Cell container sheet, cell container, and method for culturing cells

Abstract

To provide a cell container sheet that has high adhesion to other members of a cell container and a high release property.SOLUTION: Provided is a single-layer or multi-layer cell container sheet having an adhesive layer containing a 4-methyl-1-pentene polymer, the peel strength measured by a 90° peel test under the following measurement conditions being 0.010 N / cm or more and 0.100 N / cm or less. (Measurement conditions) A 10 mm×100 mm cell container sheet is pressed against a flat surface of a 1.0 mm thick substrate formed from a 4-methyl-1-pentene homopolymer with a glass transition temperature of 30°C at a temperature of 23°C and a pressure of 1,500 kgw / m2 so that the adhesive layer adheres to the substrate, and held for 5 seconds, after which a 90° peel test is performed using a tensile tester.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a cell container sheet, a cell container, and a method for culturing cells. [Background technology]

[0002] Known cell containers for propagating, growing, storing, and transporting cells include those constructed in such a way that a resin sheet or film is attached to the bottom surface of a cylindrical substrate to form a container shape, or that a resin sheet or film is attached to the top surface of a well to form an airtight container.

[0003] The material of the resin sheet or film to be attached is selected in consideration of oxygen permeability, etc. For example, Patent Document 1 describes that a resin film such as polyethylene terephthalate is preferable. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2016-111975 A Summary of the Invention [Problem to be solved by the invention]

[0005] According to the findings of the present inventors, conventional resin sheets such as polyethylene terephthalate do not have sufficient adhesion to other components of the cell container such as the substrate, and liquid components inside the container may volatilize through the gaps in the resin sheet. As a result, for example, in the case of a cell culture well, liquid components are more likely to volatilize in the peripheral wells than in the central wells, which causes the culture environment to change between the peripheral wells and the central wells, making it difficult to maintain a constant culture environment for each well. In addition, if the resin sheet has low adhesion, liquid components inside the container may leak out of the container through the gaps in the resin sheet when the cell container is vibrated.

[0006] Increasing the adhesiveness of the resin sheet can suppress the volatilization and leakage of the liquid components. However, increasing the adhesiveness makes it difficult to peel the resin sheet from the cell container. As a result, it may take time to peel the resin sheet from the cell container, reducing the efficiency of cell culture, or the resin sheet may not be completely peeled off and may stick to other components of the cell container, reducing the efficiency of reuse of the cell container.

[0007] The present invention has been developed in consideration of the problems with the conventional technology described above, and aims to provide a cell container sheet that has high adhesion to other components of the cell container and also has high peelability, a cell container having such a cell container sheet, and a method for culturing cells using such a cell container. [Means for solving the problem]

[0008] One aspect of the present invention for solving the above problems relates to a cell container sheet according to the following items [1] to

[17] . [1] A single-layer or multi-layer cell container sheet having an adhesive layer containing a 4-methyl-1-pentene polymer, The peel strength measured by a 90° peel test under the following measurement conditions is 0.01 N / cm or more and 0.10 N / cm or less. Sheet for cell containers. (Measurement conditions) A 10mm x 100mm cell container sheet was attached to the flat surface of a 1.0mm thick substrate made of 4-methyl-1-pentene homopolymer with a glass transition temperature of 30°C at a temperature of 23°C and a pressure of 1500kgw / m. 2 The adhesive layer is pressed against the substrate and held for 5 seconds, after which a 90° peel test is performed using a tensile tester. [2] The cell container sheet has a maximum value of the loss tangent (tan δ) of dynamic viscoelasticity in the range of 10°C to 100°C, as determined by dynamic viscoelasticity measurement under conditions of a heating rate of 4°C / min, a frequency of 1.59Hz, and a strain of 0.1%; and The maximum value of the loss tangent is 0.5 or more and 3.5 or less. The cell container sheet according to [1]. [3] The 4-methyl-1-pentene polymer is a copolymer having a structural unit derived from 4-methyl-1-pentene and a structural unit derived from ethylene or propylene. The cell container sheet according to [1] or [2]. [4] The cell container sheet has a thickness of 15 μm or more and 3 mm or less. The cell container sheet according to any one of [1] to [3]. [5] The cell container sheet has an oxygen permeability of 7500 cm at 23°C. 3 / (m 2 ·24h・atm) or more than 150000cm 3 / (m 2 ·24h·atm) or less, The cell container sheet according to any one of [1] to [4]. [6] The cell container sheet has a water vapor permeability of 10 g / (m 2 ·24h) or more 100g / (m 2 -24h or less, The cell container sheet according to any one of [1] to [5]. [7] A medical device with a Sterility Assurance Level (SAL) of 10 as measured in accordance with BS EN556-1:2001. -6 Below is the The cell container sheet according to any one of [1] to [6]. [8] When the cell container sheet is attached to a well plate, the thickness of the periphery of the well is thinner than the thickness of the center of the well. The cell container sheet according to any one of [1] to [7]. [9] A multi-layer sheet having an elastomer layer containing one or more thermoplastic elastomers selected from the group consisting of polyolefin-based elastomers, styrene-based elastomers, polyurethane-based elastomers, polyester-based elastomers, and polyamide-based elastomers. The cell container sheet according to any one of [1] to [8].

[10] The cell container sheet has a maximum value of the loss tangent (tan δ) of dynamic viscoelasticity in the range of 10°C to 100°C and in the range of -70°C to -20°C, as determined by dynamic viscoelasticity measurement under conditions of a heating rate of 4°C / min, a frequency of 1.59Hz, and a strain of 0.1%, and the maximum value of the loss tangent in the range of 10°C or more and 100°C or less is 0.5 or more and 3.5 or less; The maximum value of the loss tangent in the range of -70 ° C or more and -20 ° C or less is 0.05 or more and 1.0 or less. The cell container sheet according to [9].

[11] The ratio of the thickness of the adhesive layer to the thickness of the elastomer layer (adhesive layer / elastomer layer) is 0.9 or more and 5.0 or less; The cell container sheet according to [9] or

[10] .

[12] The elastomer layer is laminated at least partially in contact with the adhesive layer. The cell container sheet according to any one of [9] to

[11] .

[13] The cell container sheet has a tab on the periphery for peeling from the cell container. The cell container sheet according to any one of [1] to

[12] .

[14] The tab is disposed at a position that protrudes from the peripheral portion of another member when the cell container sheet is attached to the other member of the cell container. The cell container sheet according to

[13] .

[15] Having an indication portion indicating the surface to be brought into contact with another member when the cell container is brought into close contact with the other member. The cell container sheet according to any one of [1] to

[14] .

[16] A lid material for a cell container. The cell container sheet according to any one of [1] to

[15] .

[17] The base material of the cell container. The cell container sheet according to any one of [1] to

[16] .

[0009] One aspect of the present invention for solving the above-mentioned problems relates to a cell container as set forth in the following items

[18] to

[19] .

[18] A cell container sheet according to any one of [1] to

[17] , A substrate having a through hole or a recess, The cell container sheet is in close contact with the upper or bottom surface of the through-hole, or the upper surface of the recess. cell container.

[19] The substrate has a plurality of wells; The cell container sheet has a thickness smaller at the periphery of the well than at the center of the well.

[18] A cell container according to the invention.

[0010] One aspect of the present invention for solving the above-mentioned problems relates to a method for culturing cells as described in

[20] below.

