Freezing preservation container

Abstract

To provide a freezing preservation container which can freeze and thaw a biological sample.SOLUTION: A freezing preservation container 1 comprises: a bottom sheet-like member 11; a lid sheet-like member 14 laminated on the bottom sheet-like member 11; a three-dimensional storage part 105 which is formed of the bottom sheet-like member 11 and the lid sheet-like member 14 at an inside in a plane direction of the bottom sheet-like member 11 and the lid sheet-like member 14, is molded into a three-dimensional state with a thickness in a thickness direction of the bottom sheet-like member 11 and the lid sheet-like member 14, and stores therein a biological sample 101; and a biological sample holding part 21 for holding the biological sample 101 stored in the three-dimensional storage part 105 in a center in the thickness direction of the three-dimensional storage part 105.SELECTED DRAWING: Figure 5

Description

[Technical field]

[0001] The present invention relates to a cryopreservation container for preserving a biological sample. [Background technology]

[0002] Conventionally, a cryopreservation container is known in which an opening communicating with a container body is sealed by thermal welding (see, for example, Patent Document 1). Biological samples such as blood vessels and other cells are contained in the cryopreservation container, and the cryopreservation container is frozen and preserved. The cryopreservation container described in Patent Document 1 has an outer surface formed in an elliptical shape in cross section. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2007-302567 A Summary of the Invention [Problem to be solved by the invention]

[0004] The cryopreservation container described in Patent Document 1 has an outer surface formed in an elliptical shape in cross section, and the thickness is not constant. If the thickness of the cryopreservation container is not constant, the distance from the outer surface of the cryopreservation container to the biological sample, such as blood vessels or other cells, contained inside the cryopreservation container is not constant, and the cryopreservation container may not be frozen uniformly when frozen, or may not be thawed uniformly when thawed. This causes variation in the state of the biological sample, making it difficult to maintain a constant quality of the biological sample.

[0005] An object of the present invention is to provide a cryopreservation container that allows biological samples to be frozen and thawed uniformly. [Means for solving the problem]

[0006] The present invention relates to a cryopreservation container comprising a bottom sheet-like member, a lid sheet-like member laminated on the bottom sheet-like member, a three-dimensional storage section formed by the bottom sheet-like member and the lid sheet-like member on the inside of the planar direction of the bottom sheet-like member and the lid sheet-like member, having a thickness in the thickness direction of the bottom sheet-like member and the lid sheet-like member, and storing a biological sample therein, and a biological sample holding section that holds the biological sample stored in the three-dimensional storage section at the center of the thickness direction of the three-dimensional storage section.

[0007] It is also preferable that the biological sample holder holds the biological sample by sandwiching it in the thickness direction of the bottom sheet-like member and the lid sheet-like member.

[0008] It is also preferable that the biological sample is formed in a sheet-like form, and the biological sample holding section has two mesh sheet sections that hold the sheet-like biological sample by sandwiching it in the thickness direction between the bottom sheet-like member and the lid sheet-like member.

[0009] In addition, it is preferable that the two mesh sheet portions hold the biological sample inside the three-dimensional storage section which is arranged inside the planar direction of the bottom sheet member and the lid sheet member, and that the planar outer peripheral edges of the bottom sheet member and the lid sheet member are held by being sandwiched between the bottom sheet member and the lid sheet member.

[0010] It is also preferable that the bottom side sheet-like member has a first three-dimensional shape portion formed in a concave shape with the lid side sheet-like member side open, and a first three-dimensional shape portion surrounding sheet-like member portion arranged around the first three-dimensional shape portion, and the lid side sheet-like member has a second three-dimensional shape portion formed in a concave shape with the bottom side sheet-like member side open, and a second three-dimensional shape portion surrounding sheet-like member portion arranged around the second three-dimensional shape portion, and the three-dimensional storage portion is configured such that the open portions of the first three-dimensional shape portion and the second three-dimensional shape portion are arranged opposite each other.

