Orifice plate and culture method

Abstract

Provided are: a well plate which has excellent adhesion to cells and the like and low drug adsorbability; and a culture method. An orifice plate having a bottom surface member and an orifice-side wall member, in which at least one orifice is formed by joining the bottom surface member and the orifice-side wall member, the bottom surface member and the orifice-side wall member being made of different materials, and the orifice-side wall member having a surface free energy of 18.0-34.0 mN / m.

Description

Technical Field

[0001] One embodiment of the present invention relates to a well plate or a culture method. Background Art

[0002] In the cultivation of cells, tissues and organs (hereinafter also collectively referred to as "cells, etc.") in medical treatment or drug development such as drug screening, an orifice plate having one or more holes consisting of a bottom surface and a side wall is widely used. The orifice plate is generally formed in a manner that the bottom surface and the side wall are integrated. In addition, as a material for the orifice plate, polystyrene is widely used from the viewpoint of transparency and cost.

[0003] When cells and the like are cultured using a well plate, a culture medium and cells and the like are placed in the wells, and the cells and the like are cultured on the bottom surface of the wells. When the culture medium is placed in the wells, the culture medium is usually allowed to flow into the wells along the side walls of the wells so as not to damage the cells and the like on the bottom surface of the wells.

[0004] However, it is known that polystyrene easily adsorbs drugs and proteins and has high drug adsorption. Therefore, when using an integrally formed polystyrene well plate, when the culture medium is allowed to flow along the well side wall, the drugs and proteins in the culture medium are adsorbed by the polystyrene constituting the well side wall, and there is a problem that the drugs and proteins cannot be fully distributed to the cells in the well.

[0005] Patent Document 1 discloses a plastic molded product used in the field of biotechnology as a plastic molded product to which DNA and protein are difficult to adhere. The plastic molded product is composed of a molded product molded using a plastic material, wherein the plastic material is based on a thermoplastic plastic and a silicone copolymer resin is added to the thermoplastic plastic and mixed.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: Japanese Patent Application Publication No. 2022-064159 Summary of the invention

[0009] Problems to be solved by the invention

[0010] The plastic moldings recorded in patent document 1 are that the water-repellent and oil-repellent effects of the plastic surface are improved, and can show the same function as when coated with organic silicon raw materials. According to the research of the inventors of the present invention, the drug adsorption of the plastic moldings recorded in patent document 1 is lower than that of polystyrene orifice plate, although the drug adsorption is excellent, the water-repellent and oil-repellent effects are strong, and it is not only easy to repel drugs, but also easy to repel cells, culture medium, coating agents, etc. That is, the plastic moldings recorded in patent document 1 leave room for improvement in the adhesion of cells, etc.

[0011] Therefore, one embodiment of the present invention provides a well plate and a culture method having excellent adhesion to cells and the like and low drug adsorption.

[0012] Means for solving problems

[0013] The inventors of the present invention have conducted intensive studies to solve the above-mentioned problems. As a result, they have found that the above-mentioned problems can be solved by having the following configuration, and have completed the present invention.

[0014] Embodiments of the present invention relate to, for example, the following [1] to

[14] .

[0015] [1] A perforated plate having a bottom member and a hole side wall member, wherein at least one hole is formed by joining the bottom member and the hole side wall member, the bottom member and the hole side wall member are made of different materials, and the surface free energy of the hole side wall member is 18.0 to 34.0 mN / m.

[0016] [2] The orifice plate according to [1], wherein the orifice side wall member contains a thermoplastic resin.

[0017] [3] The orifice plate according to [2], wherein the thermoplastic resin is a polyolefin-based resin.

[0018] [4] The orifice plate according to [3], wherein the polyolefin resin is at least one selected from a cyclic olefin copolymer (α), a 4-methyl-1-pentene polymer (β), a propylene polymer (γ) and a cyclic olefin polymer (δ) (excluding the cyclic olefin copolymer (α)).

[0019] [5] The orifice plate according to [4], wherein the orifice sidewall member further comprises a silanized polyolefin (ε).

[0020] [6] The orifice plate according to any one of [1] to [5], wherein the bottom surface member has a glass transition temperature of -40°C or higher as measured in accordance with JIS-K7121:1987.

[0021] [7] The orifice plate according to any one of [1] to [6], wherein the total light transmittance of the bottom surface member measured in accordance with JIS-K-7361-1 is 80.0% or more.

[0022] [8] The orifice plate according to any one of [1] to [7], wherein the bottom surface member has a thickness of 20 to 500 μm.

[0023] [9] The well plate according to any one of [1] to [8], wherein the culture surface of the bottom member is coated with at least one coating agent selected from natural polymer materials, synthetic polymer materials and inorganic materials.

[0024]

[10] The well plate according to any one of [1] to [9], wherein the water contact angle of the culture surface of the bottom member is 50 to 100°.

[0025]

[11] The orifice plate according to any one of [1] to

[10] , wherein the glass transition temperature of the orifice side wall member measured by solid viscoelastic temperature dispersion measurement is 30° C. or higher.

[0026]

[12] The orifice plate according to any one of [1] to

[11] , wherein the glass transition temperature of the orifice side wall member measured by solid viscoelastic temperature dispersion measurement is 120° C. or higher.

[0027]

[13] The orifice plate according to any one of [1] to

[12] , wherein the surface free energy of the orifice side wall member is 18.0 to 24.3 mN / m.

[0028]

[14] A method for culturing cells, tissues or organs, comprising the step of culturing cells, tissues or organs using the well plate according to any one of [1] to

[13] .

[0029] Effects of the Invention

[0030] According to one embodiment of the present invention, a well plate and a culture method having excellent adhesion to cells and the like and low drug adsorption can be provided. DETAILED DESCRIPTION

[0031] Next, the present invention will be described in detail.

[0032] In the present disclosure, the description "XX or more and YY or less" and "XX to YY" indicating a numerical range means a numerical range including the lower limit and the upper limit as endpoints unless otherwise specified.

[0033] Moreover, when describing a numerical range in stages, the upper limit and the lower limit of each numerical range can be arbitrarily combined.

[0034] <Orifice Plate>

[0035] An orifice plate of one embodiment of the present invention has a bottom surface member and a hole side wall member, at least one hole is formed by the bottom surface member and the hole side wall member being joined, the bottom surface member and the hole side wall member being made of different materials, and the surface free energy of the hole side wall member being 18.0 to 34.0 mN / m. Hereinafter, the orifice plate is also referred to as an orifice plate (X).

[0036] The well side wall member and the bottom surface member of the well plate (X) are joined so that cells or the culture medium in the wells do not leak out. The joining may be any of mechanical joining, material joining, and adhesive joining.

[0037] The well plate (X) is a plate having at least one well. The well plate (X) is preferably a multi-well plate having a plurality of wells such as 6 wells, 12 wells, 24 wells, 48 ​​wells, 96 wells, 384 wells, 1536 wells, etc.

[0038] The orifice plate (X) may have a cover covering the upper portion of the plate.

[0039] The well plate (X) can be used in a culture method having a step of culturing cells, tissues or organs. The cells, tissues and organs are not particularly limited, and the well plate (X) can be used for culturing known cells, tissues and organs.

[0040] The well plate (X) is preferably an adhesive material because cells etc. have excellent adhesion. The cells etc. are preferably cells, and more preferably adhesive cells, because they are suitable for culture on the culture surface of the well plate (X).

[0041] There is no particular restriction on the manufacturing method of the orifice plate (X), and there is no particular restriction on the equipment used for manufacturing. The orifice plate (X) can be obtained, for example, by forming the hole side wall member into a desired shape by extrusion molding, solution casting molding, injection molding, blow molding, etc., and then joining it with the bottom surface member. There is no particular restriction on the joining method, and examples thereof include: mechanical joining such as clamping with an elastic spring member or fastening with a threaded member; material joining such as welding using heat, ultrasound, laser, high-frequency medium, etc.; adhesive joining using an adhesive or bonding agent.

[0042] <Bottom member>

[0043] The bottom surface member is a member constituting the bottom surface of the orifice plate (X), and forms at least one orifice by being joined with the orifice side wall member. When the bottom surface member forms at least one orifice by being joined with the orifice side wall member, the bottom surface member constitutes the bottom surface of the orifice.

[0044] The material constituting the bottom surface member can be any material different from the material constituting the hole side wall member, without any particular restrictions, and known materials (such as resins such as thermosetting resins and thermoplastic resins, glass, etc.) can be used. Specifically, as the material constituting the hole side wall member, the bottom surface member can be constituted by the same material as the material described later. However, it is necessary to be a material different from the material constituting the hole side wall member. It should be noted that different materials refer to: the main material constituting the bottom surface member (the material constituting the member contains the most material on a mass basis) is different from the main material constituting the hole side wall member. In the case where the main material is a (co)polymer, different materials refer to: excluding the situation where the types and proportions of the structural units constituting the polymer are the same. For example, even if the material constituting the bottom surface member and the material constituting the hole side wall member contain a common structural unit, as long as the ratio of the structural unit is different between the material constituting the bottom surface member and the material constituting the hole side wall member, the material constituting the bottom surface member and the material constituting the hole side wall member are also different.

[0045] The material constituting the bottom surface member preferably comprises a thermoplastic resin or glass, and more preferably comprises a polyolefin resin. The polyolefin resin is preferably at least one selected from a cyclic olefin copolymer (α), a 4-methyl-1-pentene polymer (β), a propylene polymer (γ), and a cyclic olefin polymer (δ) (wherein the cyclic olefin copolymer (α) is not included), more preferably a 4-methyl-1-pentene polymer (β), a propylene polymer (γ), and further preferably a 4-methyl-1-pentene polymer (β).

[0046] The glass transition temperature (Tg) of the bottom surface member measured in accordance with JIS-K7121: 1987 is preferably -40°C or higher, more preferably -20°C or higher. The upper limit of Tg is not particularly limited, and can be, for example, 600°C or lower.

[0047] When the Tg of the bottom surface member is within the above range, the bottom surface member has excellent flexibility and strength, and has excellent low drug adsorption properties.

[0048] The total light transmittance of the bottom member measured in accordance with JIS-K-7361-1 is preferably 80.0% or more, more preferably 90.0% or more. The upper limit of the total light transmittance is not particularly limited, and may be, for example, 100.0% or less or less.

[0049] When the total light transmittance of the bottom surface member is within the above range, the bottom surface member has excellent transparency, and cells and the like can be easily observed both with the naked eye and under a microscope.

[0050] The thickness of the bottom member is preferably 20 to 500 μm, more preferably 40 to 300 μm. If the thickness of the bottom member is within the above range, the bottom surface of the culture container will not bend when culturing cells, and a suitable culture container can be manufactured.

[0051] The shape of the bottom surface member is not particularly limited as long as at least one hole can be formed by joining with the hole side wall member, and is preferably in the shape of a plate, a film, or a sheet.