[20] A step of forming a cell container by adhering the cell container sheet according to any one of [1] to

[17] to a substrate having a through-hole or a recess; Cultivating cells in the cell container; removing the cell container sheet from the cell container; A method for culturing cells comprising the steps of: Effect of the Invention

[0011] According to the present invention, there are provided a cell container sheet which has high adhesion to other components of the cell container and also has high releasability, a cell container having the cell container sheet, and a method for culturing cells using the cell container. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1A is a schematic perspective view showing the appearance of a single-layer cell container sheet, and FIG. 1B is a schematic perspective view showing the appearance of another single-layer cell container sheet. [Diagram 2] FIG. 2A is a schematic perspective view showing the appearance of a cell container sheet which is a two-layer laminate, and FIG. 2B is a schematic perspective view showing the appearance of another cell container sheet which is a two-layer laminate. [Diagram 3]FIG. 3 is a perspective view of a culture vessel, which is a cell vessel having a cell vessel sheet. [Figure 4] FIG. 4A is a cross-sectional view of a portion of a culture vessel, and FIG. 4B is a cross-sectional view of a portion of a culture vessel showing another embodiment of the culture vessel. [Diagram 5] FIG. 5A is a bottom perspective view showing another culture vessel having a cell vessel sheet, and FIG. 5B is a top perspective view of the culture vessel. [Figure 6] FIG. 6 is a cross-sectional view of a portion of the culture vessel. [Figure 7] FIG. 7 is a schematic diagram showing how the cell container sheet is peeled off to collect nerve tissue. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] 1. Cell container sheet The first embodiment of the present invention relates to a cell container sheet. The cell container sheet may be any member that is in close contact with other members such as the main body (substrate) of the cell container to form a part of the cell container, and may be used as a lid material, a bottom material, a side material, or any of a variety of other members of the cell container.

[0014] 1-1. Shape and composition of cell container sheet The cell container sheet may be a single layer or a multi-layer laminate.

[0015] Fig. 1A is a schematic perspective view showing the appearance of a single-layer cell container sheet 100a. Fig. 1B is a schematic perspective view showing the appearance of another single-layer cell container sheet 100b. The cell container sheet 100a shown in Fig. 1A and the cell container sheet 100b shown in Fig. 1B each consist of a single-layer adhesion layer 110 formed by molding a resin composition containing a 4-methyl-1-pentene polymer into a film or sheet shape.

[0016] Fig. 2A is a schematic perspective view showing the appearance of a cell container sheet 200a which is a two-layer laminate. Fig. 2B is a schematic perspective view showing the appearance of another cell container sheet 200b which is a two-layer laminate. The cell container sheet 200a shown in Fig. 2A and the cell container sheet 200b shown in Fig. 2B have an adhesion layer 210 formed by molding a resin composition containing a 4-methyl-1-pentene polymer into a film or sheet, and an elastomer layer 220 containing a thermoplastic elastomer.

[0017] The cell container sheet may be a laminate of three or more layers, in which case the adhesion layer containing the 4-methyl-1-pentene polymer is disposed at a position that will become the uppermost layer (or the lowermost layer) of the laminate when the cell container sheet is adhered to another member.

[0018] As described below, the cell container sheet according to this embodiment has high adhesion to other components of the cell container and is also easy to peel off. The cell container sheet 100b and the cell container sheet 200b shown in Fig. 1B and Fig. 2B have peeling tabs 112 and 212 on the edge of the sheet to facilitate peeling from the cell container. The tabs 112 and 212 protrude outward from the edge of the cell container sheet. The tabs 112 and 212 can be pinched and pulled to more easily peel the cell container sheet 100b and the cell container sheet 200b from the cell container.

[0019] Tabs 112 and 212 may be formed by molding one or more layers from among the adhesive layer, elastomer layer, and other layers so that the end edges have protrusions in the shape of tabs 112 and 212, or separately manufactured tabs 112 may be attached to the end edges by a method such as heat fusion or adhesion. The number of tabs 112 and 212 is not limited and may be one or more, but one or two is preferable.

[0020] As described later, the cell container sheet according to this embodiment is attached to another member of the cell container by contacting the adhesion layer containing the 4-methyl-1-pentene polymer. The cell container sheet 200b shown in FIG. 2B has an indicator 222 for indicating which side (adhesion layer) is to be in contact with the other member. The indicator 222 may be formed by attaching a sticker indicating which side is the corresponding side to one or both of the outer side (opposite side to the other member) and the inner side (side in contact with the other member). Alternatively, the indicator 222 may be formed by forming a colored area in a portion protruding from the other member to the outside so that letters cannot be seen from the back side, and writing which side is the corresponding side with ink or the like on one or both sides of the area.

[0021] The thickness of the cell container sheet is preferably 15 μm or more and 3 mm or less, more preferably 18 μm or more and 1 mm or less, and even more preferably 20 μm or more and 500 μm or less. By increasing the thickness, the mechanical strength, shape retention, and moisture resistance of the cell container sheet can be increased. By decreasing the thickness, the oxygen permeability, carbon dioxide permeability, water vapor permeability, shape conformability, lightness, handling, transparency, and moldability of the cell container sheet can be increased. The thickness of the cell container sheet can also be determined according to the required characteristics. For example, from the viewpoint of increasing oxygen permeability, it is preferably 15 μm or more and 150 μm or less, and more preferably 20 μm or more and 100 μm or less.

[0022] The shape of the cell container sheet in plan view can be set according to the shape of the other member to which it is adhered. The cell container sheet preferably has a larger area than the surface of the other member to which it is adhered. When such a cell container sheet is adhered to the other member, a part of the cell container sheet protrudes from the peripheral portion of the other member. The protruding portion makes it easier to peel the cell container sheet from the other member, and can improve the peelability of the cell container sheet from the cell container. In addition, tabs 112 and 212, display unit 222, etc. can be formed in the protruding portion that does not affect cell culture, etc. The protruding portion is preferably formed around the entire circumference of the other member to which the cell container sheet is adhered.

[0023] 1-2.Characteristics of cell container sheets The cell container sheet has a peel strength of 0.01 N / cm or more and 0.10 N / cm or less, preferably 0.02 N / cm or more and 0.09 N / cm or less, and more preferably 0.03 N / cm or more and 0.08 N / cm or less, as measured by a 90° peel test performed under the following measurement conditions. By setting the peel strength to 0.010 N / cm or more, the adhesion of the cell container sheet to other components of the cell container can be increased, and the volatilization or leakage of liquid components from the gap between the cell container sheet and other components can be sufficiently suppressed. By setting the peel strength to 0.10 N / cm or less, the cell container sheet can be easily peeled from other components after use of the cell container.

[0024] (Measurement conditions) A 10mm x 100mm cell container sheet was attached to the flat surface of a 1.0mm thick substrate made of 4-methyl-1-pentene homopolymer with a glass transition temperature of 30°C at a temperature of 23°C and a pressure of 1500kgw / m. 2 The adhesive layer is pressed against the substrate and held for 5 seconds, after which a 90° peel test is performed using a tensile tester.

[0025] The peel strength can be adjusted to within the above range by the composition of the 4-methyl-1-pentene polymer and the amount of the 4-methyl-1-pentene polymer in the resin composition that is the adhesive layer material.