[0011] Furthermore, it is preferable that the two mesh sheet portions are composed of a first mesh sheet portion having a first concave portion that is concave in a stepped manner on one side in the thickness direction of the bottom sheet member and the lid sheet member, and a second mesh sheet portion having a second concave portion that is concave in a stepped manner on one side in the same thickness direction as the first concave portion is concave in the thickness direction of the bottom sheet member and the lid sheet member, and that the first concave portion and the second concave portion are stacked and arranged with the biological sample sandwiched between the bottom of the first concave portion and the bottom of the second concave portion. Effect of the Invention

[0012] According to the present invention, it is possible to provide a cryopreservation container capable of uniformly freezing and thawing a biological sample. [Brief description of the drawings]

[0013] [Figure 1] 1 is a perspective view of a cryopreservation container according to a first embodiment of the present invention, viewed obliquely from above. [Diagram 2] FIG. 2 is a perspective view of the cryopreservation container according to the first embodiment, seen obliquely from below. [Diagram 3] FIG. 2 is a plan view of the cryopreservation container according to the first embodiment. [Figure 4] FIG. 2 is an exploded perspective view of the cryopreservation container according to the first embodiment. [Diagram 5] FIG. 2 is a schematic cross-sectional view taken along line AA in FIG. [Figure 6] FIG. 2 is a schematic cross-sectional view taken along line BB in FIG. [Figure 7] FIG. 2 is a plan view of the cryopreservation container according to the first embodiment, showing the positions of each sealing portion. [Figure 8] 3A to 3C are cross-sectional views showing a method of using the cryopreservation container according to the first embodiment. [Figure 9] FIG. 2 is a plan view showing a method of using the cryopreservation container according to the first embodiment. [Figure 10] 1 is a perspective view of a cryopreservation container according to a second embodiment of the present invention, seen obliquely from above. FIG. [Figure 11]FIG. 11 is a perspective view of a cryopreservation container according to a second embodiment, as viewed obliquely from below. [Figure 12] FIG. 11 is a plan view of a cryopreservation container according to a second embodiment. [Figure 13] FIG. 11 is an exploded perspective view of a cryopreservation container according to a second embodiment. [Figure 14] 11 is a schematic cross-sectional view taken along line CC in FIG. 10. FIG. [Figure 15] FIG. 11 is a schematic cross-sectional view taken along line DD in FIG. [Figure 16] FIG. 11 is a plan view of a cryopreservation container according to a second embodiment, showing the positions of each sealing portion. [Figure 17] 6A to 6C are cross-sectional views showing a method of using the cryopreservation container according to the second embodiment. [Figure 18] 11 is a plan view showing a method of using the cryopreservation container according to the second embodiment. FIG. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] Hereinafter, preferred embodiments of the cryopreservation container 1 as a medical container of the present invention will be described with reference to the drawings. The cryopreservation container 1 of the present invention is used in medical treatment to cryopreserve biological samples such as cell sheets and other cells.

[0015] A cryopreservation container 1 of the first embodiment will be described with reference to Figs. 1 to 7. In the description of this embodiment, the direction in which one side of the container body 10 formed in a quadrangular shape in a plan view of the cryopreservation container 1 extends is referred to as the X direction, and the direction in which the other side of the container body 10 extends is referred to as the Y direction. The X direction and the Y direction are perpendicular to each other. The stacking direction of the multiple sheet-like members 11, 12, 13, and 14 of the container body 10 is referred to as the Z direction.

[0016] As shown in Figs. 1 to 4, the cryopreservation container 1 of the first embodiment includes a container body 10 and a liquid introduction tube 20 (tube) capable of introducing a cryoprotective liquid 102 (protective agent) (see Fig. 5). The container body 10 is configured by stacking a plurality of sheet-like members 11, 12, 13, and 14. As shown in Figs. 5 and 6, in the container body 10, a three-dimensional container section 105 (described later) formed by a first sheet-like member 11 (bottom-side sheet-like member) and a third sheet-like member 14 (lid-side sheet-like member) contains a cell sheet 101 (biological sample) in a state in which the cell sheet 101 is held by a cell sheet holding structure 21 (biological sample holding section). The cell sheet 101 is formed in a sheet shape by culturing cells collected from human tissue. The cryopreservation container 1 is cryopreserved in a state in which a biological sample such as the cell sheet 101 is contained.

