WO2006101237A1 - Freeze storage container and process for producing the same - Google Patents

Abstract

A freeze storage container is desired which has such freeze resistance that it does not break even at -196°C, which is the temperature of liquid nitrogen, can be molded in a relatively small wall thickness, and has excellent heat sealability. A freeze storage container comprising an adhesive fluoropolymer film is provided. This freeze storage container enables blood, scare cells, or a biotissue to be stored in an extremely-low-temperature environment, without breaking. The container can prevent liquid nitrogen from coming thereinto. It can prevent contamination by the bacteria, viruses, or the like present in the liquid nitrogen. Furthermore, it is free from the trouble that a freeze storage container breaks upon thawing due to the expansion of the liquid nitrogen which has come into the container.

Description

 Specification

 Cryopreservation container and manufacturing method thereof

 Technical field

 [0001] The present invention relates to a cryopreservation container and a method for producing the same. Specifically, a cryopreservation container comprising at least an adhesive fluorine-containing polymer film, preferably a laminate film including at least an adhesive fluorine-containing polymer film and a cold-resistant resin film, the laminate film. The invention relates to a cryopreservation container in which at least one surface of the film is an adhesive fluorine-containing polymer film.

 Background art

 [0002] In general, blood, rare cells, living tissues, and the like are stored at a cryogenic temperature of about 80 to 196 ° C. In particular, rare cells such as bone marrow cells and hematopoietic stem cells are effective in treating intractable diseases such as leukemia, and long-term storage techniques are required. For such storage at extremely low temperatures, it is general to place the container in a container containing the blood, rare cells, living tissue, etc., seal it, and store it mainly in liquid nitrogen.

 [0003] As cryopreservation containers used for storage, for example, polypropylene-made needles are commercially available at the laboratory level, and are inexpensive and easy to handle.

 In recent years, with the development of the cord blood bank, a bag-like container excellent in cold resistance and flexibility has been proposed. For example, a film having a laminated film strength of a polyimide film and a fluorinated ethylene propylene polymer film (Patent Document 1), a film having a film power of a copolymer of tetrafluoroethylene and ethylene (Patent Document 2), etc. have been proposed. ing. Furthermore, Patent Document 3 discloses a cryopreservation container formed of a film of an ethylene vinyl acetate copolymer that has been biaxially stretched by electron beam irradiation. Patent Document 4 discloses a cryopreservation container formed of a biaxially stretched crosslinked polyethylene film.

[0004] However, the fluorocarbon resins disclosed in Patent Documents 1 and 2 are required to be manufactured by heat sealing at a high temperature with a high melting point. Despite being manufactured under the conditions, the seal strength is weak, so it often breaks during cryopreservation. In addition, the stretched film disclosed in Patent Documents 3 and 4 is heat sealed. The film shrinks and wrinkles in the seal area and its surroundings due to the operation, and in actual use, it may often be damaged by rough handling. In addition, patent literature

Since the laminated film of 3 is bonded with an adhesive such as polyester, polyurethane and epoxy, the adhesive layer may harden during low-temperature storage, and the laminated film may peel off, making it suitable as a cryopreservation container. It is not a thing.

[0005] On the other hand, ultrahigh molecular weight polyethylene is very excellent in that it is impact resistant, abrasion resistant, self-swelling, chemical resistant, cold resistant and non-toxic, but it is extremely high !, melt viscosity Therefore, the film production method is usually made by cutting a powdery raw material resin into a block-shaped product by compression molding or the like. In the film manufactured as described above, since the surface state is rough, the molding force is not easy. In addition, there is a possibility that polyethylene on the surface of the film may fall off if there is a paste, and if used as a medical bag, it may cause contamination. In compression molding, air is taken in at the time of molding, and pinholes may occur in the film cut in the next process.

 In addition, when heat-sealing between sheets or between a sheet and a port part, the molecular weight of ultra-high molecular weight polyethylene is as high as 1 million or more, so the fluidity of polyethylene molecular chains is poor. Therefore, there is a problem in stable production of containers that are difficult to heat seal.

[0006] The cryopreservation container using the aromatic polyimide, the polyimide is an outer layer, the laminate of the non-oriented poly ethylene and the inner layer has been proposed (e.g., see Patent Document 5.) ₀ The product Sotai However, due to the heat fusion of the inner layer, the bag shape may not be possible, but due to the non-melting calorific properties of polyimide, lamination with the inner layer cannot be performed by thermal lamination using a heat roll or the like, and an adhesive is not used. Need to intervene and the above adhesive problem still remains

[0007] Patent Document 1: Japanese Patent Publication No.49-008079

 Patent Document 2: Japanese Utility Model Publication No. 55-055069

 Patent Document 3: Japanese Patent Publication No. 55-044977

 Patent Document 4: Japanese Patent Publication No. 62-057351

Patent Document 5: Japanese Patent Laid-Open No. 7-246230 Disclosure of the invention

 Problems to be solved by the invention

 [0008] From the above, there is a demand for the stable production of containers, particularly the development of cryopreservation containers excellent in low-temperature heat sealability. Furthermore, it has excellent sealing strength, and it is difficult to break the container during freezing storage, so the development of a cryopreservation container is required.

 Means for solving the problem

[0009] The present inventors propose to use an adhesive fluorine-containing polymer film for a cryopreservation container.

 In other words, the present invention

 [1] A cryopreservation container containing at least an adhesive fluorine-containing polymer film,

 [2] Adhesive fluorine-containing polymer film is composed of a fluorine-containing polymer having an adhesive site

The cryopreservation container according to [1],

 [3] Adhesive sites are carbon-carbon double bond, carbonyl group [—C (= 〇)], group or bond having a carbonyl group, hydroxyl group, cyano group, sulfonic acid group, and epoxy group The cryopreservation container according to [2], which is at least one selected from the group consisting of:

 [4] The adhesive fluorine-containing polymer has a reactive functional group as an adhesive site, and is a copolymer obtained by copolymerizing the following (A) and (B): [3] Described cryopreservation container:

(A) Fluorine-containing monomer having no reactive functional group

 (B) a fluorine-containing monomer having at least one reactive functional group,

 [5] The cryopreservation container according to [4], wherein the fluorine-containing monomer having no reactive functional group is represented by the following formula (1):

[Chemical 1] cx ^! ₂ = cx ²

(In the formula, X ¹ and X ² are each a hydrogen atom or a halogen atom, and Y is a hydrogen atom, a fluorine atom, a fluorine-containing alkyl group having 1 to 5 carbon atoms or a fluorine-containing alkyl group having 1 to 5 carbon atoms. Is a alkyl group),

 [6] Fluorine-containing monomer power without reactive functional groups Tetrafluoroethylene, Futsui vinylidene, 1,2-difunoleo-ethylene, hexafnoreo-propylene, perfluoronole (bulumetyl ether) and perfluoro ( The cryopreservation container according to [5], which is at least one selected from the group consisting of

 [7] A fluorine-containing monomer having a reactive functional group is represented by the following formula (2) [4] to [

[6] Cryopreservation container:

 [Chemical 2]

CX ¹ ;

 (¾

(In the formula, X ¹ and X ² are each a hydrogen atom or a halogen atom, Z is a hydroxyl group, a carboxyl group, a cyano group, a sulfonic acid group or an epoxy group, and R is a carbon number f.

 A fluorinated alkylene group having 1 to 40 carbon atoms, a fluorinated oxyalkylene group having 1 to 40 carbon atoms, or a fluorinated alkylene group having at least one ether bond having 1 to 40 carbon atoms).

 [8] The cryopreservation container according to [4], wherein the adhesive fluorine-containing polymer is represented by the following formula (3):

 [Chemical 3]

(In the formula, χ ¹ and ΧΊ are each a hydrogen atom or a halogen atom, and Υ ¹ and γ ² are a hydrogen atom, a fluorine atom, a fluorine-containing alkyl group having 1 to 5 carbon atoms, or a fluorine-containing group having 1 to 5 carbon atoms, respectively. Is an alkoxy group, and Ζ is a hydroxyl group, a carboxyl group, a cyano group, A sulfonic acid group or an epoxy group, and R is a fluorine-containing alkylene group having 1 to 40 carbon atoms,

 f

A fluorine-containing oxyalkylene group having 1 to 40 carbon atoms or a fluorine-containing alkylene group having at least one ether bond having 1 to 40 carbon atoms, and (1 + m) Zn force is 2000

[9] The adhesive fluorine-containing polymer has a fluorine-containing monomer unit derived from a fluorine-containing monomer and a fluorine-free monomer unit derived from a fluorine-free monomer. Container,

 [10] The cryopreservation container according to [9], wherein the fluorine-containing monomer is tetrafluoroethylene, and the fluorine-free monomer is ethylene.