[0026] The cell container sheet preferably has at least one maximum value of the loss tangent (tan δ) of dynamic viscoelasticity in the range of 10°C to 100°C, as determined by dynamic viscoelasticity measurement under conditions of a heating rate of 4°C / min, a frequency of 1.59 Hz, and a distortion of 0.1%. If the maximum value of tan δ is in this range, the cell container sheet is likely to become flexible at room temperature or a temperature slightly warmer than this. Therefore, when the cell container sheet is attached to another member at room temperature, the cell container sheet is likely to adhere to the other member. Furthermore, when the cell container sheet is attached to another member while pressing it with the fingers of a hand, the cell container sheet is likely to become moderately flexible and is likely to adhere more closely to the other member. From the above viewpoint, the cell container sheet preferably has a maximum value of tan δ in the range of 20°C to 60°C, and more preferably in the range of 25°C to 40°C.

[0027] In addition, the cell container sheet preferably has a maximum value of tan δ (at least one of the maximum values ​​when there are multiple maximum values) in the above-mentioned range of 10°C to 100°C of 0.5 to 3.5, more preferably 0.6 to 3.0, and even more preferably 0.7 to 2.8. If the maximum value of tan δ in the above-mentioned range of 10°C to 100°C is 0.5 or more, the shape retention of the cell container sheet is likely to be improved, and the sheet is unlikely to be damaged when attached or peeled off. If the maximum value of tan δ in the above-mentioned range of 10°C to 100°C is 3.5 or less, the shape conformability of the cell container sheet is likely to be improved, and the adhesion to other members of the cell container is likely to be improved. In addition, when the cell container sheet is attached to a member having a well, the cell container sheet is likely to deform into a shape that fits inside the well (described later).

[0028] Furthermore, the cell container sheet preferably has at least one maximum value of the loss tangent (tan δ) of dynamic viscoelasticity in the range of -70°C or more and -20°C or less, as determined by dynamic viscoelasticity measurement under conditions of a heating rate of 4°C / min, a frequency of 1.59 Hz, and a distortion of 0.1%. If the cell container sheet has a maximum value of tan δ in this range, the cell container sheet becomes more flexible and easily adheres to other members. From the above viewpoint, the cell container sheet preferably has a maximum value of tan δ in the range of -65°C or more and -20°C or less, and more preferably in the range of -60°C or more and -20°C or less.

[0029] In addition, the cell container sheet preferably has a maximum value of tan δ (at least one of the maximum values ​​when there are multiple maximum values) in the range of -70°C to -20°C of 0.01 to 1.0, more preferably 0.02 to 0.7, and even more preferably 0.03 to 0.5. If the maximum value of tan δ in the range of -70°C to -20°C is 0.05 or more, the shape retention of the cell container sheet is likely to be improved and the sheet is unlikely to be damaged when attached or peeled off. If the maximum value of tan δ in the range of -70°C to -20°C is 1.0 or less, the shape conformability of the cell container sheet is likely to be improved and the adhesion to other members of the cell container is likely to be improved. In addition, when the cell container sheet is attached to a member having a well, the cell container sheet is likely to deform into a shape that fits inside the well (described later).

[0030] In addition, the cell container sheet has an oxygen permeability of 7500 cm at 23°C. 3 / (m 2 ·24h・atm) or more than 150000cm 3 / (m 2 24h atm) or less, and 3 / (m 2 ·24h・atm) or more than 60000cm 3 / (m 2 24h atm) or less, and 8500cm 3 / (m 2 ·24h・atm) or more than 40000cm 3 / (m2 It is more preferable that the oxygen permeability is less than 100% (24h·atm). The higher the oxygen permeability, the easier it is to promote cell proliferation and growth. If the oxygen permeability is appropriately low, cell adhesion will be good, making it easier to apply the cell container sheet when culturing a wider variety of cells.

[0031] The oxygen permeability was measured using a differential pressure gas permeability measuring device (MT-C3, manufactured by Toyo Seiki Seisakusho) at a temperature of 23°C and humidity of 0%. 3 / (m 2 The measured value was 90 × 90 mm test pieces cut out from the cell container sheet, and the measurement area was 70 mm (the transmission area was 38.46 cm). 2 When the oxygen permeability is expected to be large, an aluminum mask is placed on the measurement sample in advance to reduce the actual permeation area to 5.0 cm. 2 It is more preferable to set the above.

[0032] The cell container sheet also has a water vapor permeability of 10 g / (m2) at 40°C and a relative humidity of 90%. 2 ·24h) or more 100g / (m 2 24h) or less, and 15g / (m 2 ·24h) or more 80g / (m 2 24h) or less is more preferable, and 20g / (m 2 ·24h) or more 70g / (m 2 When the water vapor transmission rate is within the above range, the medium is less likely to dry out during long-term culture.

[0033] The water vapor permeability above is for a surface area of ​​50 cm due to heat fusion of the ends. 2 The cell container sheet is processed into a bag shape and calcium chloride is enclosed inside as a moisture absorbent. The cell container sheet is then placed in a thermohygrostat at 40°C and 90% RH, and the mass is repeatedly measured every 24 hours. After confirming that the amount of mass change has become linear on the graph, the amount of mass change per 24 hours (1 day) (g / 24h) is calculated from the linear portion, and the amount of mass change is multiplied by the surface area (m2 The value obtained by dividing the calculated value by the water vapor permeability of the cell container sheet is defined as the water vapor permeability of the cell container sheet.

[0034] In addition, the cell container sheet has a sterility assurance level (SAL) of 10 for medical devices measured in accordance with BS EN556-1:2001. -6 It is preferable that:

[0035] The cell container sheet is preferably transparent from the viewpoint of improving the efficiency of cell observation. Specifically, the total light transmittance measured in accordance with JIS K 7361-1 is preferably 50% or more and 100% or less, and more preferably 70% or more and 100% or less.

[0036] 1-3. Adhesion layer The adhesion layer is a layer containing a 4-methyl-1-pentene polymer. The adhesion layer containing a 4-methyl-1-pentene polymer can improve the adhesion of the cell container sheet to other members and oxygen permeability.

[0037] The thickness of the adhesion layer is preferably 10 μm or more and 0.9 mm or less, more preferably 15 μm or more and 0.6 mm or less, and even more preferably 18 μm or more and 100 μm or less. The thicker the adhesion layer, the higher the adhesion of the cell container sheet to other members. If the adhesion layer is appropriately thin, the oxygen permeability of the cell container sheet becomes sufficiently high.

[0038] The 4-methyl-1-pentene polymer may be a homopolymer of 4-methyl-1-pentene or a copolymer of 4-methyl-1-pentene and another α-olefin (excluding 4-methyl-1-pentene). The other α-olefin is preferably an α-olefin having 2 to 20 carbon atoms.

[0039] The other α-olefin may be a linear olefin, a branched olefin, or a cyclic olefin.

[0040] The other linear α-olefin is more preferably an α-olefin having 2 to 10 carbon atoms, and even more preferably an α-olefin having 2 to 3 carbon atoms. Examples of the other linear α-olefin include ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, 1-decene, 1-undecene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, and 1-eicosene. Of these, ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, and 1-decene are preferred, and ethylene and propylene are more preferred.

