of 0.04 mm.
  • Example 1 In Example 1 and Comparative Examples 1 to 3, edible walnut oil was used as liquid C.
  • Example 1 and Comparative Examples 1 to 3 frozen capsules were formed using a volume ratio of 7 for water and 9 for edible walnut oil.
  • the frozen capsules were stored in a refrigerator at low temperatures for 3 days at storage temperatures of ⁇ 3 ° C., ⁇ 7 ° C., ⁇ 18 ° C., and ⁇ 60 ° C.
  • the control temperature range of the storage temperature is 2 ° C. above and below the center temperature.
  • Example 2 and Comparative Examples 4 and 5 the same treatment as in Example 1 and Comparative Examples 1 to 3 was performed using dimethyl silicone oil having a viscosity of 10 mm / s as liquid C.
  • the experimental results are shown in Table 1.
  • liquid C is dimethyl silicone oil
  • the storage temperature at which the frozen capsule does not crack is lower than when the liquid C is edible walnut oil. This is presumably because dimethyl silicone oil has a higher compressibility than edible walnut oil and can absorb more volume expansion due to freezing of liquid D (water).
  • Experiment 2 Next, frozen capsules were formed by the same method and conditions as in Experiment 1, and Experiment 2 was performed at the same low temperature storage conditions as in Experiment 1.
  • 10 ml of 1% saline solution as liquid B, 90 ml of dimethyl silicone oil having a viscosity of 10 mm / s as liquid C, 70 ml of fresh water as liquid D, and 1.5% agar as storage object A were used.
  • the results of Experiment 2 are shown in Table 2.
  • Comparative Example 7 is a control sample at 20 ° C. that is not stored at low temperature.
  • the 1.5% agar used as the storage object A is a sample formed by mixing a Takano tofu powder in a 1.5% by weight agar aqueous solution.
  • the moisture content of this sample is about 80%, which is close to the moisture content of meat such as seafood and animal meat, which is 65% to 85%.
  • This sample was specifically prepared as follows. First, Koya tofu was crushed with a grater and powdered. Then, 7% by weight of Takano tofu powder with respect to the weight of water was mixed with the boiled 1.5% by weight agar aqueous solution, stirred for 5 minutes, placed in the container, and cooled with ice water to solidify the contents. . And after removing the condensed water droplets in the container, die-cutting was performed using a cylindrical mold having an inner diameter of 12 mm to obtain a cylindrical sample having a diameter of 12 mm and a height of 10 mm. Further, the molded sample was stored in a plastic bag with a chuck and refrigerated at 4 ° C. for 1 day.
  • the drip rate R was determined using this sample as follows. First, after measuring the weight Wp of the sample before cryopreservation, the soot sample was cryopreserved. Then, the sample was placed in a centrifuge tube (Spitz tube) in a cryopreserved state, and naturally thawed by centrifuging at 113 G for 5 minutes using a swing rotor centrifuge. And the sample was taken out from the centrifuge tube, the weight Wc was measured, and the drip rate R was calculated
  • Example 7 Although the freezing capsule cracked, the drip rate R was small as compared with Comparative Example 6 without dimethyl silicone oil. This is presumably because in Example 7, the frozen capsules were cracked after deep supercooling once, so that the sample tissue was not damaged much.
  • Example 7 even when cryopreserved in a frozen state, the presence of dimethylsilicone oil takes a long time until ice breaks, and the sample is not frozen due to a large freezing point depression due to pressurization. Since the ice nuclei are uniformly formed inside the sample by freezing due to the ice breakage from the state, the sample is frozen and can be stored at a low temperature of good quality.
  • the cryopreservation container 20 includes an outer container 21, a first inner bag 22, and a second bag 23. Although not shown, the upper portion of the outer container 21 is the hanger 1 (see FIG. 1). Fixed to.
  • the outer container 21 is, for example, a container made of a resin such as polyethylene terephthalate (PET) or a metal such as iron.
  • the first bag 22 and the second inner bag 23 are extendable and made of, for example, a resin such as polyethylene.
  • the first bag 22 is filled with the liquid C having a compression rate larger than that of the liquid D and having a freezing point lower than the freezing point of the liquid D in the first closed space inside.