 [11] It is composed of a laminated film including at least an adhesive fluorine-containing polymer film and a film other than the adhesive fluorine-containing polymer film, and the adhesive fluorine-containing polymer film is present on at least one outermost surface. The cryopreservation container according to [1],

[12] The cryopreservation container according to [11], wherein the film other than the adhesive fluorine-containing polymer film is a cold-resistant resin film,

 [13] The cold-resistant resin is at least one selected from the group consisting of ultrahigh molecular weight polyethylene, polyimide, polytetrafluoroethylene, ethylene-tetrafluoroethylene copolymer, and ethylene acetate butyl copolymer The cryopreservation container according to [12], [14] The cryopreservation container according to [13], wherein the cold-resistant rosin is polyimide,

[15] A method for producing a cryopreservation container, wherein at least an adhesive fluorine-containing polymer film is formed into a bag shape by heat sealing,

And [16] A laminated film including at least an adhesive fluorine-containing polymer film and a film other than the adhesive fluorine-containing polymer film, wherein the adhesive fluorine-containing polymer film is present on at least one outermost surface is heated. The present invention relates to the method for producing a cryopreservation container according to [15], which is formed into a bag shape with a seal.

The invention's effect

Since the cryopreservation container of the present invention can be heat-sealed even at a low temperature, the container can be stably manufactured without manufacturing the noggles under severe conditions. In addition, because the heat seal strength is strong, it is possible to prevent the container from being damaged during cryopreservation. Precious biological samples can be stably stored.

 BEST MODE FOR CARRYING OUT THE INVENTION

 [0011] The cryopreservation container of the present invention includes at least an adhesive fluorine-containing polymer film.

 The adhesive fluorine-containing polymer film is a film formed of a polymer having at least one fluorine atom in the main chain and Z or side chain, and the film adheres to a substrate made of an organic material. It has a function. Here, the term “adhesion” means that the above-mentioned adhesive fluorine-containing polymer film is bonded to the organic material by physical and Z or chemical bonding. From the viewpoint of bonding strength, chemical bonding is preferable. It is not limited to this. Examples of the chemical bond include a covalent bond, an ionic bond, a coordination bond, a hydrogen bond, and an intermolecular force. From the viewpoint of bond strength, the bond is preferably a covalent bond and an ionic bond, and more preferably a covalent bond. Is not limited to this force

[0012] The organic material includes polyethylene, polycarbonate, polystyrene, and polyvinyl chloride.

-Films made of cold-resistant resin such as rubber, polyacetate butyl, polyester and ultra high molecular weight polyethylene, polyimide, polytetrafluoroethylene, ethylene-tetrafluoroethylene copolymer and ethylene vinyl acetate copolymer, General purpose resin molding products such as tubes, synthetic fibers, synthetic rubbers and solid materials, and natural organic materials such as natural rubber, natural fibers, wood, papers and leathers. Includes the molecule itself. Among them, from the viewpoint of improving the cold resistance of the cryopreservation container itself, it is preferable that it can be adhered to an organic material having a function of adhering to the adhesive fluorine-containing polymer film itself and the cold resistant resin film. It is not limited to this.

 [0013] From the viewpoint of molding processability, the molecular weight of the above-mentioned adhesive fluorine-containing polymer is about 1,000 to 1,000,000, preferably <is about 2,000 to 500,000 in terms of number average molecular weight. More preferred, <is a force that is about 5,000-300,000, but is not limited to this.

[0014] The above-mentioned adhesive fluorine-containing polymer film can be produced by a person skilled in the art selecting a suitable production method in a timely manner according to the molecular structure of the adhesive fluorine-containing polymer, the glass transition temperature, the melting point, and the like. For example, compression molding, injection molding, extrusion molding, τ die molding, inflation Examples include sill molding and solvent casting, and compression molding is preferred from the viewpoint of molding processability, but is not limited thereto. The film thickness of the adhesive fluorine-containing polymer film is about 10 to: LOO / zm, preferably 10 to 50 μm, and particularly preferably 10 to 30 μm, from the viewpoint of thermal conductivity to the contents. However, the present invention is not limited to this.

 [0015] Preferred examples of the above-mentioned adhesive fluorine-containing polymer include a copolymer containing the following (A) and (B).

 (A) A fluorine-containing monomer having no reactive functional group.

 (B) A fluorine-containing monomer having at least one reactive functional group.

 [0016] The reactive functional group in the present invention is a functional group capable of adhering to the base material made of the organic material through a covalent bond, an ionic bond, a coordinate bond, a hydrogen bond, and the like. Examples thereof include a carboxyl group, a cyano group, a sulfonic acid group, and an epoxy group. Among them, a hydroxyl group that is easily activated by heat is preferable, but is not limited thereto.

 [0017] The fluorine-containing monomer is a monomer in which at least one hydrogen atom in the main chain and Z or side chain of the copolymer obtained by polymerization is substituted with fluorine. Examples thereof include fluorine ethylenic monomers, fluorine-containing ester monomers, and fluorine-containing wholly aromatic monomers. Among these, a fluorine-containing ethylene monomer is preferable from the viewpoint of easy availability and molding moldability of the resulting copolymer, but is not limited thereto.

 [0018] The copolymer may be a two-component system obtained by polymerizing at least one of each of (A) and (B). For example, a two-component system consisting of one (A) monomer and one (B) monomer, a three-component system consisting of two (A) monomers and one (B) monomer, and Examples include a three-component system composed of one type of monomer (A) and two types of monomer (B). Of these, a two- or three-component system is preferable from the viewpoint of production cost, but is not limited thereto.

[0019] Examples of the copolymer include radical copolymerization, anion copolymerization, cationic copolymerization, emulsion copolymerization, and plasma copolymerization. Depending on the monomer structure, polarity, type of solvent, etc. Those skilled in the art can select it in a timely manner. Of these, radical copolymerization is preferred from the viewpoint of ease of production! /, But is not limited to this! /.

 [0020] Examples of the form of the copolymer include a random copolymer, a block copolymer, a graft copolymer, and an alternating copolymer. Among these, random copolymer is preferred from the viewpoint of manufacturability, but it is not limited to this.

 [0021] Further, the abundance ratio (copolymerization ratio) between (A) and (B) in the above copolymer is (1) from the viewpoint of the formation caloric property of the copolymer. A) is 1 to 2000, preferably 100 to 2000, but is not limited thereto.

 [0022] The (A) fluorine-containing monomer having no reactive functional group is a main chain and a Z or side chain of a copolymer that does not have the above-mentioned reactive functional group and is obtained by polymerization. A copolymer in which at least one hydrogen atom is substituted with fluorine is obtained. Among these, a fluorine-containing ethylenic monomer having no reactive functional group is preferred from the viewpoint of easy availability and molding processability of the resulting copolymer, but is not limited thereto.

 [0023] The (A) fluorine-containing ethylenic monomer having no reactive functional group contains at least one fluorine atom, and examples thereof include a monomer represented by the following formula (1).

[0024] [Chemical 4]

し 入 2— し 入 Π 、

 Y

(Wherein X ¹ and X ² are each a hydrogen atom or a halogen atom, Y is a hydrogen atom, a fluorine atom, a fluorinated alkyl group having 1 to 5 carbon atoms or a fluorinated oxyalkyl group having 1 to 5 carbon atoms. )

[0025] Monomers represented by the above formula (1) include, for example, tetrafluoroethylene, vinylidene fluoride, 1,2-difunoleochloroethylene, hexafluororeopropylene, perfnoreo (bulurmethyl ether). ) And perfluoro (bulupropyl ether), preferably tetrafluoroethylene, vinylidene fluoride, 1,2-difluorochloroethylene and perfluoro (bulupropyl ether). Above all, from the viewpoint of easy availability and molding cacheability of the resulting copolymer, tetrafluorobenzene. Tylene, vinylidene fluoride, hexafluoropropylene and perfluoro (bulupropyl ether) are preferred, but not limited thereto.