[0041] From the viewpoint of enhancing the flexibility and mechanical strength of the adhesive layer, the 4-methyl-1-pentene polymer preferably has an intrinsic viscosity [η] in decalin at 135°C of 0.01 dL / g or more and 5.0 dL / g or less, more preferably 0.1 dL / g or more and 4.0 dL / g or less, even more preferably 0.5 dL / g or more and 3.0 dL / g or less, and particularly preferably 1.0 dL / g or more and 2.8 dL / g or less.

[0042] The density of the 4-methyl-1-pentene polymer measured in accordance with ASTM D1505 (underwater displacement method) is 0.810 g / cm 3 More than 0.850g / cm 3 It is preferable that the density is 0.820 g / cm or less. 3 More than 0.850g / cm 3 More preferably, it is 0.830 g / cm or less. 3 More than 0.850g / cm 3 It is even more preferable that:

[0043] The 4-methyl-1-pentene polymer can be produced by a known method using a known catalyst such as a magnesium-supported titanium catalyst, a metallocene catalyst (WO 01 / 53369, WO 01 / 027124, JP 3-193796 A, and JP 02-41303 A, etc.), or an olefin polymerization catalyst containing a metallocene compound (WO 2011 / 055803 A).

[0044] The adhesion layer may contain additives such as a crosslinking agent, a crosslinking auxiliary, a foaming agent, a decomposition temperature regulator, a weather resistance stabilizer, a heat resistance stabilizer, an antistatic agent, an antislip agent, an antiblocking agent, an antifogging agent, a lubricant, a pigment, a dye, a plasticizer, an antioxidant, a hydrochloric acid absorber, an antioxidant, a metal damage inhibitor, a crystal nucleating agent, an antifungal agent, an antibacterial agent, a flame retardant, a filler (inorganic filler, organic filler), and a softener.

[0045] The content of the 4-methyl-1-pentene polymer in the adhesive layer is preferably 50% by mass or more and 100% by mass or less, more preferably 50% by mass or more and 99.5% by mass or less, even more preferably 60% by mass or more and 99% by mass or less, even more preferably 65% ​​by mass or more and 98% by mass or less, and particularly preferably 70% by mass or more and 97% by mass or less. The content of the 4-methyl-1-pentene polymer can be appropriately determined depending on the flexibility, shape retention, shape conformability, light weight, mechanical properties, handleability, appearance, moldability, moisture resistance, etc. of the adhesive layer.

[0046] 1-4.Elastomer layer The elastomer layer is a layer containing one or more thermoplastic elastomers selected from polyolefin-based elastomers, styrene-based elastomers, polyurethane-based elastomers, polyester-based elastomers, and polyamide-based elastomers. The elastomer layer can improve the adhesion and shape retention of the cell container sheet to other members. In this specification, the thermoplastic elastomer refers to a polymer that exhibits thermoplastic properties when heated to a melting point or higher, but exhibits rubber elasticity at room temperature.

[0047] The thickness of the elastomer layer is preferably 10 μm or more and 0.5 mm or less, more preferably 15 μm or more and 0.3 mm or less, and even more preferably 20 μm or more and 100 μm or less. The thicker the elastomer layer, the higher the adhesion of the cell container sheet to other members. If the thickness of the elastomer layer is appropriately thin, the oxygen permeability of the cell container sheet becomes sufficiently high.

[0048] The ratio of the thickness of the adhesion layer to the thickness of the elastomer layer (adhesion layer / elastomer layer) is preferably 0.9 to 5.0, more preferably 0.95 to 4.0, and even more preferably 0.98 to 3.0. The higher the ratio of the adhesion layer, the higher the oxygen permeability of the cell container layer sheet can be.

[0049] Examples of polyolefin elastomers include copolymers of ethylene or propylene with butadiene, isoprene, isobutylene or α-olefins, or hydrogenated copolymers thereof; and blends of polyethylene or polypropylene with ethylene-propylene copolymers, ethylene-propylene-diene copolymers, ethylene-butene copolymers, or hydrogenated styrene-butadiene copolymers (which may be crosslinked).

[0050] Examples of the polyolefin elastomer copolymer include block copolymers of a polyolefin block that is a hard part formed from a highly crystalline polymer such as polypropylene, and a block that is a soft part formed from a monomer copolymer exhibiting amorphous properties. Examples of the block copolymer include olefin (crystalline)-ethylene-butylene-olefin block copolymer, and polypropylene-polyolefin (amorphous)-polypropylene block copolymer.

[0051] Examples of the above styrene-based elastomers include block copolymers (SBS) of a polystyrene block that forms the hard portion (crystalline portion) and a diene monomer block that forms the soft portion, hydrogenated styrene-butadiene-styrene block copolymers (HSBR), styrene-ethylene-propylene-styrene block copolymers (SEPS), styrene-ethylene-butene-styrene block copolymers (SEBS), styrene-isoprene-styrene block copolymers (SIS), styrene-isobutylene-styrene copolymers (SIBS), and styrene-isobutylene copolymers (SIB).

[0052] Examples of the polyester elastomer include block copolymers of aromatic polyester and aliphatic polyester, and block copolymers of aromatic polyester and aliphatic polyether. The aromatic polyester is preferably a polybutylene terephthalate resin or a polyethylene terephthalate resin. The aliphatic polyether is preferably a polyalkylene glycol resin, more preferably a polytetramethylene glycol resin or a polyethylene glycol resin.

[0053] Examples of the polyamide-based elastomer include polyamide resins such as aliphatic polyamides (nylon 6, nylon 11, nylon 12, nylon 66, nylon 610, nylon 612), block copolymers of polyamides and aliphatic polyesters, and block copolymers of polyamides and aliphatic polyethers.

[0054] Among the above thermoplastic elastomers, from the viewpoint of increasing the oxygen permeability of the cell container sheet, styrene-based elastomers are preferred, and hydrogenated styrene-based elastomers such as SEPS and SEBS are more preferred. Also, from the viewpoint of increasing the oxygen permeability of the cell container sheet, styrene-based elastomers with a low styrene content are preferred. For example, the styrene content of the styrene-based elastomer can be 5% by mass or more and 60% by mass or less, and is preferably 7% by mass or more and 30% by mass or less.

[0055] The elastomer layer is preferably disposed (laminated) at least partially in contact with the adhesive layer. When the cell container sheet is in close contact with another member of the cell container, it may be a laminate of two layers, an adhesive layer and an elastomer layer.

[0056] 1-5.Other layers The cell container sheet may have layers other than the adhesive layer and the elastomer layer. For example, the cell container sheet may have a peelable protective layer in contact with the adhesive layer or the elastomer layer for protecting the adhesive layer or the elastomer layer before use. These protective layers may have one or more tabs for peeling the protective layer. The above-mentioned display unit may be formed on the protective layer.

[0057] 2. Cell container and method for culturing cells The above-mentioned cell container sheet can be attached closely to another member to form a part of the cell container.

[0058] 3 is a perspective view of a culture vessel 300, which is a cell vessel having a cell vessel sheet. FIG. 4A is a cross-sectional view of a portion of the culture vessel 300.

[0059] In this specification, culturing a cell means allowing the cell to grow, proliferate, or maintain the cell in a living state.

[0060] The culture vessel 300 is used, for example, for culturing cells derived from a human. Such a culture vessel 300 has a plurality of storage sections 310 (see FIG. 4A) for culturing cells. The culture vessel 300 is accommodated in a culture space of a culture device (for example, an incubator) with a drug (culture solution) and cells to be cultured (hereinafter referred to as "target cells") accommodated in the storage sections 310.