  • the liquid C filled in the first bag 22 is oils, gelled Yamatonori, gelatin, and the like, similar to the liquid C filled in the inner bag 22.
  • the second bag 23 is filled with the liquid D whose volume increases when solidified in the second closed space inside.
  • the liquid D is water.
  • the liquid D may be fresh water or a solution obtained by adding a drug such as a freezing point lowering agent to fresh water.
  • the outer container 21 contains the first bag 22, the second bag 23, and the storage object A, and the inner space other than the space occupied by these in the outer container 21 is filled with an antifreeze E such as an ethanol aqueous solution.
  • an antifreeze E such as an ethanol aqueous solution.
  • the storage object A is, for example, food such as vegetables, fruits, meat, fish, human or animal organs, cells, and the like, and the size and thickness are not particularly limited, but are preferably smaller.
  • cryogenic storage container 20 containing the storage object A is cooled to a storage temperature in a freezing chamber (not shown).
  • the liquid D (water) in the second bag 23 is cooled and frozen. Due to the volume expansion during freezing, the volume of the liquid portion in the outer container 21 decreases, and the pressure in the outer container 21 rises above atmospheric pressure.
  • the outer container 21 is not likely to be inadvertently damaged by an excessively high pressure, and the wall thickness is thin and the pressure resistance may be low. Therefore, a lightweight and inexpensive pressure container can be used. .
  • the volume expansion during freezing of the liquid D water
  • the compressibility of the liquid C and the moisture in the storage object A at the storage temperature.
  • the volume ratio between the liquid C and the liquid D may be set in consideration of the pressure at which the liquid does not freeze.
  • the upper limit value and the rising mode of the pressure applied in the outer container 21 are uniquely determined. Therefore, it is possible to stably store the storage object A at a low temperature in a non-freezing state without damaging the outer container 21.
  • the present embodiment obtains a desired high pressure by adjusting the amount of liquid, and can stably obtain a desired high pressure as compared with the conventional case.
  • cryopreservation containers 10 and 20 are suspended and cooled in the contact refrigerant 4 of the ice capsule forming tank 3 has been described.
  • the apparatus and method for cooling the cryopreservation containers 10 and 20 are not limited to this.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Thermal Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nutrition Science (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Sustainable Development (AREA)
  • Genetics & Genomics (AREA)
  • Mechanical Engineering (AREA)
  • Dentistry (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Freezing, Cooling And Drying Of Foods (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention porte sur un récipient de stockage à basse température (10), lequel récipient est hermétiquement scellé, après qu'un sac central étirable (12) qui fait exister un sujet à stocker (A) à l'intérieur de celui-ci et qui est rempli par un liquide (C) ayant un taux de compression supérieur à celui d'un liquide (D) et ayant une température de point de congélation inférieure à celle du liquide (D) est disposé dans un récipient externe (11), et qu'un espace entre le récipient externe (11) et le sac central (12) est rempli par le liquide (D), dont le volume est accru lors de la congélation. Ensuite, le récipient de stockage à basse température (10) est refroidi, et le sujet à stocker (A) est stocké à la température de stockage, qui est la température entre les points de congélation du liquide (C) et du liquide (D), et qui est inférieure à la température de point de congélation à la pression atmosphérique d'un liquide dans le sujet à stocker (A).
PCT/JP2012/051244 2011-01-20 2012-01-20 Procédé de stockage à basse température et récipient de stockage à basse température WO2012099249A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201280006028.5A CN103327839B (zh) 2011-01-20 2012-01-20 低温保存方法及低温保存容器
JP2012553787A JP5958913B2 (ja) 2011-01-20 2012-01-20 低温保存方法及び低温保存容器