[0052] The shape of the inner side of the hole of the bottom surface member can include a flat bottom (F bottom), a round bottom (U bottom), a conical bottom (V bottom), a flat bottom + a curved edge, etc. In the case of processing into a round bottom (U bottom), a conical bottom (V bottom), a flat bottom + a curved edge, etc., a plate, a membrane or a sheet can also be made in advance, and a second process such as vacuum forming or pressure forming can be performed to make it. The shape of the inner side of the hole of the bottom surface member can be selected according to the purpose of culture. When cells are cultured in two dimensions, a flat bottom (F bottom) is usually preferred, and when cells are cultured in three dimensions, a round bottom (U bottom) or a conical bottom (V bottom) is usually preferred.

[0053] The manufacturing method of the bottom surface member is not particularly limited, and the equipment used for manufacturing is also not limited. The plate, film, sheet or other molded product as the bottom surface member can be obtained by directly molding the material constituting the bottom surface member using methods such as extrusion molding, solution casting molding, injection molding, blow molding, etc.

[0054] As a method for forming the film or sheet, specifically, for example, a common inflation method, a T-die extrusion method, etc. are used. The production is usually carried out by heating. In the case of the T-die extrusion method, the extrusion temperature varies depending on the type of material constituting the bottom surface member, and is preferably 100°C to 400°C, and particularly preferably 150°C to 300°C. In addition, the roll temperature is preferably 20°C to 100°C, and particularly preferably 30°C to 90°C.

[0055] In addition, the film or sheet can also be manufactured by a solution casting method, in which the material constituting the bottom surface member is dissolved in a solvent and made to flow over a resin or metal, and is slowly dried while being leveled and filmized (sheetized). The solvent used is not particularly limited, and hydrocarbon solvents such as cyclohexane, hexane, decane, and toluene can be used. In addition, considering the solubility and drying efficiency of the material, the solvent can be mixed with more than two kinds. The polymer solution can be applied by methods such as bench coating, spin coating, dip coating, die coating, spray coating, rod coating, roller coating, curtain flow coating, dried, peeled off, and thus processed into a film or sheet.

[0056] At least the culture surface of the bottom surface member, in particular the bottom surface member, is preferably surface treated. Here, "culture surface" refers to the surface inside the hole of the bottom surface member, the surface that the cells etc. contact when using a well plate to culture cells etc. or the surface that is planned to contact.

[0057] By surface treating at least the culture surface of the bottom surface member, it can be made into a state where cells etc. can be easily adhered, and cells etc. can be easily proliferated uniformly. In addition, the wettability of the culture surface is improved, and at least one coating agent selected from natural polymer materials, synthetic polymer materials and inorganic materials can be easily applied on the culture surface. The coating will be described later.

[0058] Surface treatment is preferably performed before joining the bottom surface member to the hole side wall member.

[0059] As surface treatment, for example, hydrophilization treatment can be mentioned. The method used in the hydrophilization treatment is not particularly limited, and for example, corona treatment, plasma treatment, ozone treatment, ultraviolet treatment, chemical vapor deposition, etching, or addition of specific functional groups such as hydroxyl, amino, sulfonic group, thiol group, carboxyl, etc., the treatment of specific functional groups such as silane coupling, the surface roughening using oxidants, etc. can be mentioned. Wherein, in order to improve the wettability of the bottom surface member, cells can be effectively cultured, etc., it is preferred to carry out surface hydrophilization treatments such as ultraviolet treatment, corona treatment, plasma treatment or ozone treatment. These surface treatments can be carried out alone, or two or more can be combined.

[0060] In these surface treatments, owing to being able to make it become the state of easy bonding surface, the culture surface of the preferred bottom surface member at least is the culture surface treated with plasma.As plasma treatment, vacuum plasma treatment, normal pressure plasma treatment and high pressure plasma treatment can be enumerated.When carrying out plasma treatment, as accompanying gas, use nitrogen, hydrogen, helium, oxygen, argon etc., preferably select at least one gas selected from nitrogen, helium, argon.

[0061] The culture surface of the bottom member is preferably a culture surface coated with at least one coating agent selected from the group consisting of natural polymer materials, synthetic polymer materials, and inorganic materials.

[0062] There is no particular limitation on the coating agent. Examples of natural polymer materials include glycosaminoglycans such as collagen, gelatin, alginate, hyaluronic acid, chondroitin sulfate, fibronectin, laminin, fibrinogen, osteopontin, tenascin, vitronectin, thrombospondin, agarose, elastin, keratin, chitosan, fibrin, silk protein, and sugars. Examples of synthetic polymer materials include polyglycolic acid, polylactic acid, polyethylene glycol, polycaprolactone, synthetic peptides, synthetic proteins, polyhydroxyethyl methacrylate, and polyethyleneimine. Examples of inorganic materials include β-tricalcium phosphate and calcium carbonate.

[0063] In addition, as the coating agent, vitrigel obtained by vitrifying and then hydrating a hydrogel of a conventional extracellular matrix component, etc. For example, collagen vitrigel consisting of a high-density collagen fiber network made of collagen, which is one of the extracellular matrix components, can also be mentioned.

[0064] From the viewpoints of improving the adhesion or proliferation of cells, etc., and maintaining the functions of cells, etc. for a longer period of time, the coating agent is preferably a protein or peptide such as collagen, gelatin, laminin, polylysine, etc., more preferably laminin, collagen or polylysine, and further preferably collagen. These coating agents may be a single species or a combination of two or more species.

[0065] The bottom surface member is subjected to the above-mentioned coating treatment so that even when it is washed with physiological saline after the coating treatment or placed in a culture environment of cells, the well plate will not deform, discolor or peel off, and can maintain a stable initial state and be used as a culture container, so it is preferred.

[0066] The water contact angle of the culture surface of the bottom member is preferably 30° or more, more preferably 35° or more, further preferably 40° or more, particularly preferably 45° or more, and further preferably 50° or more. The water contact angle is preferably 120° or less, more preferably 110° or less, further preferably 100° or less, and particularly preferably 84° or less or 70° or less. The upper and lower limits may be combined arbitrarily.

[0067] By adjusting the water contact angle of the bottom surface member to the above range, for example, cells etc. become easy to adhere to the bottom surface member, and cells etc. are easy to proliferate evenly on the bottom surface member. In addition, the natural polymer material, synthetic polymer material or inorganic material is evenly coated on the bottom surface member to make it easy to adhere. In addition, even after coating, when washed with physiological saline or placed in a cell culture environment, the natural polymer material, synthetic polymer material or inorganic material will not peel off, and can maintain a stable initial state for cell culture.

[0068] The water contact angle is measured by a sessile drop method. The details of the measurement method will be described later.

[0069] <Hole side wall member>

[0070] The hole side wall member is a member constituting a side wall of a hole, and is joined with the bottom surface member to form at least one hole.

[0071] The surface free energy of the hole sidewall member is 18.0 to 34.0 mN / m. From the viewpoint of drug adsorption and processability, the surface free energy of the hole sidewall member is preferably 20.0 mN / m or more, more preferably 22.0 mN / m or more, and preferably 30.0 mN / m or less, more preferably 26.0 mN / m or less, and further preferably 24.3 mN / m or less. These upper and lower limits may be arbitrarily combined.

[0072] If the surface free energy of the hole side wall member is less than 18.0 mN / m, sufficient bonding strength with the bottom surface member cannot be obtained, and if it exceeds 34.0 mN / m, sufficient drug adsorption cannot be obtained.

[0073] The glass transition temperature (Tg) of the hole sidewall member measured by solid viscoelastic temperature dispersion measurement is preferably -10°C or higher, more preferably 30°C or higher, and further preferably 120°C or higher. The upper limit of Tg is not particularly limited, and can be, for example, 300°C or lower.

[0074] When the Tg of the hole sidewall member is within the above range, the flexibility and strength are excellent. In addition, under the normal temperature of 25°C to 40°C when culturing cells, the molecules of the hole sidewall member are not easy to move, and as a result, it is not easy to adsorb the drug, so it is preferred.

[0075] The material constituting the hole side wall member is not particularly limited as long as the material has a surface free energy within the above range and is different from that of the bottom surface member, and a known thermosetting resin, thermoplastic resin, etc. can be used.

[0076] The hole side wall member preferably comprises a thermoplastic resin. The thermoplastic resin is not particularly limited, and examples thereof include: polyolefin resins such as polyethylene, polypropylene, cyclic olefin polymers, polymethylpentene resins, and norbornene polymers; polymethacrylic resins such as polymethyl methacrylate resins; polyacrylic resins such as polymethyl acrylate resins; polyvinyl acetal resins; polyvinyl butyral resins; polyvinyl formal resins; polycarbonate resins; polyetheretherketone resins; polyetherketone resins; polyester resins; polyamide resins such as nylon-6, nylon-66, and poly(m-xylylene adipamide); Polyamide-imide resin; polyimide resin; polyetherimide resin; styrene-based elastomer; polyolefin-based elastomer; polyurethane-based elastomer; polyester-based elastomer; polyamide-based elastomer; polytetrafluoroethylene resin; ethylene tetrafluoroethylene copolymer; polyvinylidene fluoride resin; polyvinyl fluoride resin; thermoplastic polyimide resin; polyvinylidene chloride resin; polyvinyl chloride resin; polyvinyl acetate resin; polyphenylene ether resin, polyphenylene sulfide resin and other polyphenylene resins; polysulfone resin; polylactic acid resin; polyethersulfone resin; polyacrylonitrile resin, styrene-acrylonitrile copolymer resin, etc.

[0077] The thermoplastic resin is preferably a polyolefin resin, and the polyolefin resin is preferably at least one selected from a cyclic olefin copolymer (α), a 4-methyl-1-pentene polymer (β), a propylene polymer (γ), and a cyclic olefin polymer (δ) (excluding the cyclic olefin copolymer (α)).

[0078] (Cyclic olefin copolymer (α))

[0079] Hereinafter, the cyclic olefin-based copolymer (α) will be described, but the cyclic olefin-based copolymer (α) is not limited to the following embodiment.

[0080] The cyclic olefin-based copolymer (α) is a copolymer of an α-olefin (including ethylene) and a cyclic olefin.

[0081] The cyclic olefin compound constituting the cyclic olefin copolymer (α) is not particularly limited, and examples thereof include cyclic olefin monomers described in paragraphs 0037 to 0063 of International Publication No. 2006 / 118261.

[0082] From the viewpoint of maintaining the transparency of the obtained component well and further improving the heat resistance or improving the moldability, the cyclic olefin-based copolymer (α) preferably has at least one structural unit (a) derived from olefin represented by the following formula (I), and at least one structural unit (b) derived from cyclic olefin selected from the group consisting of a structural unit represented by the following formula (II), a structural unit represented by the following formula (III) and a structural unit represented by the following formula (IV).

[0083] [Chemistry 1]

[0084]

[0085] In the above formula (I), R 300 It represents a hydrogen atom or a linear or branched hydrocarbon group having 1 to 29 carbon atoms.