[0017] As shown in Figures 3 to 7, the container body 10 comprises a first sheet-like member 11 (bottom sheet-like member), a heat-resistant sheet-like member 12, a second sheet-like member 13, a third sheet-like member 14 (lid-side sheet-like member), a weak seal portion 15 (first seal portion), a strong seal portion 16 (third seal portion), a sealing seal portion 17 (second seal portion), a liquid introduction tube side sealing seal portion 18, and a cell sheet holding structure 21 (biological sample holding portion).

[0018] The first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14 are made of a flexible thermoplastic resin such as EVA resin (ethylene-vinyl acetate copolymer resin). The heat-resistant sheet-like member 12 is made of a material sheet made of a heat-resistant fluorine material, preferably a sheet made of a material such as PTFE (polytetrafluoroethylene). The first sheet-like member 11, the heat-resistant sheet-like member 12, the second sheet-like member 13, and the third sheet-like member 14 are made of a freeze-resistant material so that they can be frozen.

[0019] As shown in FIG. 4, the container body 10 is configured by stacking a first sheet-like member 11, a heat-resistant sheet-like member 12, a second sheet-like member 13 and a third sheet-like member 14, with a cell sheet 101 held by a cell sheet holding structure 21 disposed between the second sheet-like member 13 and the third sheet-like member 14, and any two of the sheet-like members being partially joined together at the weak seal portion 15, the strong seal portion 16, the sealing seal portion 17 and the liquid introduction tube side sealing portion 18, as shown in FIGS. 5 to 7.

[0020] 4, the first sheet-like member 11, the heat-resistant sheet-like member 12, the second sheet-like member 13, and the third sheet-like member 14 are stacked in this order from one side Z1 to the other side Z2 in the Z direction. The cell sheet holding structure 21 has two sheet-like mesh sheet members 211, 212 (mesh sheet portion) stacked in the Z direction. The cell sheet 101 is placed between the second sheet-like member 13 and the third sheet-like member 14 in a state where it is sandwiched and held between the two mesh sheet members 211, 212.

[0021] When the container body 10 is in a state in which the first sheet-like member 11, the heat-resistant sheet-like member 12, the second sheet-like member 13 and the third sheet-like member 14 are stacked, as shown in Figure 3, the container body 10 has an approximately rectangular outer shape in a plan view, with one side extending in the X direction and the other side extending in the Y direction.

[0022] The liquid introduction tube 20 is attached to one end of the container body 10 in the Y direction. The liquid introduction tube 20 is disposed between the first sheet-like member 11 and the third sheet-like member 14 on one side of the cryopreservation container 1. The liquid introduction tube 20 is capable of introducing a cryoprotective liquid 102 (see FIG. 5 ) into the container body 10. The liquid introduction tube 20 may be configured to be disposed between the first sheet-like member 11 and the second sheet-like member 13 on one side of the cryopreservation container 1, or may be configured to be disposed between the second sheet-like member 13 and the third sheet-like member 14.

[0023] 4, the first sheet-like member 11 has a bottom-side peripheral sheet-like portion 111 (first three-dimensional shaped portion peripheral sheet-like portion) and a bottom-side three-dimensional shaped portion 112 (first three-dimensional shaped portion). The bottom-side peripheral sheet-like portion 111 has a substantially rectangular outer shape in a plan view, with one side extending in the X direction and the other side extending in the Y direction. The bottom-side peripheral sheet-like portion 111 is disposed around the bottom-side three-dimensional shaped portion 112.

[0024] The bottom side three-dimensional shape portion 112 is formed by three-dimensional molding as shown in Figures 4 to 6. The bottom side three-dimensional shape portion 112 is arranged inside the planar direction of the bottom side peripheral sheet-like portion 111 in a planar view. The bottom side three-dimensional shape portion 112 is formed into a three-dimensional shape with a concave shape that is recessed from the bottom side peripheral sheet-like portion 111 to one side Z1 in the Z direction with the third sheet-like member 14 side open. The bottom side three-dimensional shape portion 112 is formed recessed by a predetermined thickness from the inner peripheral edge of the bottom side peripheral sheet-like portion 111 to one side Z1 in the Z direction, and is formed into a three-dimensional shape like a circular short cylinder that has a bottom surface at the end of one side Z1 in the Z direction and is open to the other side Z2.