[0061] The culture vessel 300 can be suitably used for culturing spheroids (cell aggregates) of the target cells. The culture vessel 300 does not need to be used in a state housed in a culture device. The culture vessel 300 may be used in various situations depending on the target cells.

[0062] The culture container 300 has a base material 320 and a cell container sheet 330 .

[0063] The substrate 320 has a frame 322 and a plurality of wells 324. The substrate 320 is made of, for example, a synthetic resin, and is an integrally molded product produced by injection molding. Examples of synthetic resins constituting the substrate 320 include polystyrene and polyolefin. Examples of the polyolefin include a cyclic olefin (co)polymer, a 4-methyl-1-pentene (co)polymer, polypropylene, and polyethylene. Among these, from the viewpoint of increasing the gas permeability of the substrate 320, a 4-methyl-1-pentene polymer is preferable. Alternatively, from the viewpoint of increasing the heat resistance of the substrate 320, a 4-methyl-1-pentene polymer, polystyrene, polypropylene, and a cyclic olefin polymer are preferable. As the 4-methyl-1-pentene polymer, the 4-methyl-1-pentene polymer described as the material of the adhesion layer of the cell container sheet can be used in the same manner, but a polymer different from the material of the adhesion layer may be used.

[0064] The frame portion 322 is configured of a rectangular frame-shaped member. The well 324 is provided in a space surrounded by the frame portion 322. The well 324 is provided integrally with the frame portion 322.

[0065] The wells 324 are cylindrical and open upward. In this embodiment, the shape of the opening of the wells 324 (in other words, the outer shape in a plan view) is circular. The wells 324 are arranged side by side in the left-right direction and the front-rear direction. The number of wells 324 may be, for example, 6, 24, 96, or 384. Of course, the number of wells 324 may be other than 6, 24, 96, and 384.

[0066] Adjacent wells 324 are connected to each other by connecting parts 326. The left end well 324, the right end well 324, the front end well 324, and the rear end well 324 are connected to the inner circumferential surface of frame part 322 via connecting parts (not shown).

[0067] The base material 320 has a bottom plate portion 328 on its bottom surface. The bottom plate portion 328 is a plate-shaped member that constitutes the bottom surface of the wells 324 and connects the lower ends of the wells 324 together. The outer edge portion of the bottom plate portion 328 is connected to the inner circumferential surface of the frame portion 322 around the entire periphery.

[0068] The cell container sheet 330 is a lid material for the culture container 300. The cell container sheet 330 seals the inside of the well 324, which is a recess, and suppresses the volatilization of liquid components such as the culture solution and the leakage of liquid components from the well 324 due to vibration of the culture container 300, etc.

[0069] The cell container sheet 330 may be a single-layer sheet as shown in Figures 1A and 1B, or a two-layer (multi-layer) sheet as shown in Figures 2A and 2B. In Figure 3, the cell container sheet 200b having the tab 212 and the display unit 222 as shown in Figure 2B is used as the cell container sheet 330.

[0070] The cell container sheet 330 is placed on the upper surface of the substrate 320 (the upper surface of the well 324) and pressed to adhere to the upper surface of the substrate 320 (the upper surface of the well 324) (a process for forming a cell container). The method of pressing is not particularly limited, and the cell container sheet 330 may be pressed against the upper surface of the substrate 320 with a human finger, or the cell container sheet 330 may be pressed against the upper surface of the substrate 320 using a member such as a roll.

[0071] The cell container sheet 330 having the above-mentioned characteristics is slightly deformed by the above-mentioned pressure, and penetrates slightly into the well 324 as shown in Fig. 4A. At this time, the thickness T1 of the cell container sheet 330 at the periphery of the well 324 stretched by the pressure becomes thinner than the thickness T2 at the center of the well 324. By adopting such a configuration that generates negative pressure due to the volumetric change of the well 324 when the cell container sheet 330 is peeled off, the adhesion of the cell container sheet 330 to the well 324 can be more sufficiently improved.

[0072] On the other hand, by pressing the cell container sheet 330 with an appropriate strength, it can be adhered to the base material 320 to such an extent that it can be easily peeled off (the step of removing the cell container sheet) by hand after cell culture, etc. (the step of culturing cells). The strength of the pressure at the time of adhesion is 300 kgw / m 2 More than 8000kgw / m 2 It is preferable that the weight is less than 400 kgw / m 2 More than 7000kgw / m 2 It is more preferable that the pressure applied is equal to or less than the above. The pressure applied is the normal pressure applied by a human finger. In this manner, the cell container sheet 330 can be very easily attached to the base material 320 (or other cell container components) to enhance the sealing performance and to an extent that the sheet can be easily peeled off.

[0073] FIG. 4B is a cross-sectional view of a portion of the culture vessel 300 shown in FIG. 3, showing another embodiment of the culture vessel.

[0074] 4B, the culture vessel 300 has a well 324 in the form of a through hole in the base material 320. A cell container sheet 330a as a cover material is attached to one opening (upper surface) of the well 324, and a cell container sheet 330b as a base material is attached to the other opening (lower surface). Each of the cell container sheets 330a and 330b slightly extends into the well 324, and a thickness T1 at the periphery of the well 324 is thinner than a thickness T2 at the center of the well 324.

[0075] In the configuration shown in FIG. 4B, the above-mentioned cell container sheet may be adhered to only one of the openings of the well 324, and another resin sheet may be adhered or bonded to the other opening.

[0076] Fig. 5A is a bottom perspective view showing another culture vessel 500 having a cell container sheet. Fig. 5B is a top perspective view of the culture vessel 500. Fig. 6 is a cross-sectional view of a portion of the culture vessel 500.

[0077] The culture vessel 500 is used, for example, for culturing nerve cells. Such a culture vessel 500 has, for example, chambers 524a and 524b in which cell bodies 540 of ungrown nerve cells induced and differentiated from stem cells are placed, and a flow path 524c that communicates with the chambers 524a and 524b. The culture vessel 500 is accommodated in a culture space of a culture device (for example, an incubator) in a state in which a drug (culture solution) is accommodated in the chambers 524a, 524b, and the flow path 524c, and a mass of cell bodies 540 of nerve cells, which are target cells to be cultured, is accommodated in one of the chambers 524a and 524b.

[0078] The cell bodies 540 housed in the chamber each grow an axon 542 along the channel 524c. At this time, the axons 542 self-organize and extend into bundles, forming nerve tissue (a process for culturing cells).

[0079] The culture container 500 has a base material 520 and a cell container sheet 530 .

[0080] Base material 520 has frame portion 522 and bottom plate portion 528. At positions of bottom plate portion 528 corresponding to chamber 524a, chamber 524b, and flow path 524c, through holes are formed in the shapes of chamber 524a, chamber 524b, and flow path 524c. Base material 520 is an integrally molded product made by injection molding from the same synthetic resin as base material 320, for example.

[0081] 5A, the cell container sheet 530 is placed on the bottom surface of the substrate 520 (the bottom surfaces of the chambers 524a, 524b, and flow paths 524c) and pressed to adhere to the bottom surface of the substrate 520 (a process for forming a cell container). The method of pressing is not particularly limited, and the cell container sheet 530 may be pressed against the bottom surface of the substrate 520 with a human finger, or the cell container sheet 530 may be pressed against the bottom surface of the substrate 520 using a member such as a roll. The strength of the pressing may be as described above.