[0086] [Chemistry 2]

[0087]

[0088] In the above (II), u is 0 or 1, v is 0 or a positive integer, preferably an integer of 0 or more and 2 or less, more preferably 0 or 1, w is 0 or 1, R 61 ~R 78 and R a1 and R b1 may be the same as or different from each other, and are a hydrogen atom, a halogen atom, an alkyl group having 1 to 20 carbon atoms, a halogenated alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 15 carbon atoms, or an aromatic hydrocarbon group having 6 to 20 carbon atoms, R 75 ~R 78 They may be bonded to each other to form a single ring or multiple rings.

[0089] [Chemistry 3]

[0090]

[0091] In the above formula (III), x and d are integers of 0 or more, preferably integers of 0 or more and 2 or less, more preferably 0 or 1, y and z are 0, 1 or 2, R 81 ~R 99 may be the same as or different from each other, and are a hydrogen atom, a halogen atom, an alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 15 carbon atoms, i.e., an aliphatic hydrocarbon group, an aromatic hydrocarbon group having 6 to 20 carbon atoms, or an alkoxy group, R 89 and R 90 The carbon atom bonded to R 93 The bonded carbon atom or R 91 The carbon atom to which the bond is attached may be directly bonded or bonded via an alkylene group having 1 to 3 carbon atoms. When y=z=0, R 95 With R 92 or R 95 With R 99 They may be bonded to each other to form a monocyclic or polycyclic aromatic ring.

[0092] [Chemistry 4]

[0093]

[0094] In the above formula (IV), R100 , R 101 They may be the same as or different from each other, and represent a hydrogen atom or a hydrocarbon group having 1 to 5 carbon atoms, and f is 1≤f≤18.

[0095] The olefin monomer as one of the copolymerization raw materials of the cyclic olefin copolymer (α) is a monomer which forms the structural unit represented by the above formula (I) by addition copolymerization. Specifically, an olefin monomer represented by the following formula (Ia) corresponding to the above formula (I) is used.

[0096] [Chemistry 5]

[0097]

[0098] In the above formula (Ia), R 300 It represents a hydrogen atom or a linear or branched hydrocarbon group having 1 to 29 carbon atoms.

[0099] As the olefin monomer shown in the above-mentioned formula (Ia), for example, ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 3-methyl-1-butene, 3-methyl-1-pentene, 3-ethyl-1-pentene, 4-methyl-1-pentene, 4-methyl-1-hexene, 4,4-dimethyl-1-hexene, 4,4-dimethyl-1-pentene, 4-ethyl-1-hexene, 3-ethyl-1-hexene, 1-octene, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, 1-eicosene, etc. From the viewpoint of obtaining an orifice plate with more excellent heat resistance, mechanical properties and optical properties, among them, ethylene and propylene are preferred, and ethylene is particularly preferred. The olefin monomer shown in the above-mentioned formula (Ia) can use more than 2 kinds.

[0100] When the total amount of the structural units constituting the cyclic olefin-based copolymer (α) is set to 100 mol%, the proportion of the structural units (a) derived from olefins is preferably from 5 mol% to 95 mol%, more preferably from 20 mol% to 90 mol%, further preferably from 40 mol% to 80 mol%, particularly preferably from 50 mol% to 70 mol%.

[0101] It should be noted that the proportion of the structural unit (a) derived from olefins can be 13 The measurement was performed by C-NMR.

[0102] The cyclic olefin monomer (b) as one of the copolymerization raw materials of the cyclic olefin copolymer (α) is a monomer which is subjected to addition copolymerization to form a structural unit (b) derived from a cyclic olefin represented by the above formula (II), the above formula (III) or the above formula (IV). Specifically, the cyclic olefin monomer (b) represented by the formula (IIa), (IIIa) and (IVa) corresponding to the above formula (II), the above formula (III) and the above formula (IV), respectively, is used.

[0103] [Chemistry 6]

[0104]

[0105] In the above formula (IIa), u is 0 or 1, v is 0 or a positive integer, preferably an integer of 0 or more and 2 or less, more preferably 0 or 1, w is 0 or 1, and R 61 ~R 78 and R a1 and R b1 may be the same as or different from each other, and are a hydrogen atom, a halogen atom, an alkyl group having 1 to 20 carbon atoms, a haloalkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 15 carbon atoms, or an aromatic hydrocarbon group having 6 to 20 carbon atoms, R 75 ~R 78 They may be bonded to each other to form a single ring or multiple rings.

[0106] [Chemistry 7]

[0107]

[0108] In the above formula (IIIa), x and d are integers of 0 or more, preferably integers of 0 or more and 2 or less, more preferably 0 or 1, y and z are 0, 1 or 2, R 81 ~R 99 may be the same as or different from each other, and are a hydrogen atom, a halogen atom, an alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 15 carbon atoms, i.e., an aliphatic hydrocarbon group, an aromatic hydrocarbon group having 6 to 20 carbon atoms, or an alkoxy group, R 89 and R 90 The carbon atom bonded to R 93 The bonded carbon atom or R 91 The carbon atom to which the bond is attached may be directly bonded or bonded via an alkylene group having 1 to 3 carbon atoms. When y=z=0, R 95 With R 92 or R 95 With R 99 They may be bonded to each other to form a monocyclic or polycyclic aromatic ring.

[0109] [Chemistry 8]

[0110]

[0111] In the above formula (IVa), R 100 , R 101 They may be the same as or different from each other, and represent a hydrogen atom or a hydrocarbon group having 1 to 5 carbon atoms, and f is 1≤f≤18.

[0112] By using the olefin monomer represented by the above formula (Ia) and the cyclic olefin monomer (b) represented by the formula (IIa), (IIIa) or (IVa) as a copolymerization component, the solubility of the cyclic olefin copolymer (α) in the solvent is further improved, so that the moldability becomes good and the product yield is improved.

[0113] As specific examples of the cyclic olefin monomer (b) represented by the formula (IIa), (IIIa) or (IVa), the compounds described in paragraphs 0037 to 0063 of International Publication No. 2006 / 118261 can be used.

[0114] Specific examples include bicyclo-2-heptene derivatives (bicyclohept-2-ene derivatives), tricyclo-3-decene derivatives, tricyclo-3-undecene derivatives, tetracyclo-3-dodecene derivatives, pentacyclo-4-pentadecenes derivatives, pentacyclopentadecadiene derivatives, pentacyclo-3-pentadecenes derivatives, pentacyclo-4-hexadecene derivatives, pentacyclo-3-hexadecene derivatives, hexacyclo-4-heptadecenes derivatives, heptacyclo-5-eicosene derivatives. Biological substances, heptacyclic-4-eicosene derivatives, heptacyclic-5-heneicosene derivatives, octacyclic-5-hexadecene derivatives, nonacyclic-5-pentadecene derivatives, nonacyclic-6-hexadecene derivatives, cyclopentadiene-acenaphthylene adducts, 1,4-endomethyl-1,4,4a,9a-tetrahydrofluorene derivatives, 1,4-endomethyl-1,4,4a,5,10,10a-hexahydroanthracene derivatives, and cycloalkylene derivatives having 3 to 20 carbon atoms.

[0115] Among the cyclic olefin monomers (b) represented by the formula (IIa), (IIIa) or (IVa), the cyclic olefin represented by the formula (IIa) is preferred.

[0116] As the cyclic olefin monomer (b) represented by the above formula (IIa), bicyclo[2.2.1]-2-heptene (also called norbornene), tetracyclo[6.2.1.1 3,6 .0 2,7 ] dodec-4-ene (also known as tetracyclo[4.4.0.1 2,5 .1 7,10 ]-3-dodecene or tetracyclododecene), more preferably tetracyclo[6.2.1.1 3,6 .0 2,7These cyclic olefins have a rigid ring structure and thus have the advantage of being easy to maintain the elastic modulus of the copolymer and the obtained member.

[0117] When the total amount of the structural units constituting the cyclic olefin copolymer (α) is set to 100 mol%, the proportion of the structural units (b) derived from the cyclic olefin monomer (b) is preferably 5 mol% to 95 mol%, more preferably 10 mol% to 80 mol%, further preferably 20 mol% to 60 mol%, particularly preferably 30 mol% to 50 mol%.

[0118] The copolymerization type of the cyclic olefin copolymer (α) is not particularly limited, and examples thereof include random copolymers and block copolymers. In the present embodiment, a random copolymer is preferably used as the cyclic olefin copolymer (α) from the viewpoint of excellent transparency.

[0119] The cyclic olefin copolymer (α) is preferably ethylene and tetracyclo[4.4.0.1 2,5 .1 7,10 ]-3-dodecene random copolymers and ethylene and bicyclo[2.2.1]-2-heptene random copolymers, more preferably ethylene and tetracyclo[4.4.0.1 2 ,5 .1 7,10 ]-random copolymer of 3-dodecene.

[0120] The cyclic olefin-based copolymer (α) may be used alone or in combination of two or more.

[0121] The cyclic olefin copolymer (α) can be produced by selecting appropriate conditions according to the methods of, for example, JP-A-60-168708, JP-A-61-120816, JP-A-61-115912, JP-A-61-115916, JP-A-61-271308, JP-A-61-272216, JP-A-62-252406, and JP-A-62-252407.

[0122] As the cyclic olefin copolymer (α), a commercially available product can be used. Examples of commercially available cyclic olefin copolymers (α) include APEL series manufactured by Mitsui Chemicals, Inc. (e.g., APEL6015T, etc.), COXEC series manufactured by Kurabo Co., Ltd. (e.g., COXEC TCS-1), and TOPAS series manufactured by Polyplastics Corporation.

[0123] (4-Methyl-1-pentene polymer (β))

[0124] Hereinafter, the 4-methyl-1-pentene polymer (β) will be described, but the 4-methyl-1-pentene polymer (β) is not limited to the following embodiment.

[0125] The 4-methyl-1-pentene polymer (β) includes a homopolymer of 4-methyl-1-pentene and a copolymer of 4-methyl-1-pentene and other monomers. In addition, the homopolymer of 4-methyl-1-pentene is hereinafter also referred to as a 4-methyl-1-pentene homopolymer (β1).

[0126] The copolymer of 4-methyl-1-pentene and other monomers may be any of a random copolymer, an alternating copolymer, a block copolymer and a graft copolymer. The copolymer of 4-methyl-1-pentene and other monomers is preferably a copolymer (β2) of 4-methyl-1-pentene and at least one olefin selected from ethylene and α-olefins having 3 to 20 carbon atoms (excluding 4-methyl-1-pentene) because the obtained member has high strength (not easy to break or crack) and little deflection.

[0127] The 4-methyl-1-pentene polymer (β) is preferably at least one polymer selected from a 4-methyl-1-pentene homopolymer (β1) and a copolymer (β2) of 4-methyl-1-pentene and at least one olefin selected from ethylene and α-olefins having 3 to 20 carbon atoms (excluding 4-methyl-1-pentene), and more preferably a copolymer (β2) of 4-methyl-1-pentene and at least one olefin selected from ethylene and α-olefins having 3 to 20 carbon atoms (excluding 4-methyl-1-pentene).