 [0026] On the other hand, (B) the fluorine-containing monomer having at least one kind of reactive functional group has at least one reactive functional group described above, and the main chain of the copolymer obtained by polymerization and Z or A copolymer in which at least one hydrogen atom in the side chain is substituted with fluorine is obtained. Among these, a fluorine-containing ethylenic monomer having a reactive functional group is preferred from the viewpoint of easy availability and molding processability of the resulting copolymer, but is not limited thereto.

 [0027] (B) Fluorine-containing ethylenic monomer having at least one reactive functional group

And a monomer represented by the formula (2).

[0028] [Chemical 5]

(In the formula, X ¹ and X ² are each a hydrogen atom or a halogen atom, Z is a hydroxyl group, a carboxyl group, a cyano group, a sulfonic acid group or an epoxy group, and R is a carbon number f.

A fluorinated alkylene group having 1 to 40 carbon atoms, a fluorinated oxyalkylene group having 1 to 40 carbon atoms, or a fluorinated alkylene group having at least one ether bond having 1 to 40 carbon atoms.)

[0029] Examples of the monomer represented by the above formula (2) include perfluoro- (4 oxa 5 monohexenol) (formula (4)), perfluoro (1, 1-dihydro-6 heptenol) (formula (5)), Perfluoro (1, 1, 9, 9—Tetrahydride 2,5 Bistrifluoromethyl-3, 6-Dioxer 8-Nenol) (Formula (6)), Perfluoro (4-Oxa-5-hexenoic acid) (Formula (7)), perfluoro- (3,6-dioxa-4-trifluoromethyl-7-otatheno-tolyl) (formula (8)), perfluoro- (1,1-dihydride 3 Ntenosulfonic acid) (formula (9)) and 1,2 epoxy-perfluoro- (1,1,2 trihydr draw 6 pentene) (formula (10)), and the like. Perfluoro- (4-oxa 5-hexenol) (formula (4)), perfluoro (1,1-dihydro-6-heptenol) (formula (5)) and perfluoro- (1, 1, 9, 9-tetrahydride) Mouth 2,5 Bistrifluoromethyl 1,3,6 Dioxa 1 8None) (formula (7)) and other forces having a hydroxyl group The present invention is not limited to this.

[0030] [Chemical 6]

(5) [0031] [Chemical 7]

(Five)

[0032] [Chemical 8]

[0033] [Chemical 9]

CF ₂ = CF »OCF2CF ₂ CF ₂ » COOH (7)

[0034] [Chemical 10]

CF ₃ ^(¾

[0035] [Chemical 11]

CH ₂ = CF-OCF H ₂ -S0 ₄ H (9)

[0036] [Chemical 12] 丄 n 2 丄 2 HA (10)

 o

[0037] A preferred form of the adhesive fluorine-containing polymer in the present invention is exemplified by a copolymer represented by the following formula (3) from the viewpoint of ease of production, but is not limited thereto. It is not something.

 [0038] [Chemical 13]

(In the formula, X ¹ and X ² are each a hydrogen atom or a halogen atom, and γ ¹ and γ ² are each a hydrogen atom, a fluorine atom, a fluorine-containing alkyl group having 1 to 5 carbon atoms, or a carbon atom having 1 to 5 carbon atoms. A fluorine alkoxy group, z is a hydroxyl group, carboxyl group, cyano group, sulfonic acid group or epoxy group, R is a fluorine-containing alkylene group having 1 to 40 carbon atoms, f

 A fluorine-containing oxyalkylene group having 1 to 40 carbon atoms or a fluorine-containing alkylene group having at least one ether bond having 1 to 40 carbon atoms, and (1 + m) Zn force to 2000) above (l + m ) If Zn exceeds 2000, sufficient adhesion may not be obtained.

 [0039] As a more preferable form of the adhesive fluorine-containing polymer in the present invention, a force represented by a copolymer represented by the following formula (11) from the viewpoint of ease of production and heat seal strength: It is not limited to.

 [0040] [Chem. 14]

(Where x ¹ and x ² are hydrogen atoms or halogen atoms, respectively, and γ ¹ and γ ² are Respectively a hydrogen atom, a fluorine atom, a fluorine-containing alkyl group having 1 to 5 carbon atoms or 1 carbon atom

~ 5 fluorine-containing alkoxy group, R is carbon number 1

 f is a fluorine-containing alkylene group having 40 to 40 carbon atoms, a fluorine-containing alkylene group having 1 to 40 carbon atoms, or a fluorine-containing alkylene group having at least one ether bond having 1 to 40 carbon atoms, and (1 + m) Zn [0041] As a particularly preferred form of the adhesive fluorine-containing polymer in the present invention, polytetrafluoropolymer is preferable from the viewpoint of easy availability of monomers, ease of production, and heat seal strength. Co-polymerization of ethylene and Z or perfluoro (bulpropyl ether) with perfluoro- (1, 1, 9, 9—tetrahydric 2,5—bistrifanololeolomethinole 3, 6—dioxa 8—nonenol) Combined force It is not limited to this. In addition, the copolymer composition ratio of perfluoro (1,1,9,9-tetrahydride mouth-2,5-bistrifluoromethyl-3,6-dioxa 1-8- Nonenol) monomer unit

1, polytetrafluoroethylene monomer unit and Z or perfluoro

(Bulupropyl ether) A force with the total of monomer units of about 2 to 2000, preferably about 4 to 2000, but is not limited thereto.

[0042] In the present invention, it is preferable that the adhesive fluorine-containing polymer film has an adhesive fluorine-containing polymer force having an adhesive site.

 The above-mentioned adhesive fluorine-containing polymer should have the above-mentioned reactive functional group as an adhesive site.

 The above-mentioned adhesive fluorine-containing polymer film may form an adhesive fluorine resin layer described later. The above-mentioned adhesive fluorine-containing polymer may constitute an adhesive fluorine resin.

 [0043] The adhesive fluorine resin layer in the present invention is made of an adhesive fluorine resin.

 In the present specification, the adhesive fluorine resin is preferably a fluoropolymer having an adhesive site.

In the present specification, the fluoropolymer is a polymer having a fluorine-containing monomer unit derived from a fluorine-containing monomer in the main chain. Further, the fluoropolymer may or may not have a fluorine-free monomer unit derived from a fluorine-free monomer. In the present specification, the “monomer unit” for the fluoropolymer means a part of a polymer molecular structure derived from a monomer. For example, a tetrafluoroethylene unit is represented by one (CF 3 -CF 3) —.

 twenty two

 [0045] The fluorine-containing monomer is not particularly limited as long as it is a polymerizable compound having a fluorine atom, and examples thereof include tetrafluoroethylene [TFE], vinylidene fluoride [VdF], black trifluoroethylene [ CTFE], vinyl fluoride [VF], hexafluoropropylene [HFP], hexafluoroisobutene, perfluoro (alkyl butyl ether) [PAVE] s, the following general formula (i):

CH = CX ³ (CF) X ⁴ (i)

 2 2 n

(Wherein x ³ represents a hydrogen atom or a fluorine atom, X ⁴ represents a hydrogen atom, a fluorine atom or a chlorine atom, and n represents an integer of 1 to 10), etc. Is mentioned.

 [0046] The fluorine-free monomer is not particularly limited as long as it is a compound copolymerizable with the fluorine-containing monomer and does not have a fluorine atom. For example, ethylene [Et], propylene, 1-butene, 2-butene, Examples include vinyl chloride and vinylidene chloride.

 [0047] Examples of the fluoropolymer include the following copolymer (I) and the following copolymer (Π).

 (I) a copolymer obtained by polymerizing at least TFE and Et,

 (Ii) At least TFE and the following general formula (ii)

CF = CF-R ² (ii)

 2 f

(Wherein R ² represents CF or —OR ¹ , and R ¹ represents a perfluoroalkyl having 3 to 5 carbon atoms, f 3 ff

 Represents a ru group. A copolymer obtained by polymerizing at least one monomer represented by

[0048] Examples of the copolymer (I) include at least a copolymer having a TFE unit of 20 to 80 mol% and an Et unit of 80 to 20 mol%.