[0082] This forms a culture vessel 500 having a cell container sheet 530 as a base material for the chambers 524a, 524b, and flow channel 524c. At this time, the cell container sheet 530 slightly enters the inside of the through-hole, and the thickness T1 at the periphery of the through-hole is thinner than the thickness T2 at the center of the through-hole.

[0083] The cell container sheet 530 may be a single-layer sheet as shown in Figures 1A and 1B, or a two-layer (multi-layer) sheet as shown in Figures 2A and 2B. In Figures 5A, 5B, and 6, the cell container sheet 200b having the tab 212 and the display unit 222 as shown in Figure 2B is used as the cell container sheet 530.

[0084] In a conventional culture vessel for culturing nerve cells, water pressure is applied to chamber 524a and chamber 524b (the chambers that do not contain cell bodies 540) to push grown nerve tissue out of the chamber from the cell bodies 540 side, and the nerve tissue is collected. However, at this time, the nerve tissue is easily damaged by water pressure or contact with the chamber or flow channel.

[0085] In contrast, the culture container 500 of this embodiment can easily peel off the cell container sheet 530, which is the base material (step of removing the cell container sheet). Then, as shown in Fig. 7, by peeling off the cell container sheet 530, the nerve tissue 544 on the cell container sheet 530 can be easily collected without damage.

[0086] As shown in Figures 3, 5A, 5B, 6 and 7, in this embodiment, the area of ​​the cell container sheet is larger than the area of ​​the surface of the substrate to which the cell container sheet is adhered. The cell container sheet extends beyond the peripheral edge of the substrate over the entire circumference of the substrate. This can improve the ease of peeling the cell container sheet from the cell container.

[0087] 3. Other embodiments It should be noted that each of the above-described embodiments shows an example of the present invention, and the present invention is not limited to the above-described embodiments. Needless to say, various other embodiments are possible within the scope of the concept of the present invention.

[0088] For example, the above-mentioned cell container is a culture container for growing or growing cells, but a container for storing or transporting cells or for observing cells may be a cell container having a cell container sheet. Furthermore, the cells targeted by the cell container are not particularly limited, and may be differentiated somatic cells or undifferentiated stem cells. The cells may be living cells or dead cells. EXAMPLES

[0089] The present invention will be described in detail based on examples, but the present invention is not limited to these examples.

[0090] 1. Preparation of materials 1-1.4-Methyl-1-pentene polymer 1-1-1. 4-Methyl-1-pentene polymer-3 (4MP1-3) The proportion of structural units derived from 4-methyl-1-pentene is 84 mol%, the proportion of structural units derived from propylene is 16 mol%, the intrinsic viscosity [η] is 1.4 dL / g, and the density is 0.838 g / cm 3 The 4-methyl-1-pentene-propylene copolymer with a melt flow rate (230°C, 2160 g) of 10 g / 10 min was named 4-methyl-1-pentene polymer-3 (4MP1-3).

[0091] 1-1-2.4-Methyl-1-pentene polymer-4 (4MP1-4) The proportion of structural units derived from 4-methyl-1-pentene is 72 mol %, the proportion of structural units derived from propylene is 28 mol %, the intrinsic viscosity [η] is 1.5 dL / g, and the density is 0.840 g / cm 3The 4-methyl-1-pentene-propylene copolymer having a melt flow rate (230°C, 2160 g) of 10 g / 10 min was named 4-methyl-1-pentene polymer-4 (4MP1-4).

[0092] 1-1-3. 4-Methyl-1-pentene polymer-1 (4MP1-1) The mixture obtained by mixing 60 parts by mass of 4MP1-3 and 40 parts by mass of 4MP1-4 was named 4-methyl-1-pentene polymer-1 (4MP1-1). 4MP1-1 had 79 mol% of structural units derived from 4-methyl-1-pentene and 21 mol% of structural units derived from propylene.

[0093] 1-1-4. 4-Methyl-1-pentene polymer-2 (4MP1-2) The mixture obtained by mixing 80 parts by mass of 4MP1-3 and 20 parts by mass of 4MP1-4 was named 4-methyl-1-pentene polymer-2 (4MP1-2). 4MP1-2 had 82 mol% of structural units derived from 4-methyl-1-pentene and 18 mol% of structural units derived from propylene.

[0094] The intrinsic viscosity [η] of the 4-methyl-1-pentene polymer is a value obtained from the measurement results using an Ubbelohde viscometer. Specifically, about 20 mg of the polymer was dissolved in 25 ml of decalin, and the specific viscosity η sp The decalin solution was diluted with 5 ml of decalin, and the specific viscosity η sp The dilution and specific viscosity η sp The measurement was repeated two more times to obtain the concentration (C) and specific viscosity η sp The relational equation showing the relationship between the concentration (C) and the concentration (C) was obtained. In the above relational equation, η sp The value of / C was taken as the intrinsic viscosity [η] (unit: dl / g).

[0095] The density of the 4-methyl-1-pentene polymer is a value determined in accordance with ASTM D1505 (density gradient method).

[0096] The melt flow rate (MFR) of the 4-methyl-1-pentene polymer is a value measured at 230° C. under a load of 2160 g in accordance with ASTM D1238.

[0097] 1-2. Thermoplastic elastomer SEBS (Kraton G1657M, Kraton Corporation) with a styrene content of 13% by mass was used as the thermoplastic elastomer.

[0098] 2. Preparation of cell container sheets 2-1. Preparation of Sheet 1 Using the materials listed in Table 1, a co-extrusion film molding machine having a screw with a diameter of 50 mm and an effective length L / D=28 was used, with the cylinder temperature set to 100-230°C and the die temperature set to 130-230°C, and the molten mixture of 4MP1-1 (adhesive layer) and the molten mixture of SEBS (elastomer layer) were co-extruded from the T-die to thicknesses of 50 μm (adhesive layer) and 50 μm (elastomer layer), respectively, and cast-molded using a mirror-finished roll set at 20°C to obtain Sheet 1 (thickness: 100 μm). The thickness of Sheet 1 was 50 μm for the adhesive layer and 50 μm for the elastomer layer, totaling 100 μm.

[0099] 2-2. Preparation of Sheets 2 to 5 Sheets 2 to 5 were obtained in the same manner as in the preparation of Sheet 1, except that the type and thickness configuration of the resin material used in preparing the adhesive layer and elastomer layer were changed as shown in Table 1.

[0100] 2-3. Preparation of Sheet 6 Using a molding device having a single-screw extruder with a cylinder inner diameter D of 65 mm, a full-flight screw, and a T-die with a die width of 750 mm, a cooling roll, and a take-off machine, 4MP1-3 was put into the single-screw extruder, melt-kneaded at temperatures of 100°C to 250°C and a screw rotation speed of 23 rpm, and extruded from the T-die so that the extrusion rate was 15 kg / hour. The extruded sheet was cooled by a cooling roll with a water temperature of 20°C in the roll, and taken up by a take-off machine at a take-up speed of 1.3 m / min to obtain a single-layer sheet 6 having an adhesive layer containing 4MP1-3. The thickness of the sheet 6 was 300 μm and the sheet width was 700 mm.