[0128] As the olefin, for example, ethylene, propylene, 1-butene, 1-hexene, 1-heptene, 1-octene, 1-decene, 1-tetradecene, 1-hexadecene, 1-heptadecene, 1-octadecene and 1-eicosene can be cited. The olefin can be appropriately selected according to the physical properties required for the culture material. For example, as the olefin, from the viewpoint of moderate oxygen permeability and excellent rigidity, α-olefins with 8 to 18 carbon atoms are preferred, and at least one of 1-octene, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-heptadecene and 1-octadecene is more preferably selected. If the number of carbon atoms of the olefin is within the above range, the film-forming processability of the polymer becomes better, and as a result, there is a tendency that it is not easy to produce a poor appearance caused by cracks or ruptures at the end when demolding from a roller or a mold during molding. Therefore, the defective product incidence rate of the obtained component becomes low.

[0129] The olefin may contain only one kind, or two or more kinds may be combined. From the viewpoint of the strength of the material, the number of carbon atoms is preferably 10 or more. When two or more different α-olefins are combined, it is particularly preferred to combine at least one selected from 1-tetradecene and 1-hexadecene with at least one selected from 1-heptadecene and 1-octadecene.

[0130] The content of the structural unit derived from 4-methyl-1-pentene is preferably 60 to 100 mol%, more preferably 80 to 98 mol%. In addition, the content of the structural unit derived from at least one olefin selected from ethylene and α-olefins having 3 to 20 carbon atoms (excluding 4-methyl-1-pentene) is preferably 0 to 40 mol%, more preferably 2 to 20 mol%. It should be noted that the content of the structural unit is based on the amount of all repeating structural units as 100 mol%. If the content of the structural unit is within the above range, a homogeneous component with excellent processability can be obtained, and the balance between toughness and strength is good, so the deflection is also reduced.

[0131] The 4-methyl-1-pentene polymer (β) may also have structural units other than structural units derived from 4-methyl-1-pentene and structural units derived from the olefins (hereinafter also referred to as "other structural units") within a range that does not impair the effects of the present invention. The content of other structural units is, for example, 0 to 10.0 mol%. When the 4-methyl-1-pentene polymer (β) has other structural units, the other structural units may be one type or two or more types.

[0132] Examples of monomers from which other structural units are derived include cyclic olefins, aromatic vinyl compounds, conjugated dienes, non-conjugated polyenes, functional vinyl compounds, hydroxyl-containing olefins, and halogenated olefins. Examples of cyclic olefins, aromatic vinyl compounds, conjugated dienes, non-conjugated polyenes, functional vinyl compounds, hydroxyl-containing olefins, and halogenated olefins include compounds described in paragraphs

[0035] to

[0041] of Japanese Patent Application Laid-Open No. 2013-169685.

[0133] The 4-methyl-1-pentene polymer (β) may be used alone or in combination of two or more.

[0134] As the 4-methyl-1-pentene polymer (β), a commercially available product can be used. As a commercially available 4-methyl-1-pentene polymer, TPX series manufactured by Mitsui Chemicals, Inc. can be used.

[0135] (Propylene polymer (γ))

[0136] Hereinafter, the propylene-based polymer (γ) will be described, but the propylene-based polymer (γ) is not limited to the following embodiment.

[0137] The propylene-based polymer (γ) refers to a polymer containing a structural unit derived from propylene in a ratio of more than 50 mol% and 100 mol% or less in the polymer.

[0138] Examples of the propylene-based polymer (γ) include a propylene homopolymer (γ1) and a copolymer (γ2) of propylene and at least one α-olefin other than propylene having 2 to 20 carbon atoms. Examples of the α-olefin other than propylene having 2 to 20 carbon atoms include ethylene, 1-butene, 1-pentene, 1-hexene, 4-methyl-1-pentene, 1-octene, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, 1-eicosene, and the like, and ethylene or an α-olefin having 4 to 10 carbon atoms is preferred.

[0139] The copolymer (γ2) of propylene and at least one α-olefin having 2 to 20 carbon atoms other than propylene may be a random copolymer or a block copolymer. The structural units derived from the α-olefin may be contained in the copolymer (γ2) of propylene and at least one α-olefin having 2 to 20 carbon atoms other than propylene in a ratio of preferably 35 mol% or less, more preferably 30 mol% or less.

[0140] The propylene-based polymer (γ) may be either isotactic polypropylene or syndiotactic polypropylene.

[0141] As the propylene-based polymer (γ), a commercially available product can be used. As the commercially available propylene-based polymer (γ), NOVATEC BC03C manufactured by Nippon Polypro Co., Ltd., F107 manufactured by Prime Polymer Co., Ltd., and the like can be used.

[0142] (Cyclic olefin polymer (δ))

[0143] Hereinafter, the cyclic olefin polymer (δ) will be described, but the cyclic olefin polymer (δ) is not limited to the following embodiment.

[0144] The cyclic olefin polymer (δ) is a polymer containing a structural unit containing an alicyclic structure and not containing at least one structural unit derived from an olefin represented by the above formula (I) (excluding the cyclic olefin copolymer (α)). As the alicyclic structure in the cyclic olefin polymer (δ), a saturated cyclic hydrocarbon (cycloalkane) structure, an unsaturated cyclic hydrocarbon (cycloolefin) structure, etc. can be cited. From the viewpoints of mechanical strength, heat resistance, etc., a cycloalkane structure and a cycloolefin structure are preferred, and a cycloalkane structure is more preferred. The alicyclic structure may be located in the main chain or in the side chain. From the viewpoints of mechanical strength, heat resistance, etc., it is preferred to contain an alicyclic structure in the main chain. There is no particular limitation on the number of carbon atoms constituting the alicyclic structure. When it is generally in the range of 4 to 30, preferably 5 to 20, and more preferably 5 to 15, the characteristics of mechanical strength, heat resistance, and film formability are highly balanced.

[0145] The ratio of the structural unit containing alicyclic structure in the cyclic olefin polymer (δ) can be appropriately selected according to the purpose of use, preferably 50 mass % or more, more preferably 70 mass % or more, and particularly preferably 90 mass % or more. When the ratio of the structural unit containing alicyclic structure in the polymer containing alicyclic structure is within this range, it is preferred from the viewpoint of transparency and heat resistance of the film.

[0146] Specific examples of the cyclic olefin polymer (δ) include (1) norbornene polymers, (2) monocyclic cyclic olefin polymers, (3) cyclic conjugated diene polymers, (4) vinyl alicyclic hydrocarbon polymers, and hydrogenated products of (1) to (4). Among them, hydrogenated products of norbornene polymers, vinyl alicyclic hydrocarbon polymers, and hydrogenated products thereof are preferred from the viewpoints of heat resistance, mechanical strength, and the like.

[0147] (1) Norbornene-based polymers

[0148] Examples of the norbornene-based polymer include polymers obtained by ring-opening polymerization of a norbornene-based monomer.

[0149] Examples of the norbornene-based monomers include bicyclo[2.2.1]hept-2-ene (common name: norbornene), tricyclo[4.3.0 1,6 .1 2,5 ]Deca-3,7-diene (common name: dicyclopentadiene), 7,8-benzotricyclo[4.3.0.1 2,5 ]dec-3-ene (common name: 1,4-1,4,4a,9a-tetrahydrofluorene), tetracyclo[6.2.1.1 3,6 .0 2,7] alicyclic compounds such as dec-4-ene (common name: tetracyclododecene) and monomers having substituents (alkyl, alkylene, alkylidene, alkoxycarbonyl, etc.) on these alicyclic compounds.

[0150] Examples of polymers obtained by ring-opening polymerization include hydrogenated ring-opening polymers of norbornene-based monomers such as ZEONEX (registered trademark; manufactured by ZEON Corporation of Japan), ZEONOR (registered trademark; manufactured by ZEON Corporation of Japan), and ARTON (registered trademark; manufactured by JSR Corporation).

[0151] Among them, hydrogenated ring-opening polymers are preferred from the viewpoint of high total light transmittance, and ring-opening polymers having no polar group (ZEONEX, ZEONOR) are particularly preferred.

[0152] (2) Monocyclic cyclic olefin polymers

[0153] As the monocyclic cyclic olefin polymer, for example, an addition polymer of a monocyclic cyclic olefin monomer such as cyclohexene, cycloheptene, or cyclooctene can be used.

[0154] (3) Cyclic conjugated diene polymers

[0155] As the cyclic conjugated diene polymer, for example, a polymer obtained by 1,2- or 1,4-addition polymerization of a cyclic conjugated diene monomer such as cyclopentadiene or cyclohexadiene, and a hydrogenated product thereof, etc. can be used.

[0156] (4) Vinyl alicyclic hydrocarbon polymers

[0157] As the vinyl alicyclic hydrocarbon polymer, for example, polymers of vinyl alicyclic hydrocarbon monomers such as vinyl cyclohexene and vinyl cyclohexane and their hydrogenated products; hydrogenated products of the aromatic ring part of polymers of vinyl aromatic monomers such as styrene and α-methylstyrene, etc., random copolymers of vinyl alicyclic hydrocarbon polymers or vinyl aromatic monomers and other monomers copolymerizable with these monomers, block copolymers and their hydrogenated products, etc. can be cited. As block copolymers, diblock, triblock, or multi-blocks or tilted block copolymers thereof can be cited, without particular limitation.

[0158] (Silanized Polyolefin (ε))

[0159] The hole sidewall member may further contain a silylated polyolefin (ε). If the hole sidewall member contains a silylated polyolefin, the surface free energy of the hole sidewall member tends to decrease, which is preferred from the viewpoint of drug adsorption.

[0160] When the hole sidewall member contains the silylated polyolefin (ε), the hole sidewall member preferably contains a polyolefin-based resin, and more preferably contains a propylene-based polymer (γ) as the polyolefin-based resin.

[0161] Hereinafter, the silylated polyolefin (ε) will be described, but the silylated polyolefin (ε) is not limited to the following embodiment.

[0162] The silylated polyolefin (ε) is a reaction product of a silicon-containing compound having a structural unit represented by the following formula (1) and a vinyl-containing compound having a number average molecular weight (Mn) of 100 or more and 500,000 or less as measured by gel permeation chromatography (GPC) (excluding the case where a compound having two or more SiH groups in one molecule is used as the silicon-containing compound and a compound having an average of 2.0 or more vinyl groups per molecule is used as the vinyl-containing compound, that is, excluding the case where a silicon-containing compound having two or more SiH groups in one molecule and a vinyl-containing compound having an average of 2.0 or more vinyl groups per molecule as the vinyl-containing compound are used as the reaction product of a silicon-containing compound having two or more SiH groups in one molecule and a vinyl-containing compound having an average of 2.0 or more vinyl groups per molecule as the vinyl-containing compound, and a silylated polyolefin and its derivatives thereof are used as the reaction product of a silicon-containing compound having two or more SiH groups in one molecule and a vinyl-containing compound having an average of 2.0 or more vinyl groups per molecule as the vinyl-containing compound).