In the present specification, the mol% for each monomer unit is the adhesive site-containing monomer unit described later, out of the total number of moles of the monomer from which the monomer unit constituting the molecular chain of the copolymer is derived. the model number Le excluding moles of monomer was decided that derived as 100 mole _^0/0, a ratio of moles of monomer each monomer unit was to derived occupied in the 100 mole _^0/0 is there. The mol% for each monomer unit is a value obtained from an F-NMR chart.

[0049] The copolymer (I) may have other monomer units derived from other copolymerizable monomers in addition to the TFE unit and Et unit in the main chain. As the monomer, a monomer of a type corresponding to the use of the obtained laminated film can be appropriately selected and used for copolymerization.

Examples of other monomers include HFP, CTFE, propylene, and the following general formula (iii): CX ⁵ = CX ⁶ (CF) X ⁷ (iii)

 2 2 n

(In the formula, X ⁵ and X ⁶ are the same or different and each represents a hydrogen atom or a fluorine atom; X ⁷ represents a hydrogen atom, a fluorine atom or a chlorine atom; and n represents an integer of 1 to 10. ) Monomers represented by the following general formula (iv):

CF = CF— OR ¹ (iv)

 2 f

(Wherein R ¹ represents a perfluoroalkyl group having 1 to 5 carbon atoms) f

 In general, one or more of these are used.

These and other monomer units may be filed those having a ratio of 0-20 mole _^0/0 of the monomeric units 100 mole _^0/0 constituting the molecular chain of the copolymer (I).

 [0050] As the above fluoropolymer, copolymer (I) is preferred in terms of heat resistance, chemical resistance, weather resistance, electrical insulation, low liquid chemical permeability, non-adhesiveness, etc. EtZTFEZHFP copolymer is more preferable because of its excellent properties, chemical resistance, weather resistance, electrical insulation, low chemical permeability, non-adhesiveness, low-temperature processability, transparency and the like. The HFP unit in the above EtZTFEZHFP copolymer preferably has a lower limit of 8 mol%, more preferably 5 to 20 mol%, and a more preferable upper limit of 17 mol%. In addition to the monomer units derived from Et, TFE, and HFP, the EtZTFEZHFP copolymer is one kind of the above-mentioned other monomers other than the HFP unit, as long as the EtZTFEZHFP copolymer is preferred and does not lose its properties. Or you may have two or more.

 In the present specification, the “adhesive site” means a functional group having affinity or reactivity with the above-mentioned organic material such as polyimide [PI] film.

In this specification, “affinity” means a chemical structure such as hydrogen bond, van der Waals force, etc. It means a property that shows an interaction with an organic material such as a PI film that does not lead to a change, and “reactivity” means a property that changes a chemical structure such as a functional group.

 [0052] The adhesive site is usually one that the fluoropolymer has in the main chain or side chain.

 The adhesive site is not particularly limited, and examples thereof include a carbon-carbon double bond, a force sulfonyl group [-C (= O)], a group having a carbo group or a bond, and the like. In the fluoropolymer having a site, there may be only one type of adhesive site, or two or more types of adhesive sites! /.

 The “adhesive site” may be the reactive functional group described above.

[0053] Examples of the group or bond having a carbonyl group include a carbonate group, a halogenoformyl group, a formyl group, a carboxyl group, a carbo-loxy group [C (= o) o], an acid anhydride group [- C (= 0) 0— C (= 0) —], isocyanate group, amide group [— C (= 0) —NH], imide group [— C (= 0) — NH— C (= 0) —] , Urethane bond [― NH— C (= 0) 0], force rumomoyl group [NH—C (= 0) —], force rumomoyloxy group [NH—C (= 0)

 twenty two

 O], ureido group [NH—C (= 0) —NH], oxamoyl group [NH—C (= 0) —C

 twenty two

 (= o) —] and the like.

 The group or bond having a carbonyl group is preferably a carbonate group, a halogenoformyl group or the like from the viewpoint of easy introduction and high reactivity.

 [0054] The carbonate group is a group having a bond represented by [-OC (= 0) 0],-OC (= 0) 0- R group (wherein R is an organic group, group IA Represents an atom, a group IV atom, or a group VIIB atom.) Examples of the organic group for R in the above formula include an alkyl group having 1 to 20 carbon atoms, an alkyl group having 2 to 20 carbon atoms having an oxygen molecule constituting an ether bond, and preferably 1 to 8 carbon atoms. Or an alkyl group having 2 to 4 carbon atoms and having an oxygen molecule constituting an ether bond. Examples of the carbonate group include one OC (= 0) 0—CH, one OC (= 0) 0—C H, one OC (= 0) 0

 3 3 7

 —CH, —OC (= 0) 0—CH CH OCH CH and the like.

 8 17 2 2 2 3

[0055] The halogenoformyl group is represented by COY (wherein Y represents a VIIB group atom), and -COF, -COC1, and the like are preferable. [0056] The number of the adhesive sites may be appropriately selected depending on the type, shape, application, required adhesive strength, difference in the type of the fluoropolymer, and the like. 10 per ⁶ 3 ~: LOOO. When counting the number of carbonyl groups, the number of the above-mentioned adhesive sites is usually 150 or more, preferably 250 or more, more preferably 300 or more per 1 × 10 ⁶ main chain carbon atoms. .

 In the present specification, the number of the “adhesive sites” is measured by performing an infrared absorption spectrum analysis in accordance with the method for measuring the number of carbonyl group-containing functional groups described in International Publication No. 99Z45044. is there.

[0057] Examples of the adhesive fluorine resin include a fluorine-containing ethylenic polymer having a carbonyl group-containing functional group described in International Publication No. 99Z45044 pamphlet.

 The adhesive fluorine resin can be usually obtained by introducing an adhesive site in the production of a fluoropolymer by polymerization, but the method for introducing the adhesive site is not particularly limited. For example, (1) A method of copolymerizing a monomer containing an adhesive moiety, (2) polymerization in an aqueous medium such as emulsion polymerization in the presence of a polymerization initiator having an adhesive moiety, and the polymerization initiator at the end of the polymer chain (3) A method in which the carbon-carbon single bond in the polymer chain is changed to a double bond during polymerization or by heating after polymerization, etc. Is mentioned.

[0058] In the method of (1), for example, an adhesive site-containing monomer, a fluorine-containing monomer of a type and a composition depending on the target adhesive fluorine resin, and optionally a fluorine-free monomer are known. Can be carried out by copolymerization.

 The copolymerization method is not particularly limited, and is, for example, random copolymerization in which an adhesive site-containing monomer is introduced into the system when a polymer chain is formed with another comonomer such as a fluorine-containing monomer. Alternatively, block copolymerization or graft copolymerization may be used. Examples of the draft copolymerization include a method in which an unsaturated carboxylic acid described later is attached to a fluoropolymer.

[0059] The "adhesive moiety-containing monomer" means a polymerizable compound having an adhesive moiety, and may or may not have a fluorine atom. In this specification, The above-mentioned “fluorine-containing monomer” and “fluorine-free monomer” are those having the above-mentioned adhesive part.

 [0060] Examples of the adhesive part-containing monomer include, when the adhesive part is a group having a carbo group or a bond, perfluoroacrylic acid fluoride, 1 fluoroacrylic acid fluoride, acrylic acid fluoride, 1 Monomers having fluorine such as trifluoromethacrylic acid fluoride and monofluorobutenoic acid; monomers having no fluorine such as acrylic acid, methacrylic acid, acrylic acid chloride and beylene carbonate.

 [0061] Examples of the adhesive site-containing monomer further include unsaturated carboxylic acids.

 In the present specification, the above unsaturated carboxylic acids means at least a carbon-carbon unsaturated bond (hereinafter also referred to as “copolymerizable carbon-carbon unsaturated bond”) that enables copolymerization in at least one molecule. Even if it is a compound having one and having at least one carbo-loxy group [—C (= 0) —O] in one molecule, the above-mentioned copolymerizable carbon-carbon unsaturated bond Is preferred to have 1 per molecule! /.