[0101] 2-4. Preparation of Sheet 7 Except for changing the type of resin material to 4MP1-4, the production was performed in the same manner as for the sheet 6, to obtain a single-layer sheet 7 having an adhesive layer containing 4MP1-4. The thickness of the sheet 7 was 300 μm.

[0102] 2-9. Preparation of Sheet 8 A commercially available PET film (thickness 80 μm, manufactured by BM Kiki Co., Ltd., product name: Sureseal ST) having an adhesive layer and a base layer was used as sheet 8.

[0103] 3. Measurement For each sheet, the maximum value of loss tangent (tan δ), oxygen permeability, water vapor permeability and peel strength were measured by the following methods.

[0104] 3-1. Maximum value of loss tangent (tan δ) (dynamic viscoelasticity measurement) Each sheet was cut into a size of 5 mm wide and 30 mm long to prepare a test piece. From the results of dynamic viscoelasticity measurements performed under the following conditions using a solid dynamic viscoelasticity measuring device, the temperature at which the maximum value of the loss tangent (tan δ), which is the ratio (E'' / E') of the storage modulus (E') to the loss modulus (E''), appears and the value of this maximum value were determined. (Measurement conditions) Measurement equipment: RSA-III, manufactured by TA Instruments Co., Ltd. Deformation mode: tension Temperature range: -150℃~150℃ Heating rate: 4℃ / min Frequency: 1.59Hz Distortion: 0.1% Atmosphere: Nitrogen atmosphere

[0105] 3-2.Oxygen permeability Each sheet was cut into a size of 90 mm wide x 90 mm long to prepare a test piece. The circular measurement area of ​​the test piece was 70 mm in diameter (transmission area was 38.46 cm 2 ) was measured at a temperature of 23°C and humidity of 0% using a differential pressure gas permeability measuring device (Toyo Seiki Co., Ltd., MT-C3). 3 / (m 2 ×24h×atm)] was measured.

[0106] 3-3.Water vapor permeability Each sheet is heat-sealed at the edges to a surface area of ​​50 cm 2 The sheet was processed into a bag shape, and calcium chloride was enclosed as a moisture absorbent inside the bag-shaped sheet. The sheet processed into a bag shape and containing calcium chloride was placed in a thermohygrostat at 40°C and a relative humidity of 90%, and the mass was repeatedly measured every 24 hours. After confirming that the amount of mass change formed a straight line on the graph, the amount of mass change per 24 hours (1 day) (g / 24h) was calculated from the linear portion, and the amount of mass change was calculated as a function of the surface area (m2) of the sheet. 2 ) to obtain the water vapor permeability of each sheet.

[0107] 3-4. Peel strength Each sheet was cut into a size of 10 mm wide x 100 mm long to prepare a test piece. Each test piece was attached to the flat surface of a 1.0 mm thick substrate made of 4-methyl-1-pentene homopolymer with a glass transition temperature of 30°C at a temperature of 23°C and a pressure of 1500 kgw / m 2 The adhesive layer was pressed against the substrate and held for 5 seconds, after which a 90° peel test was performed using a tensile tester. The gripping speed was 50 mm / min, and the average peel force over a moving distance of 30 mm to 80 mm was taken as the peel strength (N / 10 mm) of the sheet.

[0108] The configurations and measurement results of Sheets 1 to 8 are shown in Tables 1 and 2.

[0109] [Table 1]

[0110] [Table 2]

[0111] 4. Cell Container Fabrication and Evaluation 4-1. Preparation of cell container 4-1-1. Preparation of cell container 1 A transparent 384-well plate (wells have a square shape when viewed from above) made from 4-methyl-1-pentene polymer was prepared, and sheet 1 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, a finger was pressed against the sheet 1 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 1 to the top surface of the wells, thereby forming cell container 1.

[0112] 4-1-2. Preparation of cell container 2 A transparent 384-well plate made of a styrene polymer and rectangular in plan view (wells have a square shape in plan view) was prepared, and sheet 1 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, a finger was pressed against the sheet and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering sheet 1 to the top surface of the wells, to form cell container 2.

[0113] 4-1-3. Preparation of cell container 3 A transparent 384-well plate made of a styrene polymer and rectangular in plan view (wells have a square shape in plan view) was prepared, and sheet 2 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, a finger was pressed against the sheet 2 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 2 to the top surface of the wells, to form a cell container 3.

[0114] 4-1-4. Preparation of cell container 4 A transparent 384-well plate made of a styrene polymer and rectangular in plan view (wells have a square shape in plan view) was prepared, and sheet 3 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, a finger was pressed against the sheet 3 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 3 to the top surface of the wells, to form a cell container 4.

[0115] 4-1-5. Preparation of cell container 5 A transparent 384-well plate made of a styrene polymer and rectangular in plan view (wells have a square shape in plan view) was prepared, and sheet 4 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, a finger was pressed against the sheet 4 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 4 to the top surface of the wells, to form a cell container 5.

[0116] 4-1-6. Preparation of cell container 6 A transparent 384-well plate made of a styrene polymer and rectangular in plan view (wells have a square shape in plan view) was prepared, and sheet 5 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, a finger was pressed against the sheet 5 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 5 to the top surface of the wells, to form a cell container 6.

[0117] 4-1-7. Preparation of cell container 7 A transparent 384-well plate (wells have a square shape when viewed from above) made from 4-methyl-1-pentene polymer was prepared, and sheet 1 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, a roll weighing 0.32 kg and having a width of 80 mm was moved from one short side of the well plate to the other short side while pressing against the sheet, thereby adhering sheet 1 to the top surface of the wells, thereby forming cell container 7.

[0118] 4-1-8. Preparation of cell container 8 A transparent 384-well plate (wells have a square shape when viewed from above) made from 4-methyl-1-pentene polymer was prepared, and sheet 1 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, the sheet 1 was moved diagonally from one vertex of the well plate to the other vertex while lightly pressing the sheet with fingers, thereby adhering the sheet 1 to the top surface of the wells, to form a cell container 8.

[0119] 4-1-9. Preparation of cell container 9 A transparent 384-well plate (wells have a square shape when viewed from above) made from 4-methyl-1-pentene polymer was prepared, and sheet 1 was placed on the top surface of the wells so that the elastomer layer was on the well side. Then, at 23°C, the sheet 1 was moved diagonally from one vertex of the well plate to the other vertex while pressing firmly with a finger, thereby adhering the sheet 1 to the top surface of the wells, to form a cell container 9.

[0120] 4-1-10. Preparation of cell container 10 A transparent 384-well plate (wells have a square shape when viewed from above) made of a 4-methyl-1-pentene polymer and rectangular in plan view was prepared, and sheet 6 was placed on the top surface of the wells. Then, at 23°C, a finger was pressed against the sheet 6 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 6 to the top surface of the wells, thereby forming a cell container 10.

[0121] 4-1-11. Preparation of cell container 11 A transparent 384-well plate (wells have a square shape when viewed from above) made of a 4-methyl-1-pentene polymer was prepared, and sheet 7 was placed on the top surface of the wells. Then, at 23°C, a finger was pressed against the sheet 7 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 7 to the top surface of the wells, thereby forming a cell container 11.