[0163] -Si(R 1 )HY 1 -···(1)

[0164] In the above formula (1), R 1 is a hydrogen atom, a halogen atom or a hydrocarbon group, Y 1 O, S or NR 30 (R 30 is a hydrogen atom or a hydrocarbon group).

[0165] <Silicon-containing compounds>

[0166] The silicon-containing compound constituting the silylated polyolefin (ε) is a hydrosilane compound having a structural unit represented by the following formula (1).

[0167] -Si(R 1 )HY 1 -···(1)

[0168] In the above formula (1), R 1 is a hydrogen atom, a halogen atom or a hydrocarbon group, Y 1 O, S or NR 30 (R 30 is a hydrogen atom or a hydrocarbon group).

[0169] Examples of the halogen atom include fluorine, chlorine, bromine and iodine.

[0170] The hydrocarbon group is preferably a hydrocarbon group having 2 to 40 carbon atoms, and more preferably a hydrocarbon group having 2 to 20 carbon atoms. Specific examples thereof include alkyl groups, alkenyl groups, and aryl groups. 1 and R 30 In the above, the hydrocarbon groups may be the same as or different from each other.

[0171] Examples of the alkyl group include straight-chain or branched alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, hexyl, 2-ethylhexyl, octyl, decyl, and octadecyl; cycloalkyl groups such as cyclopentyl, cyclohexyl, and norbornyl; and aralkyl groups such as benzyl, phenylethyl, and phenylpropyl.

[0172] Examples of the alkenyl group include vinyl, propenyl, and cyclohexenyl.

[0173] Examples of the aryl group include phenyl, tolyl, dimethylphenyl, trimethylphenyl, ethylphenyl, propylphenyl, biphenylyl, naphthyl, methylnaphthyl, anthracenyl and phenanthryl.

[0174] The above hydrocarbon groups may contain one or more heteroatoms. Specifically, there may be mentioned groups in which at least one hydrogen atom of these groups is substituted with a group containing a halogen atom, oxygen, nitrogen, silicon, phosphorus or sulfur.

[0175] In one embodiment, the silicon-containing compound has a structure represented by the following formula (2).

[0176] R 22 -(Si(R 21 )HY 21 ) m -Z-(Y 22 -Si(R 23 )H) n -R 24 ···(2)

[0177] In the above formula (2), R 21 and R 23 are each independently a hydrogen atom, a halogen atom or a hydrocarbon group,

[0178] R 22 and R 24 are each independently a halogen atom or a hydrocarbon group,

[0179] Y 21 and Y 22 Each independently is O, S or NR 30 (R 30 is a hydrogen atom or a hydrocarbon group),

[0180] m is 0 or 1,

[0181] n is 0 or 1,

[0182] When R 21 , R 23 , Y 21 and Y 22 When there are multiple groups, the groups may be the same or different.

[0183] Z is a divalent group represented by the following formula (3):

[0184] -Si(R 41 )(R 41 )-(Y 23 -Si(R 41 )(R 41 )) l -···(3)

[0185] In the above formula (3), R 41 is a hydrogen atom, a halogen atom or a hydrocarbon group, each R 41 They can be the same or different, Y 23 Each independently is O, S or NR 30 (R 30 is a hydrogen atom or a hydrocarbon group), and l is an integer from 0 to 10,000.

[0186] In the above formula (2), when m=n=0, in the above formula (3), at least one R 41 A hydrogen atom.

[0187] It should be noted that the definitions of the halogen atom and the hydrocarbon group in the above formula (2) and the above formula (3) are the same as those in the above formula (1).

[0188] It is also a typical embodiment that the hydrocarbon group in the above formulae (1), (2) and (3) is composed only of carbon atoms and hydrogen atoms.

[0189] In one embodiment, the silicon-containing compound preferably has 3 or more silicon atoms, more preferably 5 or more silicon atoms, and further preferably 10 or more silicon atoms. In addition, the silicon-containing compound preferably has 10,000 or less silicon atoms, more preferably 1,000 or less silicon atoms, particularly preferably 300 or less silicon atoms, and further preferably 50 or less silicon atoms.

[0190] In one embodiment, l in the above formula (3) is an integer of 0 to 10,000, and preferred upper and lower limits include numbers determined by the values ​​of m and n in the above formula (2) and the above preferred number of silicon atoms.

[0191] In one embodiment, it is preferable to use a silicon-containing compound in which m=n=1 in the above formula (2), that is, a compound having SiH groups at both ends.

[0192] In one embodiment, it is preferable to use a silicon-containing compound in which m=1 and n=0 in the above formula (2), that is, a compound having a SiH group at one end.

[0193] Particularly preferred silicon-containing compounds include those in the above formula (2) and formula (3) wherein m=n=1 and R 21 , R23 and R 41 A compound in which all of the hydrocarbon groups are hydrocarbon groups. The hydrocarbon groups may be the same or different.

[0194] As particularly preferred other silicon-containing compounds, there are those wherein m=1, n=0 and R 21 and R 41 A compound in which all of the hydrocarbon groups are hydrocarbon groups. The hydrocarbon groups may be the same or different.

[0195] Specific examples of the silicon-containing compound used in the present embodiment are shown below. Examples of the silicon-containing compound in the present embodiment include compounds having one SiH group.

[0196] Examples of silicon-containing compounds having one SiH group include compounds represented by the following formula (2a), compounds in which a part or all of the methyl groups in formula (2a) are substituted by hydrocarbon groups which may contain one or more hetero atoms, such as ethyl, propyl, phenyl, trifluoropropyl, etc.

[0197] HSi(CH3)2O-(-Si(CH 3 )2-O-) d -Si(CH3)3···(2a)

[0198] (In the above formula (2a), d is an integer greater than or equal to 1, and the upper limit is, for example, 1000, preferably 300, and more preferably 50.)

[0199] More specifically, examples of such a compound include the following compounds, but the compound is not limited to these compounds.

[0200] C4H9-((CH3)2SiO)9-(CH3)2SiH

[0201] C4H9-((CH3)2SiO) 65 -(CH3)2SiH

[0202] As another example of a silicon-containing compound having one SiH group, there can be mentioned, for example, a dimethylsiloxane-methylhydrogensiloxane copolymer represented by the following formula (2b), a compound in which a part or all of the methyl groups in the following formula (2b) are substituted by a hydrocarbon group which may contain one or more hetero atoms, such as an ethyl group, a propyl group, a phenyl group, or a trifluoropropyl group.

[0203] Si(CH3)3O-(-Si(CH3)2-O-) e -(-SiH(CH3)-O-)-Si(CH3)3···(2b)

[0204] (In the above formula (2b), e is an integer greater than or equal to 0, and the upper limit is, for example, 1000, preferably 300, and more preferably 50.)

[0205] In addition, the arrangement order of -Si(CH3)2-O- units and -SiH(CH3)-O- units is not particularly limited and can be block, disordered, or statistically random.

[0206] More specifically, examples of such a compound include the following compounds, but the compound is not limited thereto.

[0207] Si(CH3)3O-SiH(CH3)-O-Si(CH3)3

[0208] Examples of the silicon-containing compound of the present embodiment include compounds having two or more SiH groups.

[0209] Examples of silicon-containing compounds having two or more SiH groups include methyl hydrogen polysiloxane represented by the following formula (2c), compounds in which a part or all of the methyl groups in the following formula (2c) are substituted by hydrocarbon groups that may contain one or more hetero atoms, such as ethyl, propyl, phenyl, trifluoropropyl, etc.

[0210] (CH3)3SiO-(-SiH(CH3)-O-) f -Si(CH3)3···(2c)

[0211] (In the above formula (2c), f is an integer greater than or equal to 2, and the upper limit is, for example, 1000, preferably 300, and more preferably 50.)

[0212] As another example of a silicon-containing compound having two or more SiH groups, for example, there can be mentioned a dimethylsiloxane-methylhydrogensiloxane copolymer represented by the following formula (2d), a compound in which a part or all of the methyl groups in the following formula (2d) are substituted by a hydrocarbon group which may contain one or more hetero atoms, such as an ethyl group, a propyl group, a phenyl group, or a trifluoropropyl group.

[0213] (CH3)3SiO-(-Si(CH3)2-O-) g -(-SiH(CH3)-O-) h -Si(CH3)3···(2d)

[0214] (In the above formula (2d), g is an integer greater than or equal to 1, and h is an integer greater than or equal to 2. The upper limit of the total of g and h is, for example, 1000, preferably 300, and more preferably 50.)

[0215] In addition, in the above formula (2d), the order of arrangement of -Si(CH3)2-O- units and -SiH(CH3)-O- units is not particularly limited and can be block, disordered, or statistically random.

[0216] More specifically, examples of such a compound include the following compounds, but the compound is not limited thereto.

[0217] [Chemistry 9]

[0218]

[0219] As another example of a silicon-containing compound having two or more SiH groups, for example, there can be mentioned methyl hydrogen polysiloxane represented by the following formula (2e), a compound in which a part or all of the methyl groups in the following formula (2e) are substituted by a hydrocarbon group which may contain one or more hetero atoms, such as an ethyl group, a propyl group, a phenyl group, or a trifluoropropyl group.

[0220] HSi(CH3)2O-(-Si(CH3)2-O-) i -Si(CH3)2H···(2e)

[0221] (In the above formula (2e), i is an integer greater than or equal to 1, and the upper limit is, for example, 1000, preferably 300, and more preferably 50.)

[0222] More specifically, examples of such a compound include the following compounds, but the compound is not limited to these compounds.

[0223] HSi(CH3)2O-(-Si(CH3)2-O-)5-Si(CH3)2H

[0224] HSi(CH3)2O-(-Si(CH3)2-O-)8-Si(CH3)2H

[0225] HSi(CH3)2O-(-Si(CH3)2-O-) 18 -Si(CH3)2H

[0226] HSi(CH3)2O-(-Si(CH3)2-O-) 80 -Si(CH3)2H

[0227] HSi(CH3)2O-(-Si(CH3)2-O-) 230 -Si(CH3)2H

[0228] As another example of a silicon-containing compound having two or more SiH groups, for example, there can be mentioned methyl hydrogen polysiloxane represented by the following formula (2f), a compound in which a part or all of the methyl groups in the following formula (2f) are substituted by a hydrocarbon group which may contain one or more hetero atoms, such as an ethyl group, a propyl group, a phenyl group, or a trifluoropropyl group, etc.

[0229] HSi(CH3)2O-(-SiH(CH3)-O-) j -Si(CH3)2H···(2f)

[0230] (In the above formula (2f), j is an integer greater than or equal to 1, and the upper limit is, for example, 1000, preferably 300, and more preferably 50.)

[0231] As another example of a silicon-containing compound having two or more SiH groups, there can be cited, for example, a dimethylsiloxane-methylhydrogensiloxane copolymer represented by the following formula (2g), a compound in which a part or all of the methyl groups in the following formula (2g) are substituted by a hydrocarbon group which may contain one or more hetero atoms, such as an ethyl group, a propyl group, a phenyl group, or a trifluoropropyl group.