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2011023296 2011-01-20
JP2011-023296 2011-01-20

Publications (1)

Publication Number Publication Date
WO2012099249A1 true WO2012099249A1 (fr) 2012-07-26

Family

ID=46515866

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2012/051244 WO2012099249A1 (fr) 2011-01-20 2012-01-20 Procédé de stockage à basse température et récipient de stockage à basse température

Country Status (3)

Country Link
JP (1) JP5958913B2 (fr)
CN (1) CN103327839B (fr)
WO (1) WO2012099249A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104206375A (zh) * 2013-05-29 2014-12-17 深圳华大方舟生物技术有限公司 封装胚胎的装置及方法

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106241011B (zh) * 2016-10-18 2018-07-06 宁波高智科技咨询服务有限公司 一种用于病患身体组织转移盛放装置
EP3634877A4 (fr) * 2017-05-08 2021-04-07 Claudia, Santamaria Dispositif de protection d'un récipient interne
CN111183095A (zh) * 2017-08-11 2020-05-19 加利福尼亚大学董事会 用于温度和压力控制的低温保存的方法和装置
CN111661448B (zh) * 2020-06-04 2022-04-15 淮北暴风工业设计有限公司 一种器官移植用运输箱

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06303954A (ja) * 1993-04-21 1994-11-01 Japan Steel Works Ltd:The 食品の低温保存方法
JP2010022269A (ja) * 2008-07-18 2010-02-04 Soka Univ 細胞の保存方法及びその装置
JP2010148401A (ja) * 2008-12-24 2010-07-08 Soka Univ 細胞保存容器

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5620732A (en) * 1995-06-07 1997-04-15 The Pillsbury Company Method of making ice cream
AU2001261666B2 (en) * 2000-05-18 2007-03-22 Supachill Technologies, Inc. Cooling method for controlled high speed chilling or freezing
US20050112252A1 (en) * 2003-11-20 2005-05-26 Gaurav Tewari Method to extend the shelf-life of food products using hydrostatic high-pressure processing
CN1921775A (zh) * 2004-02-20 2007-02-28 液体空气乔治洛德方法利用和研究的具有监督和管理委员会的有限公司 在交变大气压下处理食物的新方法
JP2006303954A (ja) * 2005-04-21 2006-11-02 Matsushita Electric Ind Co Ltd 撮像ユニットの製造装置及び方法
CN201403464Y (zh) * 2009-05-12 2010-02-17 上海善如水保鲜科技有限公司 一种果蔬不冻结减压保鲜装置

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06303954A (ja) * 1993-04-21 1994-11-01 Japan Steel Works Ltd:The 食品の低温保存方法
JP2010022269A (ja) * 2008-07-18 2010-02-04 Soka Univ 細胞の保存方法及びその装置
JP2010148401A (ja) * 2008-12-24 2010-07-08 Soka Univ 細胞保存容器

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104206375A (zh) * 2013-05-29 2014-12-17 深圳华大方舟生物技术有限公司 封装胚胎的装置及方法
CN104206375B (zh) * 2013-05-29 2016-04-20 深圳华大方舟生物技术有限公司 封装胚胎的装置及方法

Also Published As

Publication number Publication date
CN103327839B (zh) 2015-05-20
CN103327839A (zh) 2013-09-25
JP5958913B2 (ja) 2016-08-02
JPWO2012099249A1 (ja) 2014-06-30

Similar Documents

Publication Publication Date Title
JP5958913B2 (ja) 低温保存方法及び低温保存容器
Petzold et al. Ice morphology: fundamentals and technological applications in foods
CN112105863B (zh) 一种充装干式杜瓦罐的方法及装置
WO2009049448A1 (fr) Procédé permettant d'améliorer la qualité de décongélation de fruits et légumes surgelés au moyen d'ondes ultrasonores basse fréquence
JP5867893B2 (ja) 凍結方法
CN104542924A (zh) 一种果蔬保鲜的方法
EP4090157A1 (fr) Dispositifs et procédés pour une surfusion à haute stabilité de milieux aqueux et de matière biologique
KR101687448B1 (ko) 순간 침지 동결액 및 그 제조 방법
Fuchigami et al. Texture and structure of high-pressure-frozen gellan gum gel
CN202514486U (zh) 一种适用于液氮罐存贮细胞的容器
JP2007215535A (ja) 海水による予冷を導入する魚体用急速冷凍法
CN113200223A (zh) 远洋捕捞水产品保鲜设备
JP7217878B2 (ja) 保冷具、貨物、輸送機器、輸送方法及び保冷方法
JP2015223164A (ja) 凍結方法、冷却用ユニットおよび冷却システム
JP2022090501A (ja) 凍結ボックス
JP2016042808A (ja) 主としてカット野菜用の冷却容器
JPS6360989B2 (fr)
JP5939537B2 (ja) 魚卵の凍結保存法
JP2013100965A (ja) 食品の冷却・冷凍保存方法及び冷却・冷凍装置
JP2023011157A (ja) 液体冷凍保存用容器及び飲料の長期保存方法
Fuchigami Structural and Textural Quality of Pressure-shift-frozen Egg Custard Gel as Affected by Glucose, Trehalose or Sucrose
CN102927787B (zh) 速冻血浆的装置和方法
JPH06303954A (ja) 食品の低温保存方法
JPH034739A (ja) 生鮮食料品の貯蔵装置
CN102599144A (zh) 一种适用于液氮罐存贮细胞的容器

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 12737059

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2012553787

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 12737059

Country of ref document: EP

Kind code of ref document: A1