 Examples of the unsaturated carboxylic acids include aliphatic unsaturated carboxylic acids and acid anhydrides thereof. The aliphatic unsaturated carboxylic acid may be an aliphatic unsaturated monocarboxylic acid, or may be an aliphatic unsaturated polycarboxylic acid having two or more carboxyl groups.

 As said aliphatic unsaturated monocarboxylic acid, C3-C20 aliphatic monocarboxylic acid etc., such as propionic acid, acrylic acid, methacrylic acid, crotonic acid, those acid anhydrides, etc. are mentioned, for example. Examples of the aliphatic unsaturated polycarboxylic acid include maleic acid, fumaric acid, mesaconic acid, citraconic acid [CAC], itaconic acid, aconitic acid, itaconic anhydride [IAH], and citraconic anhydride [CAH]. .

[0062] As the polymerization initiator in the above method (2), diisopropyl peroxy carbonate, di-n-propyl peroxydicarbonate, t-butyl peroxy isopropyl carbonate, bis (4-tert-butyl cyclohexane) are used. Hexyl) peroxydicarbonate, di-2-ethylhexyloxydicarbonate and the like.

[0063] The adhesive fluorine resin has a melting point of preferably 200 ° C or lower, more preferably 180 ° C or lower, from the viewpoint of the sealing property of the obtained laminated film. In the present specification, the melting point is a temperature at the maximum value of the melting peak obtained by measuring with a differential scanning calorimeter (Seiko Co., Ltd.) at a heating rate of 10 ° CZ.

[0064] The adhesive fluorine-containing polymer film in the present invention may be in the form of a laminated film with a film other than the adhesive fluorine-containing polymer film. The laminated film in the present invention is a laminate of at least an adhesive fluorine-containing polymer film and a film other than the adhesive fluorine-containing polymer film, and the adhesive fluorine-containing polymer film is at least one outermost surface. As long as it exists in The number of layers may be two or more, but from the viewpoint of thermal conductivity to the contents, the force is 2 to 5 layers, preferably 2 and 3 layers, but is not limited thereto.

 [0065] The film thickness of the laminated film depends on the number of layers. For example, when the film is a two-layer film of an adhesive fluorine-containing polymer film provided on one side of the film other than the adhesive fluorine-containing polymer film, From the viewpoint of thermal conductivity to the contents, the total film thickness is about 20 to 200 111, preferably about 20 to: LOO μm, particularly preferably about 20 to 60 μm, but is not limited thereto. It is not done.

 [0066] The thickness of the adhesive fluorine resin layer is preferably 5 to: LOO / zm, more preferably 10 µm or more and 50 µm or less.

[0067] When the laminated film of the present invention is produced by the above-mentioned thermal lamination method, the adhesion strength (X) is generally 200 NZm or more, preferably 300 NZm or more, more preferably 40.

It can be over ONZm.

 In the present specification, the adhesive strength (X) is obtained by cutting a laminated film into a width of 10 mm, and using an edge of an adhesive fluororesin layer at the end and a film other than an adhesive fluorine-containing polymer film such as a PI film. This is the strength required when peeled off to make a grip and peeled 180 ° at a speed of 25 mmZ on a Tensilon universal testing machine.

[0068] A film other than the above-mentioned adhesive fluorine-containing polymer film is not particularly limited unless it is a film having the above-mentioned adhesive fluorine-containing polymer power, but is preferably a cold-resistant resin. .

[0069] The cold-resistant rosin refers to a rosin excellent in impact resistance at about -40 degrees or less, preferably about -80 degrees or less. For example, ultra high molecular weight polyethylene, polyimide, polytetraflur Examples include fluoroethylene, ethylene-tetrafluoroethylene copolymer, and ethylene vinyl acetate copolymer. From the viewpoint of thermal conductivity to the contents and cold resistance, polyimide, polytetrafluoroethylene, and ethylene Polytetrafluoroethylene copolymer is more preferable than polyimide. In addition, the molecular weight of cold-resistant rosin has a number average molecular weight of about 1,000 to 1,000,000, preferably <2,000 to 500,000, more preferably < The force that is about 5,000-300,000 is not limited to these.

 The above impact resistance is evaluated using a free fall dart impact test method (staircase method (JIS Κ 7124-1)) for a resin film or sheet immediately after removal from a frozen environment. Is. As the above-mentioned resin having excellent impact resistance, the 50% fracture energy (Ε50) in the above-mentioned staircase method (liquid nitrogen temperature) is 0.1 or more, preferably 0.2 or more, more preferably 1.0. The above is preferable.

 [0070] The polyimide is not particularly limited as long as it has a heat-resistant polymer force having an imide bond in the main chain, for example, a non-thermoplastic polyimide having only an imide bond in the main chain, a wholly aromatic polyimide, an organic solvent soluble Forces including, but not limited to, polyimide, polyetherimide, and polyimideamide.

 [0071] In addition, the cold-resistant resin film includes, for example, a force obtained by forming a cold-resistant resin by a hot melting method, an extrusion method or a compression method and a solvent casting method at a high temperature and a high pressure. Is not to be done.

[0072] Examples of the lamination of the adhesive fluorine-containing polymer film and the film other than the adhesive fluorine-containing polymer film in the laminated film include a heat laminating method, a heat compression method, a high-frequency heating method, and a solvent casting method. . Of these, the thermal lamination method is preferable from the viewpoint of ease of production, but is not limited thereto. Since the adhesive fluorine-containing polymer film adheres firmly to a film other than the above-mentioned adhesive fluorine-containing polymer film, the adhesive fluorine-containing polymer film may be peeled off from the film other than the adhesive fluorine-containing polymer film. Absent. The temperature condition in the thermal laminating method is a force of about 200 to 300 ° C., preferably about 200 to 250 ° C., from the viewpoint of safety in production and the like, but is not limited thereto. [0073] The laminated film in the present invention can be prepared by laminating a film other than the adhesive fluorine-containing polymer film such as the above-mentioned PI film and an adhesive fluorine resin. Laminating of a film other than the adhesive fluorine-containing polymer film and the adhesive fluorine resin can be performed, for example, by an extrusion lamination method, or bonding with a film other than the adhesive fluorine-containing polymer film such as a PI film. It is also possible to carry out by adhering a functional fluorine resin by thermocompression bonding or the like.

 [0074] In the extrusion lamination, for example, an extrusion process (a) in which the above-mentioned adhesive fluorine resin is melted and extruded onto a film other than the above-mentioned adhesive fluorine-containing polymer film such as the above-mentioned PI film (a), PI A crimping step (b) of crimping a film other than an adhesive fluorine-containing polymer film, such as a film, and an extruded adhesive fluororesin sandwiched between rolls, and a scraping step of scraping the resulting laminate (c) In general, the extrusion process (a), the crimping process (b), and the scraping process (c) are performed in this order.

 [0075] The extrusion temperature in the extrusion step (a) depends on the type of film other than the adhesive fluorine-containing high molecular film such as the PI film used and the adhesive fluorine resin, the thickness of the target laminated film, etc. In general, it is preferably at least the melting point of the adhesive fluororesin used and less than the decomposition temperature in that a laminated film having a high interlayer adhesive strength can be obtained.

 [0076] In the extrusion step (a), the molten adhesive fluorine resin is extruded onto a film other than the adhesive fluorine-containing polymer film such as the PI film. A force that can be appropriately set depending on the composition, thickness, etc. of the film other than the above-mentioned adhesive fluorine-containing polymer film such as the PI film. For example, 0.1 to: LOOmZ it can.

 [0077] The extrusion lamination, particularly the extrusion step (a), is performed in an inert gas and includes an adhesive property such as a Z or PI film in that a laminated film having a high interlayer adhesive strength is obtained. It is preferable to remove water by drying or preheating a film other than the fluoropolymer film in advance.

In the present invention, the extrusion lamination is present in the adhesive fluorocarbon resin by the extrusion process. However, it is considered that the above-mentioned extrusion lamination is performed in an inert gas, and a film other than an adhesive fluorine-containing high molecular film such as a Z or PI film. It is considered that the above adhesiveness can be sufficiently exhibited when moisture is removed by drying or preheating the material in advance.