[0232] HSi(CH3)2O-(-Si(CH3)2-O-) k -(-SiH(CH3)-O-) l -Si(CH3)2H···(2g)

[0233] (In the above formula (2g), k and l are each an integer greater than or equal to 1, and the upper limit of the total of k and l is, for example, 1000, preferably 300, and more preferably 50.)

[0234] In addition, the order of arrangement of -Si(CH3)2-O- units and -SiH(CH3)-O- units is not particularly limited and may be block-like, disordered, or statistically random.

[0235] The number average molecular weight (Mn) of the vinyl compound measured by gel permeation chromatography (GPC) is generally 100 or more and 500,000 or less, more preferably 100 or more and 100,000 or less. When the number average molecular weight is above the above lower limit, the silylated polyolefin obtained can be further suppressed from seeping out from the material constituting the hole side wall component. When it is below the above upper limit, the dispersibility of the silylated polyolefin in the material constituting the hole side wall component is improved, and the deviation of the surface free energy of the plate can be reduced. It should be noted that in the present embodiment, as described later, the number average molecular weight (Mn), the weight average molecular weight (Mw) and Mw / Mn are values ​​converted to polyethylene.

[0236] <Vinyl Compounds>

[0237] The vinyl-containing compound constituting the silylated polyolefin (ε) is usually obtained by polymerizing or copolymerizing one or more olefins selected from olefins having 2 to 50 carbon atoms.

[0238] Specific examples of the olefins having 2 to 50 carbon atoms include α-olefins such as ethylene, propylene, 1-butene, 1-pentene, 3-methyl-1-butene, 1-hexene, 4-methyl-1-pentene, 3-methyl-1-pentene, 3,4-dimethyl-1-pentene, 4-methyl-1-hexene, 3-ethyl-1-pentene, 3-ethyl-4-methyl-1-pentene, 3,4-dimethyl-1-hexene, 4-methyl-1-heptene, 3,4-dimethyl-1-heptene, 1-octene, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, 1-eicosene, and vinylcyclohexane; olefins having an internal double bond such as cis-2-butene and trans-2-butene; isobutylene, 2-butylene, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, 1-eicosene, and vinylcyclohexane; olefins having an internal double bond such as cis-2-butene and trans-2-butene; and isobutylene, 2-butylene, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, 1-eicosene, and vinylcyclohexane. 2,4-Dimethyl-1-pentene, 2,4-dimethyl-1-hexene, 2,4,4-trimethyl-1-pentene, 2,4-dimethyl-1-heptene, 2-methyl-1-butene, 2-methyl-1-hexene, 2-methyl-1-heptene, 2-methyl-1-octene, 2,3-dimethyl-1-butene, 2,3-dimethyl-1-pentene, 2,3-dimethyl-1-hexene, 2,3-dimethyl-1-octene, 2,3,3-trimethyl-1-butene, 2,3,3-trimethyl-1-pentene, 2,3,3-trimethyl-1-hexene, 2,3,3-trimethyl-1-octene, 2,3,4-trimethyl-1-pentene, 2,3,4-trimethyl-1-hexene, 2 , 3,4-trimethyl-1-octene, 2,4,4-trimethyl-1-hexene, 2,4,4-trimethyl-1-octene, 2-methyl-3-cyclohexyl-1-propene, vinylidene cyclopentane, vinylidene cyclohexane, vinylidene cyclooctane, 2-methylvinylidene cyclopentane, 3-methylvinylidene cyclopentane, 4-methylvinylidene cyclopentane and other vinylidene compounds; styrene, o-methylstyrene, m-methylstyrene, p-methylstyrene, 2,4-dimethylstyrene, o-ethylstyrene, m-ethylstyrene, p-ethylstyrene and other aromatic vinyl compounds; α-methylstyrene, α-ethylstyrene, 2-methyl-3-phenylpropene and other aromatic vinylidene compounds; methyl methacrylate, ethyl methacrylate Ester, n-propyl methacrylate, isopropyl methacrylate, n-butyl methacrylate, isobutyl methacrylate, tert-butyl methacrylate, 2-cyanopropylene, 2-aminopropylene, 2-hydroxymethylpropylene, 2-fluoropropylene, 2-chloropropylene and other functional group substituted vinylidene compounds; cyclobutene, cyclopentene, 1-methyl-1-cyclopentene, 3-methyl-1-cyclopentene, 2-methyl-1-cyclopentene, cyclohexene, 1-methyl-1-cyclohexene, 3-methyl-1-cyclohexene, 2-methyl-1-cyclohexene, cycloheptene, cyclooctene, norbornene, 5-methyl-2-norbornene, tetracyclododecene, 5,6-dihydrodicyclopentadiene, 3a,4,5,6,7,7a-hexahydro-H-indene, tricyclo[6.2.1.0 2,7] aliphatic cyclic olefins containing internal double bonds such as undecene, cyclopentadiene, and dicyclopentadiene; cyclic olefins containing aromatic rings such as cyclopent-2-enylbenzene, cyclopent-3-enylbenzene, cyclohex-2-enylbenzene, cyclohex-3-enylbenzene, indene, 1,2-dihydronaphthalene, 1,4-dihydronaphthalene, 1,4-bridged methyl-1,4,4a,9a-tetrahydrofluorene; butadiene, isoprene, 4-methyl-1,3-pentadiene, 4-methyl-1,4-pentadiene, 1,3-pentadiene, 1,4- Cyclic polyenes having two or more double bonds, such as pentadiene, 1,5-hexadiene, 1,4-hexadiene, 1,3-hexadiene, 1,3-octadiene, 1,4-octadiene, 1,5-octadiene, 1,6-octadiene, 1,7-octadiene, ethylidene norbornene, vinyl norbornene, dicyclopentadiene, 7-methyl-1,6-octadiene, 4-ethylidene-8-methyl-1,7-nonadiene, and 5,9-dimethyl-1,4,8-decatriene, and chain polyenes having two or more double bonds.

[0239] In addition, the olefin having 2 to 50 carbon atoms may have a functional group containing an atom such as oxygen, nitrogen, or sulfur. Examples thereof include unsaturated carboxylic acids such as acrylic acid, fumaric acid, itaconic acid, and bicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid, and unsaturated carboxylic acid metal salts such as sodium salts, potassium salts, lithium salts, zinc salts, magnesium salts, and calcium salts thereof; unsaturated carboxylic acid anhydrides such as maleic anhydride, itaconic anhydride, and bicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid anhydride; unsaturated carboxylic acid esters such as methyl acrylate, ethyl acrylate, n-propyl acrylate, isopropyl acrylate, n-butyl acrylate, isobutyl acrylate, tert-butyl acrylate, and 2-ethylhexyl acrylate; and vinyl esters such as vinyl acetate, vinyl propionate, vinyl caproate, vinyl caprate, vinyl laurate, vinyl stearate, and vinyl trifluoroacetate. unsaturated glycidyl esters such as glycidyl acrylate, glycidyl methacrylate, monoglycidyl itaconate; halogenated olefins such as vinyl chloride, vinyl fluoride, fluorinated allyl; unsaturated cyano compounds such as acrylonitrile and 2-cyano-bicyclo[2.2.1]hept-5-ene; unsaturated ether compounds such as methyl vinyl ether and ethyl vinyl ether; unsaturated amides such as acrylamide, methacrylamide, N,N-dimethylacrylamide; styrene derivatives containing functional groups such as methoxystyrene, ethoxystyrene, vinyl benzoic acid, methyl vinyl benzoate, vinyl benzyl acetate, hydroxystyrene, o-chlorostyrene, p-chlorostyrene, divinylbenzene, etc.; N-vinylpyrrolidone, etc.

[0240] In a preferred embodiment, the vinyl group-containing compound has a structure represented by the following formula (4) and has a number average molecular weight of 100 or more and 500,000 or less.

[0241] A-CH=CH2···(4)

[0242] Here, in the above formula (4), A is a polymer chain containing one or more structural units derived from an α-olefin having 2 to 50 carbon atoms.

[0243] In the above formula (4), the A part of the vinyl group-containing compound is preferably an ethylene polymer chain, a propylene polymer chain or a copolymer chain of two or more olefins selected from the group consisting of α-olefins having 2 to 50 carbon atoms. The α-olefin is preferably an α-olefin having 2 to 20 carbon atoms.

[0244] In a preferred embodiment, A of the vinyl-containing compound represented by the above formula (4) is a polymer chain consisting only of α-olefins having 2 to 50 carbon atoms. More preferably, A of the vinyl-containing compound is a polymer chain consisting only of α-olefins having 2 to 20 carbon atoms. More preferably, A of the vinyl-containing compound is an ethylene homopolymer chain, a propylene homopolymer chain, or a copolymer chain of ethylene and an α-olefin having 3 to 20 carbon atoms.

[0245] The vinyl-containing compound represented by the above formula (4) is preferably an ethylene-α-olefin copolymer in which the structural unit derived from ethylene is in the range of 81 to 100 mol%, and the structural unit derived from α-olefin having 3 to 20 mol% of carbon atoms is in the range of 0 to 19 mol%. More preferably, it is an ethylene-α-olefin copolymer in which the structural unit derived from ethylene is in the range of 90 to 100 mol%, and the structural unit derived from α-olefin having 3 to 20 mol% of carbon atoms is in the range of 0 to 10 mol%. Among them, the total of the structural unit derived from ethylene and the structural unit derived from α-olefin having 3 to 20 carbon atoms is 100 mol%. It is particularly preferred that the structural unit derived from ethylene is 100 mol%.

[0246] The molecular weight distribution (ratio of weight average molecular weight to number average molecular weight, Mw / Mn) of the vinyl group-containing compound represented by the formula (4) as measured by gel permeation chromatography (GPC) is preferably in the range of 1.1 to 3.0.

[0247] The number average molecular weight (Mn) of the vinyl-containing compound represented by the above formula (4) is preferably in the range of 100 to 500,000, more preferably 100 to 100,000, further preferably 500 to 50,000, and further preferably 700 to 10,000.

[0248] The melting point of the vinyl group-containing compound represented by the above formula (4) is preferably 70° C. or higher and 130° C. or lower.

[0249] It is more preferred that the vinyl group of the vinyl-containing compound represented by the above formula (4) is present at the terminal of the main chain, and it is more preferred that the vinyl group is present only at the terminal of the main chain.

[0250] When the vinyl-containing compound represented by the above formula (4) contains vinyl groups only at the ends of the main chain, 1 The terminal unsaturation rate calculated by H-NMR is preferably 60 mol% or more and 100 mol% or less. One of the further preferred embodiments is to 1 The terminal unsaturation ratio calculated by H-NMR is 80 mol% to 99.5 mol%, more preferably 90 mol% to 99 mol%.

[0251] As the vinyl-containing compound represented by the formula (4), for example, those described in JP-A-2017-155159, in particular, in paragraphs

[0117] to

[0156] of the publication can be adopted.