[0025] The heat-resistant sheet member 12 is arranged in a stacked manner on the other side Z2 of the first sheet member 11 in the Z direction. The heat-resistant sheet member 12 is formed in a U-shape with a predetermined width that opens on the liquid introduction tube 20 side at a position away from the bottom side three-dimensional shape portion 112 of the first sheet member 11 in the X direction and Y direction in a plan view. The heat-resistant sheet member 12 has an opening hole 121 (heat-resistant sheet member opening hole) in a portion corresponding to the bottom side three-dimensional shape portion 112. The opening hole 121 of the heat-resistant sheet member 12 is open only on one side where the liquid introduction tube 20 of the cryopreservation container 1 is sealed. The opening hole 121 opens on the liquid introduction tube 20 side in the Y direction so as to avoid the bottom side three-dimensional shape portion 112 and open in a range wider than the bottom side three-dimensional shape portion 112.

[0026] 5 to 7, the heat-resistant sheet member 12 is disposed between the first sheet member 11 and the second sheet member 13 (described later) in a position where the weak seal portion 15 (described later) and the strong seal portion 16 (described later) are not formed in a plan view and in the outer circumferential direction of the bottom three-dimensional shape portion 112 avoiding the weak seal portion 15 and the strong seal portion 16. The predetermined width of the heat-resistant sheet member 12 is preferably the same as that of the sealing portion 17 (described later) because the sealing portion 17 is sealed in the portion laminated on the heat-resistant sheet member 12, and is, for example, preferably 2 to 20 mm, more preferably 10 to 15 mm.

[0027] 4, the second sheet-like member 13 is arranged on the other side Z2 of the first sheet-like member 11 in the Z direction, stacked with the heat-resistant sheet-like member 12 in between. That is, the second sheet-like member 13 is arranged on the other side Z2 of the heat-resistant sheet-like member 12 in the Z direction. The outer shape of the second sheet-like member 13 is formed in a planar shape of the same size as the outer shape of the first sheet-like member 11 in a plan view, with one side extending in the X direction and the other side extending in the Y direction.

[0028] The second sheet-like member 13 is formed in a U-shape in a plan view such that the liquid introduction tube 20 side is open at a position avoiding the bottom side three-dimensional shape portion 112 of the first sheet-like member 11. The second sheet-like member 13 has an opening hole 131 (second sheet-like member opening hole) at a portion corresponding to the bottom side three-dimensional shape portion 112. The opening hole 131 is shaped to correspond to the bottom side three-dimensional shape portion 112 so as to avoid the bottom side three-dimensional shape portion 112, and is open on the liquid introduction tube 20 side in the Y direction.

[0029] The third sheet-like member 14 is arranged in a stacked manner in the Z direction on the other side Z2 of the second sheet-like member 13 (the side of the second sheet-like member 13 opposite the first sheet-like member 11) with the two mesh sheet members 211, 212 of the cell sheet holding structure 21 sandwiched between the second sheet-like member 13. The third sheet-like member 14 is formed in a planar shape having the same size as the outer shape of the first sheet-like member 11 in a plan view, and is formed so as to cover the bottom side three-dimensional shape portion 112 of the first sheet-like member 11.

[0030] 4, the third sheet-like member 14 has a lid-side peripheral sheet portion 141 (second three-dimensional shaped portion peripheral sheet portion) and a lid-side three-dimensional shaped portion 142 (second three-dimensional shaped portion). The lid-side peripheral sheet portion 141 has a substantially rectangular shape in plan view, with one side extending in the X direction and the other side extending in the Y direction. The lid-side peripheral sheet portion 141 is formed in a planar shape having the same size as the outer shape of the bottom-side peripheral sheet portion 111 of the first sheet-like member 11 in plan view. The lid-side peripheral sheet portion 141 is disposed around the lid-side three-dimensional shaped portion 142.