 [0078] The process conditions other than the above-described extrusion step (a) in the above-mentioned extrusion lamination are the types of films other than the adhesive fluorine-containing polymer film such as the PI film to be used, and the types of adhesive fluorine resin. Depending on the thickness of the laminated film, etc., it can be appropriately set according to a known method.

 [0079] When a film other than the above-mentioned adhesive fluorine-containing polymer film such as the above-mentioned PI film and the above-mentioned adhesive fluorine resin are laminated by thermocompression bonding, it is generally adhered by a known extrusion method or the like. After forming the fluorinated fluoro resin into a film, the obtained adhesive fluorinated resin film and a film other than an adhesive fluorinated polymer film such as a PI film are overlaid and heated and pressure-bonded. Can do.

 The thermocompression bonding is preferably performed at a temperature of 120 to 300 ° C. The temperature is more preferably V lower limit of 140 ° C and more preferably upper limit of 280 ° C.

 When the films other than the adhesive fluorine-containing polymer film such as the above-mentioned PI film are laminated by thermocompression bonding or the like, they may be preheated prior to lamination or pre-dried. Also, when bonding each layer by thermocompression bonding, etc., in order to improve interlayer adhesion, it is possible to heat and age after bonding!

 The heating for the aging is preferably performed at 200 to 280 ° C.

[0080] The laminated film is formed into a container or bag shape by a heat sealing method or the like. Specifically, two laminated films are stacked so that the adhesive fluorine-containing polymer films are in contact with each other, and heat sealing is performed. The inner volume of the container is generally about 2 to 200 ml, but is not limited to this. The width of the heat seal is not limited to this, which is about 2 to 20 mm, preferably about 5 to 15 mm, from the viewpoint of the sealing strength between the laminated films. Further, the heat seal temperature is about 180 to 250 ° C, preferably about 200 to 220 ° C, from the viewpoint of the sealing strength between the laminated films. General-purpose fluororesin films cannot be sealed at temperatures as low as 180-250 ° C. In other words, the present invention Since the adhesive fluorine-containing polymer film has good heat sealability at low temperatures, the cost in the production process is also low.

 [0081] The cryopreservation container of the present invention contains, for example, blood components such as red blood cells, platelets, and plasma, and biological samples such as bone marrow fluid, other body fluids, and cell suspensions, and the contents when the cryopreservation container is damaged. It may be further packaged to protect materials and prevent liquid nitrogen from entering the cryopreservation container. As these packages, packages made of perfluoroethylenepropene copolymer are generally used, but are not limited thereto. At this time, the air between the cryopreservation container and the package can be easily degassed by using, for example, an auxiliary tool disclosed in Japanese Patent Application Laid-Open No. 2000-185716. It is not limited.

 [0082] The cryopreservation container of the present invention can be produced by forming the laminated film into a container or bag shape by a heat seal method or the like. When the obtained cryopreservation container uses a laminated film in which an adhesive fluoropolymer film is present on at least one outermost surface as the above-mentioned laminated film, the cryopreservation container contains adhesiveness on at least one outermost surface. The container can be a container having a fluoropolymer film, and is preferably a container having an adhesive fluorine-containing polymer film at least on the outermost surface inside the container.

 [0083] The cryopreservation container of the present invention is capable of withstanding even at extremely low temperatures of -80 to 196 ° C. Forces Blood components such as erythrocytes, platelets and plasma, bone marrow fluid, other body fluids and cell suspensions In the case of actual storage, it is preferable to cool gradually in order not to damage these tissues. For example, the power is not limited to this, for example, a method of once cooling it to about −80 degrees with a deep freezer or the like and then storing it in liquid nitrogen. When using stored blood, cells, etc., the force that can be thawed by a heating means such as a 37-40 ° C. warm bath is not limited thereto.

[0084] When the biological sample is stored, a commercially available storage solution can be appropriately used. For example, when cells are stored, examples of the storage solution include DMEM medium, RPMY1640 medium, 199 medium, and phosphate buffer. Preferably, about 0.5 to 2% by volume of albumin may be added. More preferably, dimethyl sulfoxide (DMSO) is used as a freezing protection agent. A final concentration of about 5-20% by volume may be added. When organs are preserved, Eurocollins solution and UW solution can be used. Preferably, dimethyl sulfoxide (DMSO) may be added as a frost damage protective agent to a final concentration of about 5 to 20% by volume. The selection and preparation of these preservation solutions are not particularly limited because they can be appropriately selected by those skilled in the art.

 [0085] Examples of the method for freezing the cryopreservation container of the present invention containing or storing a biological sample include a method of freezing the cryopreservation container containing or storing a biological sample at 0 ° C or lower. As the freezing method, a method of freezing at 80 ° C. or lower is preferable. In the freezing method, it is preferable that the cryopreservation container is gradually cooled to a desired temperature and frozen in order not to damage the biological sample. Examples of such a freezing method include a method in which the cryopreservation container is once cooled to about 80 ° C. with a deep freezer (freezer) and then immersed in liquid nitrogen. The cryopreservation container may contain the above-described storage solution as necessary. The above cryopreservation container is usually kept frozen after being frozen by the above method.

 [0086] Since the cryopreservation container of the present invention has the above-described configuration, it does not break even at an extremely low temperature such as liquid nitrogen temperature (196 ° C), and the sealing performance at the sealed portion does not deteriorate. It can prevent the contamination and spillage of the contents that liquid refrigerant such as liquid nitrogen does not enter during freezing storage, and exhibits excellent protection performance.

 Furthermore, the cryopreservation container is resistant to temperatures as low as the above-mentioned cryogenic temperatures as long as it is less than the melting point of the adhesive fluorocoagulant used. Therefore, even when the temperature is changed to a room temperature after being placed at the cryogenic temperature, there is no breakage and deterioration of the sealing performance at the sealing portion.

[0087] The mechanism by which the cryopreservation container of the present invention exhibits the above-mentioned excellent effects is not clear. For example, when the above-mentioned adhesive fluorine-containing resin and PI are laminated, (1 ) PI has extremely low temperature resistance that can maintain the shape of the molded body even at extremely low temperatures such as liquid nitrogen temperature. (2) Excellent interlayer adhesion between PI film and adhesive fluororesin layer. ( 3) The PI film and the adhesive fluororesin layer can be directly bonded without using an adhesive, and there is a problem when using an adhesive, that is, the adhesive layer is at a liquid nitrogen temperature. There is no problem of embrittlement and destruction at extremely low temperatures, and no problems due to outgas and elution from the adhesive, and (4 ) A laminated film formed by laminating a PI film and an adhesive fluororesin layer forms a cryopreservation container by thermally fusing the adhesive fluororesin layers to each other. It is considered that the properties of being excellent in adhesion between each other and reliable in sealing properties are synergistically exhibited.

 [0088] The cryopreservation container of the present invention can be suitably used as a cryopreservation container for biological samples.

 Examples of biological samples that can be stored in the cryopreservation container include biological samples derived from humans, biological samples derived from animals or plants other than humans, and biological samples such as viruses and microorganisms.

 [0089] The cryopreservation container of the present invention is a container that can seal biological samples such as blood components, cells, tissues, organs, viruses, bacteria, sperm, eggs, fertilized eggs, and the like.

 [0090] Examples of the blood components include whole blood, red blood cells, white blood cells, plasma, platelets, and platelet-rich plasma. The cells include hematopoietic stem cells, ES cells, mesenchymal stem cells, skeletal mononuclear cells, rare cells such as sperm cells and egg cells, and general cells such as nerve cells, epithelial cells, fibroblasts, etc. Is mentioned. Furthermore, as a biological tissue, as various tissues such as tendons, nerves, ligaments, esophagus, trachea, and spleen, as well as membrane tissues and organs such as mucosal epithelial tissue, corneal epithelial tissue and cultured corneal tissue, Examples include the spleen, heart, lungs, liver and kidneys. Examples of viruses include hepatitis B virus, hepatitis C virus, coronavirus, and mosaic virus. Examples of the bacteria include tuberculosis bacteria, Haemophilus influenzae, Escherichia coli, Staphylococcus aureus, hemolyzed staphylococci, and Klebsiella pneumoniae. In addition, sperm, ovum and fertilized egg are mentioned in the field of infertility treatment, for example. These stored blood or blood components, cells containing rare cells, and other biological tissues are selected according to the purpose of the operator and are not particularly limited.