[0252] The silylated polyolefin used in the present embodiment may be one produced by any method, and for example, examples described in JP-A-2017-155159, particularly paragraphs

[0159] to

[0189] of the publication may be used.

[0253] It is estimated that the silylated polyolefin (ε) has structures represented by, for example, formulae (5) to (9). Of course, the combination of the silicon-containing compound and the vinyl-containing compound is not limited to the above examples.

[0254] [Chemistry 10]

[0255]

[0256] [Chemistry 11]

[0257]

[0258] [Chemistry 12]

[0259]

[0260] [Chemistry 13]

[0261]

[0262] [Chemistry 14]

[0263]

[0264] As the silylated polyolefin (ε), a commercially available silylated polyolefin can be used, and as a commercially available silylated polyolefin, the Exfola series manufactured by Mitsui Chemicals, Inc. can be used.

[0265] The content of the silylated polyolefin (ε) in the material constituting the hole side wall member is preferably 1 to 20% by mass, more preferably 5 to 15% by mass, based on 100% by mass of the material constituting the hole side wall member.

[0266] (Glass)

[0267] As the bottom surface member, glass can be used, and examples thereof include quartz glass, borosilicate glass, phosphate glass, chemically strengthened glass, etc. The glass constituting the bottom surface member may be one type, or two or more types.

[0268] A commercially available item can be used as the glass, and as the commercially available glass, for example, NEO Cover Glass manufactured by Matsunami Glass Industries, Ltd. can be used.

[0269] From the viewpoint of drug adsorption, one of the preferred embodiments of the orifice plate (X) is that the orifice sidewall member comprises a cyclic olefin copolymer (α) or a propylene polymer (γ), and the bottom member comprises a 4-methyl-1-pentene polymer (β). In addition, it is one of the preferred embodiments that the orifice sidewall member comprises a 4-methyl-1-pentene polymer (β), in which case it is more preferred that the bottom member comprises a cyclic olefin copolymer (α), a propylene polymer (γ), a cyclic olefin polymer (δ) or borosilicate glass. One of the more preferred embodiments of the orifice plate (X) is that the orifice sidewall member comprises a propylene polymer (γ) and a silanized polyolefin (ε), and the bottom member comprises a 4-methyl-1-pentene polymer (β).

[0270] <Methods for culturing cells, tissues or organs>

[0271] One embodiment of the present invention is a method for culturing cells, tissues, or organs, comprising the step of culturing cells, tissues, or organs using a well plate (X).

[0272] The method for culturing cells and the like is not particularly limited and may be carried out according to a known protocol, or a commercially available culture medium or culture kit may be used.

[0273] Cultivation is usually performed in a culture medium placed in the wells of a well plate (X). Cultivation in a culture medium means culturing with at least a portion of the cells or the like immersed in the culture medium, but the entire cells or the like may not be immersed in the culture medium.

[0274] The culture medium is not particularly limited as long as it is a culture medium in which cells etc. can proliferate, and may be appropriately selected depending on the type of cells etc. to be used.

[0275] The amount of the medium used in the culture and the frequency of medium replacement are not particularly limited. The culture temperature is not particularly limited, but is usually about 25°C to 40°C.

[0276] The culture time is not particularly limited, and the cells may be cultured until the cells and the like are sufficiently attached to the culture container and a desired confluence is achieved.

[0277] Example

[0278] Hereinafter, the present disclosure will be described in more detail with reference to Examples and Comparative Examples, but the present disclosure is not limited in any way.

[0279] [Manufacturing Example 1] Manufacture of orifice plate frame (A-1)

[0280] As the propylene polymer (γ), NOVATEC BC03C from Japan Polypropylene Co., Ltd. was used to form a 24-well plate frame with open bottom surfaces by injection molding. The surface free energy of the molded well plate frame (A-1) was 32.2 mN / m. The method for measuring the surface free energy is described below.

[0281] [Manufacturing Example 2] Manufacture of orifice plate frame (A-2)

[0282] A 24-well plate frame was molded in the same manner as in Production Example 1 except that APEL 6015T manufactured by Mitsui Chemicals, Inc. was used as the cyclic olefin copolymer (α). The surface free energy of the molded well plate frame (A-2) was 29.0 mN / m.

[0283] [Manufacturing Example 3] Manufacture of orifice plate frame (A-3)

[0284] A 24-well plate frame was molded in the same manner as in Production Example 1 except that 90 parts by mass of Prime Polymer polypropylene F107 as the propylene polymer (γ) was dry-blended with 10 parts by mass of the silylated polyolefin obtained in Synthesis Example 6 of Japanese Patent No. 5575929 as the silylated polyolefin (ε) instead of the propylene polymer (γ). The surface free energy of the molded well plate frame (A-3) was 24.7 mN / m.

[0285] [Manufacturing Example 4] Manufacture of orifice plate frame (A-4)

[0286] As the 4-methyl-1-pentene polymer (β), TPX (total light transmittance 92%, glass transition temperature 40° C., weight average molecular weight (Mw) 42.8×10 4 ), except for this, a 24-well plate frame was formed in the same manner as in Manufacturing Example 1. The surface free energy of the formed well plate frame (A-4) was 24.0 mN / m. The determination methods of total light transmittance, glass transition temperature and weight average molecular weight (Mw) are described below.

[0287] [Manufacturing Example 5] Manufacture of bottom surface members (B-1) and (B-2)

[0288] As the 4-methyl-1-pentene polymer (β), TPX (total light transmittance 93.7%, glass transition temperature 40°C, weight average molecular weight (Mw) 42.8×10 4 ), put it into an extruder with a T-die having a full-thread screw for extruding the substrate layer, set the extrusion temperature to 270°C, set the roller temperature to 60°C, and change the roller rotation speed to perform extrusion molding, thereby obtaining two films with different thicknesses. The film with a thickness of 50 μm is used as the bottom surface member (B-1), and the film with a thickness of 400 μm is used as the bottom surface member (B-2).

[0289] The content of the 4-methyl-1-pentene polymer (β) was 99.5% by mass in 100% by mass of the bottom member (B-1) or (B-2). In addition, when the total amount of repeating structural units of the 4-methyl-1-pentene polymer (β) was 100% by mole, the content of the structural unit derived from 4-methyl-1-pentene was 98.4% by mole.

[0290] [Manufacturing Example 6] Manufacture of bottom member (B-5)

[0291] A pellet of a norbornene polymer (a hydrogenated product of a ring-opening polymer of a norbornene monomer, corresponding to a cyclic olefin polymer (δ), ZEONOR1420, manufactured by ZEON Corporation of Japan; glass transition temperature Tg: 136° C.) was dried at 100° C. for 4 hours using a hot air dryer through which air was circulated. The pellet was then extruded at 260° C. using a single screw extruder equipped with a polymer filter (filtration accuracy 30 μm) and a T-die, and the extruded sheet of norbornene polymer was passed through a cooling drum for cooling to obtain a cyclic olefin polymer (δ) film having a thickness of 50 μm as a bottom member (B-5).

[0292] [Manufacturing Example 7] Manufacture of culture container

[0293] A polypropylene film having a thickness of 100 μm (corresponding to the propylene-based polymer (γ), CTP1147 manufactured by Toyobo Co., Ltd.) was used as the bottom member (B-3).

[0294] A cyclic olefin copolymer film having a thickness of 25 μm (corresponding to the cyclic olefin copolymer (α), COXEC TCS-1 manufactured by Kurabo Co., Ltd.) was used as the bottom member (B-4).

[0295] NEO Cover glass (manufactured by Matsunami Glass Industries, Ltd., product code C024601) was used as the bottom surface member (B-6).

[0296] For bottom surface members (B-1) to (B-6), a normal pressure plasma surface treatment apparatus (manufactured by Sekisui Chemical Co., Ltd.) was used to perform plasma treatment (treatment speed 2 m / min, output 4.5 kW, 2 reciprocating times) by filling the chamber with a nitrogen flow.

[0297] The bottom surface members (B-1) to (B-6) after plasma treatment are adhered to the bottom surface of the well plate frame (A-1) to (A-4) with the aid of a medical adhesive (manufactured by 3M Company) to obtain a well plate. The obtained well plate is packaged in a γ-ray resistant bag, irradiated with 10 kGy of γ-rays for sterilization, and then a 0.5 mg / mL collagen solution is added to the wells of the well plate at 0.5 mL / well, and the excess collagen solution is removed. After standing at room temperature for 30 to 60 minutes, wash with Dulbecco PBS (-), dry overnight at room temperature, and prepare culture containers 1 to 8. The combination of the well plate frames (A-1) to (A-4) and the bottom surface members (B-1) to (B-6) used in the culture containers 1 to 8 is shown in Table 1 described later. The culture containers 1 to 8 are respectively referred to as Examples 1 to 8.

[0298] (Evaluation of cell adhesion)

[0299] The cell suspension of rat primary frozen hepatocytes was added to culture vessels 1 to 8 at a volume of 0.5 mL / well to a cell density of 1×10 5 cells / cm 2 The cells were sown in a manner of , incubated at 37°C, 5% CO2, and the state after 1 day and 7 days of culture was observed under a microscope and evaluated according to the following criteria. The results are shown in Table 1.

[0300] A: Hepatocytes were observed to be adhered to the culture surface and spread out.

[0301] B: The hepatocytes were observed to be adhered to the culture surface but rounded and not stretched, or partially detached. Or the hepatocytes were not adhered to the culture surface.

[0302] (Evaluation of drug adsorption)

[0303] For any three wells of the culture containers 1 to 8, 0.5 mL of the evaluation drug solution was added to each well, and the drug solution was recovered after standing at 23°C for 2 days. The concentration of the recovered evaluation drug solution 1 was measured by LC / MS, and the ratio (%) to the concentration of the evaluation drug solution 1 before adding to the culture containers 1 to 8 was calculated, and the average value in the three wells was obtained as the drug residual rate. The results are shown in Table 1. The higher the drug residual rate, the lower the drug adsorption property of the plate.

[0304] The evaluation drug solution 1 was changed to evaluation drug solution 2, and the operation was carried out in the same way except that 3 wells were randomly selected from 21 wells that had not been used in the evaluation of evaluation drug solution 1, and the drug residue rate was calculated. The results are shown in Table 1.