[0122] 4-1-12. Preparation of cell container 12 A transparent 384-well plate (wells have a square shape when viewed from above) made of a 4-methyl-1-pentene polymer and rectangular in plan view was prepared, and sheet 9 was placed on the top surface of the wells. Then, at 23°C, a finger was pressed against the sheet 9 and moved diagonally from one vertex of the well plate to the other vertex, thereby adhering the sheet 9 to the top surface of the wells, thereby forming a cell container 12.

[0123] The strength of the fingers when pressed tightly against the skin (cell container 7), when pressed lightly (cell container 8), and when pressed firmly (cell container 9) was determined by measuring the area of ​​the test subject's hand in advance and then dividing the strength obtained by placing the hand on a weighing scale by the area of ​​the hand.

[0124] 4-2.Evaluation 4-2-1. Seat insertion After being attached to the well plate, the sheet was visually observed from the side of the well plate to check whether the sheet had penetrated into the wells.

[0125] 4-2-3. Airtightness After pouring 300 μl of pure water into each well, the sheet was attached to the top surface of the well by the above method. After attachment, the cell container was left to stand at 23°C for 1 hour, and then the cell container was turned over at 180°C several times, and visually checked for water leakage, and the peelability was evaluated according to the following criteria. ◎ Repeated 180 degree inversion 5 times, but no leakage occurred ○ No leakage occurred after 180 degree inversion three times, but leakage occurred from the fourth inversion onwards × It leaked after just one 180 degree inversion

[0126] 4-2-4. Peelability The edge of the sheet that was attached to the top surface of the well was grasped by hand, and the sheet was peeled off by hand. Based on the ease of peeling, the peelability was evaluated according to the following criteria. ◎ It was easy to peel off ○ I was able to peel it off, but it felt like it was stuck. × Unable to peel off

[0127] Tables 3 and 4 show the methods for producing cell containers 1 to 12 and the evaluation results.

[0128] [Table 3]

[0129] [Table 4]

[0130] The results shown in Tables 3 and 4 indicate that cell container sheets having a peel strength of 0.010 N / cm or more and 0.100 N / cm or less as measured by the 90° peel test can achieve both adhesion and releasability. [Industrial Applicability]

[0131] The cell container sheet and the cell container having the same according to the present invention can be applied to the culture of various cells. [Explanation of symbols]

[0132] 100a, 100b, 200a, 200b Cell container sheet 110, 210 Adhesion layer 112, 212 tabs 220 Elastomer layer 222 Display section 300 Culture vessels 310 Storage unit 320 Base material 322 Frame 324 well 326 Connection 328 Bottom plate part 330, 330a, 330b Cell container sheet 500 culture vessels 520 Base material 522 Frame 528 Bottom plate part 524a, 524b chambers 524c Flow path 530 Cell Container Sheet 540 cell body 542 Axon 544 Nervous Tissue

Claims

1. A single-layer or multi-layer cell container sheet having an adhesive layer containing a 4-methyl-1-pentene polymer, The peel strength measured by a 90° peel test under the following measurement conditions is 0.01 N / cm or more and 0.10 N / cm or less. Sheet for cell containers. (Measurement conditions) A cell container sheet of 10 mm×100 mm was attached to the flat surface of a 1.0 mm thick substrate made of 4-methyl-1-pentene homopolymer with a glass transition temperature of 30° C. at a temperature of 23° C. and a pressure of 1500 kgw / m ２ The adhesive layer is pressed against the substrate with a pressure of 0.15 bar so that the adhesive layer adheres to the substrate, and the pressure is maintained for 5 seconds. Thereafter, a 90° peel test is performed using a tensile tester.

2. The cell container sheet has a maximum value of loss tangent (tan δ) of dynamic viscoelasticity in the range of 10° C. to 100° C., the maximum value being determined by dynamic viscoelasticity measurement under conditions of a temperature rise rate of 4° C. / min, a frequency of 1.59 Hz, and a distortion amount of 0.1%, and The maximum value of the loss tangent is 0.5 or more and 3.5 or less. The cell container sheet according to claim 1 .

3. The 4-methyl-1-pentene polymer is a copolymer having a structural unit derived from 4-methyl-1-pentene and a structural unit derived from ethylene or propylene. The cell container sheet according to claim 1 .

4. The cell container sheet has a thickness of 15 μm or more and 3 mm or less. The cell container sheet according to claim 1 .

5. The cell container sheet has an oxygen permeability of 7500 cm at 23° C. ３ / (m ２ ・24h・ATM) or more 150,000cm ３ / (m ２ ・24h・atm) or less, The cell container sheet according to claim 1 .

6. The cell container sheet has a water vapor permeability of 10 g / (m ２ ・24h) or more 100g / (m ２ - 24h) or less, The cell container sheet according to claim 1 .

7. Sterility assurance level (SAL) of medical devices measured in accordance with BS EN556-1:2001 is 10 －６ Below is the The cell container sheet according to claim 1 .

8. When the cell container sheet is attached to a well plate, the thickness of the sheet at the periphery of the well is thinner than the thickness of the sheet at the center of the well. The cell container sheet according to claim 1 .

9. a multi-layer sheet having an elastomer layer containing one or more thermoplastic elastomers selected from the group consisting of polyolefin-based elastomers, styrene-based elastomers, polyurethane-based elastomers, polyester-based elastomers, and polyamide-based elastomers; The cell container sheet according to claim 1 .

10. The cell container sheet has a maximum value of loss tangent (tan δ) of dynamic viscoelasticity in the range of 10° C. or more and 100° C. or less and in the range of −70° C. or more and −20° C. or less, the maximum value being determined by dynamic viscoelasticity measurement performed under conditions of a temperature rise rate of 4° C. / min, a frequency of 1.59 Hz, and a distortion amount of 0.1%, and the maximum value of the loss tangent in the range of 10° C. or more and 100° C. or less is 0.5 or more and 3.5 or less; The maximum value of the loss tangent in the range of -70 ° C. or more and -20 ° C. or less is 0.05 or more and 1.0 or less. The cell container sheet according to claim 9 .

11. The ratio of the thickness of the adhesive layer to the thickness of the elastomer layer (adhesive layer / elastomer layer) is 0.9 or more and 5.0 or less. The cell container sheet according to claim 9 .

12. The elastomer layer is laminated at least partially in contact with the adhesive layer. The cell container sheet according to claim 9 .

13. The cell container sheet has a tab on its periphery for peeling it from the cell container. The cell container sheet according to claim 1 .

14. The tab is disposed at a position that protrudes from the peripheral portion of another member when the cell container sheet is attached to the other member of the cell container. The cell container sheet according to claim 13.

15. The cell container has an indication portion that indicates a surface to be brought into contact with another member when the cell container is brought into close contact with the other member. The cell container sheet according to claim 1 .

16. A lid material for cell containers. The cell container sheet according to claim 1 .

17. The base material of the cell container, The cell container sheet according to claim 1 .

18. The cell container sheet according to any one of claims 1 to 17, A substrate having a through hole or a recess, The cell container sheet is in close contact with the upper or bottom surface of the through-hole, or the upper surface of the recess. cell container.

19. the substrate having a plurality of wells; The cell container sheet has a thickness smaller at the periphery of the well than at the center of the well. The cell container of claim 18.

20. A step of forming a cell container by adhering the cell container sheet according to any one of claims 1 to 17 to a substrate having a through-hole or a recess; Cultivating cells in the cell container; removing the cell container sheet from the cell container; A method for culturing cells comprising the steps of:

Images