[0091] Examples of the biological tissue include biological fluids (blood, cerebrospinal fluid, lymph, etc.) and components thereof (red blood cells, white blood cells, platelets, plasma, serum, etc.), and biological tissues (blood vessels, cornea, Meniscus, brain tissue, skin, subcutaneous tissue, epithelial tissue, bone tissue, muscle tissue, etc.), organs (eye, lung, kidney, heart, liver, spleen, spleen, digestive tract, bladder, ovary, testis, etc.), various Cells (such as cord blood, hematopoietic stem cells such as peripheral blood-derived hematopoietic stem cells, bone marrow cells, hepatocytes, spleen cells and brain cells) Various organ cells, nerve cells, sperm, egg cells, fertilized eggs, embryonic stem cells (ES cells), cancer cells for research and treatment, cultured cells, stem cells, embryo cells, etc.).

[0092] Examples of the biological sample include human biological tissues and genetic related substances, as well as biological tissues and genetic related substances of animals including small animals such as small experimental animals; microorganisms, bacteria, and genetic related substances thereof. These include, for example, those used in the research field.

 [0093] Examples of the biological sample also include livestock / animal biological tissues and genetic materials, and examples thereof include those used in the agricultural field such as research, culture, cultivation, and horticulture.

 Examples of the biological sample also include plant seeds, pollen, cultured cells, shoot apical cells, and genetic materials.

 Examples of the biological sample also include biological tissues such as marine algae and fish and genetic related substances, and examples thereof include those used in the field of fisheries such as research.

 Examples of the above-mentioned gene-related substances include DNA, host, vector and the like.

[0094] The biological sample is, for example, for medical use; for research and development in the fields of agriculture, forestry, fisheries, horticulture, etc .; for animal treatment, fertility treatment, etc. in the pet industry and animal industry. It can be used for related purposes, cloning techniques, etc.

As described above, the cryopreservation container of the present invention can be used in various fields such as medicine; research; agriculture such as livestock and horticulture;

 Example

 [0096] Hereinafter, the present invention will be described in detail using examples, but the present invention is not limited to these examples.

 [0097] Examples 1-8: Production of cryopreservation containers

Polytetrafluoroethylene, perfluoro (bulpropyl ether), perfluoro (1, 1, 9, 9-tetrahydro-2, 5-bistrifluoromethyl-3, 6 dioxaxene on one or both sides of the polyimide film 8-component random copolymer film (hereinafter referred to as inner layer fluororesin film and outer layer fluororesin film, respectively) Two-layer and three-layer films were prepared by the heat laminating method. Two sheets of the above film cut to a size of 180 x 100 mm are stacked so that the inner fluororesin film is in contact with each other, and the edge 10 mm is heat-sealed by the heat seal method to form a cryopreservation container. Created. Table 1 shows the film thickness of each film layer.

The copolymer composition ratio of the above three-component random copolymer film is as follows: perfluoro (1, 1, 9, 9—tetrahydride 1, 2, 5-bistrifluoromethyl 1, 3, 6—dioxa 8—nonenor ) For monomer unit 1, the total number of polytetrafluoroethylene monomer units and perfluoro (bulupropyl ether) monomer units was 99. The copolymer composition ratio was measured by ¹⁹ F-NMR.

[0098] Comparative Example 1: Comparative cryopreservation container 1

 A commercially available cryopreservation container (manufactured by BAXER) made of an ethylene-vinyl acetate copolymer was used.

 [0099] Comparative Example 2: Comparative cryopreservation container 2

 A commercially available cryopreservation container made of polyethylene (CharterMed) was used.

 [0100] Synthesis Example 1 (Synthesis of adhesive fluorine resin)

 200L of pure water is placed in a glass-lined autoclave with an internal volume of 820L, the inside of the diameter is sufficiently replaced with nitrogen gas, and then vacuumed to give 113kg of 1-fluoro-1,1-dichloroethane and 95kg of hexafluoropropylene. , 85 g of cyclohexane was charged. Next, perfluoro (1, 1, 5—trihydre port—1—pentene) [CH = CF (CF) H]

 2 2 3

The inside temperature was kept at 35 ° C and the stirring speed was kept at 200 rpm. Further, tetrafluoroethylene was injected into 7.25 kgZcm ² G, and then ethylene was injected into 8 kgZcm ² G.

[0101] Then, by charged with 50 mass _^0/0 methanol solution 1. 9 kg of di-n- propyl Per O carboxymethyl dicarbonate, to initiate polymerization. Since the pressure in the tank decreases as the polymerization progresses

, Tetrafluoroethylene Z ethylene Z hexafluoropropylene mixed gas (molar ratio)

= 39.2: 43.6: 17.3), and the polymerization was continued while maintaining the polymerization pressure at 8 kgZcm¾, and CH = CF (CF) HI lOOg was divided into 20 times during the process. Preparation

 2 2 3

The polymerization was conducted for a total of 32 hours. After polymerization is complete, the contents are recovered, washed with water, and contacted with powder. 95 kg of adhesive fluorocoagulant was obtained.

[0102] The following measurement was performed on the obtained adhesive fluorocoating.

 (1) Monomer unit

¹⁹ F—NMR analysis was performed and measured.

 (2) Number of carbonate groups

Adhesive fluorocobalt powder was compression molded at room temperature to produce a 0.05-0. Infrared absorption spectrum analysis was performed on the resulting film, and the c = o absorbance of the peak [1809cm ^{_ 1} (v;)] attributed to the carbo group in the carbonate group [OC (= O) -O] was measured. did. From the measured values obtained, the number of carbonate groups per IX 10 ⁶ main chain carbon atoms was calculated based on the following formula.

 N = 500AW / ε df

 A: Absorbance of v above

 ε: ν above

(From the model compound, _ε = 170)

 W: Composition average molecular weight calculated from the monomer composition

d: Density of film [g / cm ³ ]

 f: Film thickness [mm] Measured with a micrometer.

 The infrared absorption spectrum analysis described above is based on the Perkin—Elmer FTIR Spectrometer 1760X

Scanning was performed 40 times using (Perkin—Elmer). The absorbance analysis of v is c = o

 Perkin—Elmer Spectrum for Windows (registered trademark) Ver. 1. 4C software.

 (3) Melting point

 Using a differential scanning calorimeter (manufactured by Seiko Co., Ltd.), the temperature was measured at a rate of temperature increase of 10 ° CZ, and the temperature at the maximum value of the melting peak obtained was taken as the melting point.

[0103] The obtained adhesive fluorine resin has a monomer unit of TFEZEtZHFPZ [CH = CF (CF

 2

 H] = 38. 9/45. 9/14. 8/0. 4 and the number of carbonate groups is 1 X 1

twenty three

0 The number was 411 per ⁶ and the melting point was 171.8 ° C.

[0104] Example 9 (1) A T-die is connected to a single-screw extruder with a cylinder diameter of 90 mm using the adhesive fluorine resin obtained from the synthesis example, the cylinder temperature is 170 to 230 ° C, the die temperature is 230 ° C, the screw An adhesive fluororesin film (thickness: 25 m) was molded under the condition of the number of revolutions lOrpm. The adhesive fluororesin film obtained and a polyimide film (product name: Kapton 100H, manufactured by Toray DuPont, thickness: 25 μm) are laminated with a hot roll at a temperature of 250 ° C. Thus, a laminated film (length 20 m × width 200 mm × total thickness 50 πι, hereinafter referred to as a long film) was obtained. The resulting laminated film (fluorine-resin layer thickness: 25 m, polyimide layer thickness: 25 / zm) was cut into a strip of 100 mm in the length direction and 10 mm in the width direction, and its edges Fluorine resin layer and polyimide layer were peeled off using a blade to make a margin, and measured using a Tensilon universal testing machine (Orientec Co., Ltd.) 180 ° peeled at a rate of 25 mmZ, measured at 400 NZm. there were.