[0305] Evaluation drug solution 1: Dimethyl sulfoxide (hereinafter referred to as DMSO) solution of cyclosporine A (concentration 10 μmol / L)

[0306] Evaluation drug solution 2: DMSO solution of isoproterenol hydrochloride (concentration 10 μmol / L)

[0307] <LC / MS conditions>

[0308] · Instrument: Acquity UPLC I-class system / TQ-S micro (water)

[0309] · Ionization method: Electrospray ionization (ESI), positive and negative ion detection

[0310] · Detection: Selected reaction monitoring (SRM)

[0311] 1. Polarity:

[0312] Positive: Cyclosporine A, isoproterenol hydrochloride

[0313] 2. Precursor ion:

[0314] Cyclosporine A: m / z1203

[0315] Isoproterenol hydrochloride: m / z212

[0316] 3. Product ion:

[0317] Cyclosporine A: m / z156

[0318] Isoproterenol hydrochloride: m / z152

[0319] (Measurement of surface free energy)

[0320] Using an A-XE type (manufactured by Kyowa Interface Chemical Co., Ltd.) contact angle meter, water, diiodomethane, and 1-bromonaphthalene were used as standard test solutions 1 to 3 to measure the contact angle of the well plate frame (test object). The surface free energy of the test object was calculated using the known surface free energy of the standard test solution and the measured value of the contact angle of the standard test solution on the test object by a calculation method based on the Kitazaki-Hata theory described later for calculating the surface free energy. The calculation was performed using the application software attached to the contact angle meter, namely the multifunctional comprehensive analysis software FAMAS (manufactured by Kyowa Interface Chemical Co., Ltd.). In addition, the numerical values ​​recorded in Table 1 record the average value of the results of three contact angle measurements performed on one test object in the same test.

[0321] The following describes Kitazaki-Hata's theory in more detail.

[0322] When the surface free energy of the orifice frame to be calculated is set to A, and the components of the surface free energy A are set to the component attributable to the dispersion force (Ad), the component attributable to the orientation (polar) force (Ap), and the component attributable to the hydrogen bonding force (A1h), A is as shown in formula (10). The intermolecular force caused by the inductive force is extremely small and is therefore ignored in the calculation.

[0323] [Number 1]

[0324] A=A d +A p +A h Formula (10)

[0325] In addition, the surface free energy of the standard test solution 1 whose values ​​of each component are known is referred to as B1, and the component of the surface free energy B1 is referred to as the component attributable to the dispersion force (B 1d ), the component attributable to the orientation (polarity) force (B 1p ) and the component attributable to hydrogen bonding (B 1h ), B1 is as shown in formula (11).

[0326] [Number 2]

[0327] B1=B 1d +B 1p +B 1h Formula (11)

[0328] Similarly, when the surface free energy of the standard test liquid 2 whose values ​​of each component are known is set to B2 and the surface free energy of the liquid 3 whose values ​​of each component are known is set to B3, B2 and B3 are expressed as equations (12) and (13), respectively.

[0329] [Number 3]

[0330] B2=B 2d +B 2p +B2h Formula (12)

[0331] [Number 4]

[0332] B3=B 3d +B 3p +B 3h Formula (13)

[0333] Furthermore, when the contact angle measured using the orifice plate frame and standard test solution 1 is set to θ1, the contact angle between the orifice plate frame and standard test solution 2 is set to θ2, and the contact angle between the orifice plate frame and standard test solution 3 is set to θ3, for the values ​​of the contact angles and components of the surface free energy of the orifice plate frame and standard test solution 1, standard test solution 2, and standard test solution 3, the relationships of equations (14), (15), and (16) hold.

[0334] [Number 5]

[0335]

[0336] [Number 6]

[0337]

[0338] [Number 7]

[0339]

[0340] By solving the three-variable linear equations composed of the above equations (14), (15) and (16), A d , A p and A h , and then calculate the surface free energy A of the orifice frame according to formula (10).

[0341] (Measurement of glass transition temperature of bottom member)

[0342] The glass transition temperatures (Tg) of the bottom surface members of the culture containers 1 to 7 were measured using a differential scanning calorimeter in accordance with Japanese Industrial Standard JIS-K7121:1987 (Method for measuring transition temperatures of plastics).

[0343] The glass transition temperature (Tg) of the borosilicate glass used as the bottom member of the culture container 8 was measured using a differential thermal dilatometer (TMA) in accordance with Japanese Industrial Standard JIS-R3103-3 (transition temperature measurement method based on thermal expansion method).

[0344] (Measurement of glass transition temperature of hole side wall member)

[0345] The glass transition temperature (Tg) of the hole side wall members of the culture containers 1 to 7 is obtained by performing solid viscoelastic temperature dispersion measurement under the following conditions, and the peak value of the loss tangent (tanδ) obtained is set as the glass transition temperature (Tg) of the hole side wall member.

[0346] (Measurement conditions)

[0347] Measuring device: DVA-225 manufactured by IT Measurement and Control Co., Ltd.

[0348] Deformation Mode: Stretch

[0349] Temperature range: -30℃~200℃

[0350] Heating rate: 3℃ / min

[0351] Frequency: 1Hz

[0352] Strain: 0.05%

[0353] Atmosphere: Atmosphere

[0354] (Measurement of total light transmittance)

[0355] About the measurement sample, the total light transmittance was measured using a haze meter (NDH2000, manufactured by Nippon Denshoku Industries Co., Ltd.) in accordance with JIS K7361-1.

[0356] [Measurement of water contact angle]

[0357] The water contact angle is measured in accordance with Japanese Industrial Standard JIS-R3257 (Wettability Test Method for Substrate Glass Surface). Under constant temperature and humidity conditions of 25±5°C and 50±10%, a water drop of 4 μL or less is dripped onto the surface of the sample so that the shape of the water drop can be regarded as a sphere, and the angle of the contact interface between the sample and the water drop is measured by the static drop method within 1 minute after the water drop contacts the sample surface.

[0358] [Comparative Example 1]

[0359] A commercially available 24-well TCPS (tissue culture polystyrene) culture container (manufactured by Corning, model NCO3524, bottom member and well plate frame made of surface hydrophilized polystyrene) with a bottom thickness of 1000 μm was used as culture container 9. Evaluation was carried out in the same manner as in Examples 1 to 8 except that the culture container was different. The results are shown in Table 1.

[0360] [Comparative Example 2]

[0361] As a high oxygen permeable container, a commercially available 24-well culture container (product name G-plate, manufactured by VECELL, model number V24WGPB-10, bottom member made of PDMS (polydimethylsiloxane), well plate frame made of surface hydrophilized polystyrene) with a bottom member thickness of 350 μm was used as the culture container 10. Except for the difference in the culture container, the evaluation was carried out in the same manner as in Examples 1 to 8. The results are shown in Table 1.

[0362] [Comparative Example 3]

[0363] For the bottom member (B-1), a normal pressure plasma surface treatment device (manufactured by Sekisui Chemical Co., Ltd.) was used to fill the chamber with a nitrogen flow and perform plasma treatment (treatment speed 2m / min, output power 4.5kW, reciprocation 2 times). By peeling off the bottom member of the culture container 10, a surface hydrophilized polystyrene 24-well plate frame (surface free energy of 43.0mN / m) with the bottom opening of the hole was prepared, and the bottom member (B-1) after plasma treatment was closely attached to the bottom surface with the help of a medical adhesive (manufactured by 3M Company) to obtain a well plate. The obtained well plate was packaged in a γ-ray resistant bag, irradiated with 10kGy of γ-rays for sterilization, and then 0.5mg / mL of collagen solution was added to the wells of the well plate at 0.5mL / well, and the excess collagen solution was removed. After standing at room temperature for 30 to 60 minutes, it was washed with Dulbecco PBS (-) and dried overnight at room temperature to obtain the well plate of Comparative Example 3 as the culture container 11. The obtained well plates were evaluated in the same manner as in Examples 1 to 8 except that the culture containers were different. The results are shown in Table 1.

[0364] [Comparative Example 4]

[0365] A well plate was prepared in the same manner as in Comparative Example 3, except that the bottom member (B-3) was used instead of the bottom member (B-1), as the culture container 12. Except for the difference in the culture container, the evaluation was carried out in the same manner as in Examples 1 to 8. The results are shown in Table 1.

[0366] [Table 1]

[0367]

[0368] In the culture containers 1 to 8, the adhesion of cells and the like is excellent and the drug adsorption is low. In particular, in the culture container 4 whose hole side wall member comprises a propylene polymer (γ) and a silylated polyolefin (ε) and whose bottom surface member comprises a 4-methyl-1-pentene polymer (β), the culture container 5 whose hole side wall member comprises a 4-methyl-1-pentene polymer (β) and whose bottom surface member comprises a propylene polymer (γ), the culture container 6 whose hole side wall member comprises a 4-methyl-1-pentene polymer (β) and whose bottom surface member comprises a cyclic olefin copolymer (α), the culture container 7 whose hole side wall member comprises a 4-methyl-1-pentene polymer (β) and whose bottom surface member comprises a cyclic olefin polymer (δ), and the culture container 8 whose hole side wall member comprises a 4-methyl-1-pentene polymer (β) and whose bottom surface member comprises a borosilicate glass, the adhesion of cells and the like is excellent and the drug adsorption is very low. On the other hand, although the culture vessels 9 to 12 have excellent adhesion to cells and the like, they have high drug adsorption properties and are not satisfactory for use as culture vessels.

Claims

1. A hole plate having a bottom surface member and a hole side wall member, At least one hole is formed by the bottom surface member and the hole side wall member being joined together, The bottom surface member and the hole side wall member are made of different materials, The surface free energy of the hole side wall member is 18.0 to 34.0 mN / m.

2. The orifice plate according to claim 1, wherein The hole sidewall member includes a thermoplastic resin.

3. The orifice plate according to claim 2, wherein: The thermoplastic resin is a polyolefin-based resin.

4. The orifice plate according to claim 3, wherein: The polyolefin resin is at least one selected from a cyclic olefin copolymer (α), a 4-methyl-1-pentene polymer (β), a propylene polymer (γ) and a cyclic olefin polymer (δ), wherein the cyclic olefin polymer (δ) does not include the cyclic olefin copolymer (α).

5. The orifice plate according to claim 4, wherein: The hole sidewall member further comprises a silanized polyolefin (ε).

6. The orifice plate according to claim 1 or 2, wherein: The bottom surface member has a glass transition temperature of -40°C or higher as measured in accordance with JIS-K7121:1987.

7. The orifice plate according to claim 1 or 2, wherein: The total light transmittance of the bottom member measured in accordance with JIS-K-7361-1 is 80.0% or more.

8. The orifice plate according to claim 1 or 2, wherein: The bottom surface member has a thickness of 20 to 500 μm.

9. The orifice plate according to claim 1 or 2, wherein: The culture surface of the bottom member is coated with at least one coating agent selected from natural polymer materials, synthetic polymer materials and inorganic materials.

10. The orifice plate according to claim 1 or 2, wherein: The water contact angle of the culture surface of the bottom member is 50 to 100°.

11. The orifice plate according to claim 1 or 2, wherein: The hole side wall member has a glass transition temperature of 30° C. or higher as measured by solid viscoelastic temperature dispersion measurement.

12. The orifice plate according to claim 11, wherein The hole side wall member has a glass transition temperature of 120° C. or higher as measured by solid viscoelastic temperature dispersion measurement.

13. The orifice plate according to claim 1 or 2, wherein: The surface free energy of the hole side wall component is 18.0 to 24.3 mN / m. 14 . A method for culturing cells, tissues or organs, comprising the step of culturing cells, tissues or organs using the well plate according to claim 1 or 2.

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