[0031] The lid-side three-dimensional shape portion 142 is formed by three-dimensional molding as shown in Figs. 4 to 6. The lid-side three-dimensional shape portion 142 is formed in a circular shape in a plan view, and is disposed inside the planar direction of the lid-side peripheral sheet-like portion 141. The lid-side three-dimensional shape portion 142 is formed in a concave three-dimensional shape that opens on the first sheet-like member 11 side and is concave from the lid-side peripheral sheet-like portion 141 to the other side Z2 in the Z direction. The lid-side three-dimensional shape portion 142 is formed by concaving a predetermined thickness from the inner peripheral edge of the lid-side peripheral sheet-like portion 141 to the other side Z2 in the Z direction, and is formed in a three-dimensional shape of a circular short cylinder having a top surface at the end of the other side Z2 in the Z direction and one side Z1 open. In this embodiment, the thickness in the Z direction of the lid-side three-dimensional shape portion 142 is formed to be the same thickness as the thickness in the Z direction of the bottom-side three-dimensional shape portion 112 of the first sheet-like member 11.

[0032] The third sheet-like member 14 is arranged by being stacked on the other side Z2 in the Z direction of the first sheet-like member 11, with the opening of the bottom side three-dimensional shape portion 112 of the first sheet-like member 11 facing one side Z1 in the Z direction of the opening of the lid side three-dimensional shape portion 142 of the third sheet-like member 14. With the open openings of the bottom side three-dimensional shape portion 112 of the first sheet-like member 11 and the lid side three-dimensional shape portion 142 of the third sheet-like member 14 arranged facing each other, the bottom side three-dimensional shape portion 112 of the first sheet-like member 11 and the lid side three-dimensional shape portion 142 of the third sheet-like member 14 are stacked in the Z direction to form a three-dimensional containing section 105 having a thickness in the Z direction.

[0033] The three-dimensional storage section 105 is configured by arranging the open openings of the bottom side three-dimensional shape portion 112 and the lid side three-dimensional shape portion 142 facing each other. The three-dimensional storage section 105 is formed by the bottom side three-dimensional shape portion 112 of the first sheet-like member 11 and the lid side three-dimensional shape portion 142 of the third sheet-like member 14 on the inner side in the planar direction of the first sheet-like member 11 and the third sheet-like member 14, and is three-dimensionally formed with a thickness in the thickness direction of the first sheet-like member 11 and the third sheet-like member 14 so as to protrude on both sides Z1 and Z2 in the Z direction of the container body 10 of the cryopreservation container 1.

[0034] The three-dimensional containing section 105 contains biological samples such as the cell sheet 101 and other cells in a state held by the cell sheet holding structure 21. The three-dimensional containing section 105 has an outer shape formed with a thickness in the Z direction, and has a circular outer shape in a plan view. As shown in Fig. 5, the three-dimensional containing section 105 has a rectangular cross-sectional shape extending in the Y direction when cut along the X direction, and as shown in Fig. 6, the three-dimensional containing section 105 has a rectangular cross-sectional shape extending in the Y direction when cut along the Y direction.

[0035] The cell sheet holding structure 21 has two mesh sheet members 211, 212 (mesh sheet members). Both of the two mesh sheet members 211, 212 are formed in a mesh-like, planar sheet shape. The two mesh sheet members 211, 212 hold the cell sheet 101 held in the three-dimensional containing section 105 by sandwiching it in the thickness direction of the first sheet-like member 11 and the third sheet-like member 14. The cell sheet 101 held by the two mesh sheet members 211, 212 of the cell sheet holding structure 21 is held in the three-dimensional containing section 105.

[0036] The two mesh-like sheet members 211, 212 are stacked in this order from one side Z1 to the other side Z2 in the Z direction with the cell sheet 101 sandwiched and held in the Z direction between the second sheet-like member 13 and the third sheet-like member 14. The cell sheet 101 sandwiched between the two mesh-like sheet members 211, 212 is placed inside the three-dimensional containing section 105 formed by the first sheet-like member 11 and the third sheet-like member 14.

[0037] Here, since the Z-direction thickness of the lid-side three-dimensional shape portion 142 of the third sheet-like member 14 constituting the three-dimensional containing section 105 and the Z-direction thickness of the bottom-side three-dimensional shape portion 112 of the first sheet-like member 11 are formed to be the same thickness, the cell sheet 101 held by the two mesh-like sheet members 211, 212 is positioned at the center in the Z direction of the three-dimensional containing section 105. As a result, the two mesh-like sheet members 211, 212 hold the cell sheet 101 at the center in the Z direction, which is the thickness direction of the three-dimensional containing section 105.