 (2) Next, two laminated films cut into 12cm squares from the above long film were stacked with a seal width of 1cm with the adhesive fluorocoagulant layer inside, and the 3 sides were 210 ° in one heat sealer. CX was heat-sealed for 5 seconds. Furthermore, lOOmL pure water is introduced from one side of the opening, and the opening is heat-sealed so that air does not enter, and the internal space filled with the above-mentioned pure water 10 cm long × 10 cm wide × lcm high Created a bag with.

Ten bags were made by this method, and these were immersed in 196 ° C liquid nitrogen for 24 hours and then thawed in 37 ° C warm water. There was no leakage.

From the above, it was considered that the cryopreservation container of the present invention can sufficiently cope with liquid nitrogen immersion.

(3) Laminate two sheets of 12cm square cut from the above long film with a seal width (paste) of lcm, with the adhesive fluororesin layer inside, and 3 sides with heat sealer at 210 ° CX 5 A 25-ml cryopreservation container was prepared by heat-sealing under the conditions of seconds. Table 1 shows the film thicknesses of the polyimide film and adhesive fluorine resin film (inner layer fluorine resin film). Experimental example 1: Freezing test

The cryopreservation containers prepared in Examples 1 to 9 and the cryopreservation containers of Comparative Examples 1 and 2 were each filled with 80 ml of a dimethyl sulfoxide (DMSO) 10% (vZv) aqueous solution, and the air was sufficiently evacuated. Housed in an aluminum case. Dee container stored in aluminum case It was left to stand at 80 ° C for 4 hours in a freezer (freezer) and frozen. The frozen container was then transferred into liquid nitrogen and stored for 1 week. The stored container was also taken out of the aluminum case, and thawed in a 37-40 ° C warm bath, and the container was visually inspected for damage and liquid nitrogen contamination.

 [0106] Table 2 shows the results of the experiment. While Comparative Example 1 was damaged by 20% and Comparative Example 2 was broken by 10%, the cryopreservation container of the present invention had a huge number of tests of 30 pieces, but it was not damaged. I got it.

 [0107] [Table 1]

[0108] [Table 2]

[0109] Experimental Example 2: Freezing test

Cell suspensions were prepared at a concentration of MOLT- 4 cells RPMI1640 medium (manufactured by Invitorogen Co., Ltd.) (the transfer from RIKEN Corporation) about _{^{1. 0 X 10 7 ce lls /}} ml. Made in Example 9 Each of the cryopreservation container and the cryopreservation container of Comparative Example 1 were filled with 80 ml of a cell suspension of dimethylsulfoxide (DMSO) 10% (v / v), fully evacuated, and then made of aluminum. Stored in a case. The container housed in the aluminum case was frozen at 80 ° C for 4 hours using a deep freezer. The frozen container was then transferred into liquid nitrogen and stored for 1 week. The stored container was also taken out of the aluminum case, and thawed in a 37-40 ° C warm bath, and visually inspected the container for damage and liquid nitrogen contamination. Each test was conducted 5 times.

As a result, one of the cryopreservation containers of Comparative Example 1 was damaged, while all five of the cryopreservation containers of the present invention were not damaged.

 Industrial applicability

[0111] The cryopreservation container of the present invention makes it possible to store blood, rare cells, and biological tissues without damage in a cryogenic environment. In addition, since the film thickness is relatively thin, the thermal conductivity to the contents of the cryopreservation container is not reduced. Furthermore, since the sealability by heat sealing after storing the cryopreservation container is very good, it is possible to prevent the inside of the liquid nitrogen from being mixed, and contamination and thawing by bacteria or viruses in the liquid nitrogen. It is possible to prevent the cryopreservation container from rupturing due to the expansion of mixed liquid nitrogen.

Claims

The scope of the claims  [1] A cryopreservation container containing at least an adhesive fluorine-containing polymer film.  2. The cryopreservation container according to claim 1, wherein the adhesive fluorine-containing polymer film comprises an adhesive fluorine-containing polymer having an adhesive site. [3] Adhesive site strength Consists of carbon-carbon double bond, carbonyl group [—C (= 〇)], group or bond having carbonyl group, hydroxyl group, cyano group, sulfonic acid group, and epoxy group The cryopreservation container according to claim 2, which is at least one selected from the group. [4] The adhesive fluorine-containing polymer has a reactive functional group as an adhesive site, and the following (A 4) A cryopreservation container according to claim 3, which is a copolymer obtained by copolymerizing (B) and (B): (A) Fluorine-containing monomer having no reactive functional group  (B) A fluorine-containing monomer having at least one reactive functional group.  [5] Fluorinated monomer power having no reactive functional group The cryopreservation container according to claim 4, which is represented by the following formula (1): [Chemical 1] cx ^! ₂ = cx ' ² (1)  Y (In the formula, X ¹ and X ² are each a hydrogen atom or a halogen atom, and γ is a hydrogen atom. A fluorine atom, a fluorine-containing alkyl group having 1 to 5 carbon atoms, or a fluorine-containing alkylalkyl group having 1 to 5 carbon atoms).  [6] Fluorine-containing monomer power without reactive functional groups Tetrafluoroethylene, vinylidene fluoride, 1,2-difluorochloroethylene, hexafluoropropylene, perfluoro (vinylmethyl) The cryopreservation container according to claim 5, wherein the container is at least one selected from the group consisting of ether) and perfluoro (bulupropyl ether) force.  [7] Fluorinated monomer power having a reactive functional group The cryopreservation container according to any one of claims 4 to 6 represented by the following formula (2): [Chemical 2] c

 c (Wherein x ¹ and x ² are each a hydrogen atom or a halogen atom, z is a hydroxyl group, a carboxyl group, a cyano group, a sulfonic acid group or an epoxy group, and R is the number of carbon atoms f A fluorine-containing alkylene group having 1 to 40 carbon atoms, a fluorine-containing alkylene group having 1 to 40 carbon atoms, or a fluorine-containing alkylene group having at least one ether bond having 1 to 40 carbon atoms).  The cryopreservation capacity according to claim 4, wherein the adhesive fluorine-containing polymer is represented by the following formula (3): ¾5:  [Chemical 3]

(In the formula, X ¹ and X ² are each a hydrogen atom or a halogen atom, and Y ¹ and Y ² are each a hydrogen atom, a fluorine atom, a fluorine-containing alkyl group having 1 to 5 carbon atoms, or a group having 1 to 5 carbon atoms. A fluorine alkoxy group, Z is a hydroxyl group, carboxyl group, cyano group, sulfonic acid group or epoxy group, R is a fluorine-containing alkylene group having 1 to 40 carbon atoms, f  A fluorine-containing oxyalkylene group having 1 to 40 carbon atoms or a fluorine-containing alkylene group having at least one ether bond having 1 to 40 carbon atoms, and (1 + m) Zn force is 2000 [9] The freezing according to claim 3, wherein the adhesive fluorine-containing polymer has a fluorine-containing monomer unit derived from a fluorine-containing monomer and a fluorine-free monomer unit derived from a fluorine-free monomer. Storage container. [10] The fluorine-containing monomer is tetrafluoroethylene, and the fluorine-free monomer is ethylene The cryopreservation container according to claim 9, wherein [11] It is composed of at least an adhesive fluorine-containing polymer film and a laminated film including a film other than the adhesive fluorine-containing polymer film, and the adhesive fluorine-containing polymer film is present on at least one outermost surface. The cryopreservation container according to claim 1. 12. The cryopreservation container according to claim 11, wherein the film other than the adhesive fluorine-containing polymer film is a cold-resistant resin film. [13] The cold-resistant resin is at least 1 selected from the group consisting of ultrahigh molecular weight polyethylene, polyimide, polytetrafluoroethylene, ethylene-tetrafluoroethylene copolymer, and ethylene vinyl acetate copolymer. The cryopreservation container according to claim 12, wherein the container is a cryopreservation container. 14. The cryopreservation container according to claim 13, wherein the cold-resistant rosin is polyimide. [15] A method for producing a cryopreservation container, wherein at least an adhesive fluorine-containing polymer film is formed into a bag shape by heat sealing. [16] A laminated film including at least an adhesive fluorine-containing polymer film and a film other than the adhesive fluorine-containing polymer film, wherein the adhesive fluorine-containing polymer film is present on at least one outermost surface is heated. 16. The method for producing a cryopreservation container according to claim 15, wherein the container is formed into a bag shape with a seal.