[0038] The two mesh-like sheet members 211, 212 are formed in a planar shape that is larger than the opening area of ​​the bottom-side three-dimensional shape portion 112 and the lid-side three-dimensional shape portion 142 that constitute the three-dimensional containing section 105, and smaller than the outer size of the second sheet-like member 13 and the third sheet-like member 14. The two mesh-like sheet members 211, 212 hold the cell sheet 101 inside the three-dimensional containing section 105. The outer peripheral edges of the two mesh-like sheet members 211, 212 are sandwiched and held between the second sheet-like member 13 and the third sheet-like member 14.

[0039] A liquid introduction tube 20 disposed at one end of the container body 10 in the Y direction communicates with the three-dimensional containing section 105 on one side in the Y direction. In the three-dimensional containing section 105, a cryoprotective liquid 102 (see FIG. 5 ) is introduced by the liquid introduction tube 20 while the cell sheet 101 is contained therein. In this embodiment, the cell sheet 101 is formed in a disk shape having a diameter smaller than the inner diameter of the three-dimensional containing section 105 (the bottom side three-dimensional shape section 112 and the lid side three-dimensional shape section 142) so as to be contained in the three-dimensional containing section 105 (the bottom side three-dimensional shape section 112 and the lid side three-dimensional shape section 142). As a result, when the cryopreservation container 1 is frozen, the three-dimensional containing section 105 is filled with the cryoprotective liquid 102 while the cell sheet 101 is contained therein, as shown in FIG. 5 .

[0040] As shown in Fig. 5 to Fig. 7, the weak seal portion 15, the strong seal portion 16, the sealing seal portion 17, and the liquid introduction tube side sealing portion 18 are sealing portions formed by joining any two of the first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14 by heat, such as heat sealing or ultrasonic sealing. Note that the weak seal portion 15, the strong seal portion 16, the sealing seal portion 17, and the liquid introduction tube side sealing portion 18 are not limited to sealing portions joined by heat. For example, they may be sealing portions joined by high-frequency welding or ultrasonic welding.

[0041] 7, the weak seal portion 15 is formed on the outer periphery of the three-dimensional storage portion 105 at the outer edges of the bottom side three-dimensional shape portion 112 of the first sheet-like member 11 and the lid side three-dimensional shape portion 142 of the third sheet-like member 14 in a plan view. The weak seal portion 15 seals three sides of the cryopreservation container 1 where the liquid introduction tube 20 is not disposed.

[0042] The weak seal portion 15 is formed in a U-shape in plan view with the liquid introduction tube 20 side open, and seals the first sheet-like member 11 and the second sheet-like member 13 so as to surround the outer periphery of the bottom side three-dimensional shape portion 112 of the first sheet-like member 11, as shown in Fig. 5. The weak seal portion 15 is a seal portion where the first sheet-like member 11 and the second sheet-like member 13 are sealed by heat with a seal strength that allows them to be opened by hand. The weak seal portion 15 has a seal strength that prevents the cryoprotectant liquid 102 injected into the three-dimensional storage portion 105 from leaking out to the outside.

[0043] As shown in Fig. 7, the strong seal portion 16 is formed on the outer periphery of the first sheet-like member 11 outside the weak seal portion 15 with respect to the bottom three-dimensional shape portion 112 in a plan view. The strong seal portion 16 seals three sides of the cryopreservation container 1 where the liquid introduction tube 20 is not disposed on the outer periphery side of the weak seal portion 15. The strong seal portion 16 seals the first sheet-like member 11 and the second sheet-like member 13 at a position away from the outer edge of the U-shaped portion of the weak seal portion 15 along the outer edge of the first sheet-like member 11 and the outer edge of the second sheet-like member 13, as shown in Fig. 5, so as to surround the U-shape with a predetermined width that opens on the liquid introduction tube 20 side.

[0044] The strong seal portion 16 seals between the first sheet-like member 11 and the second sheet-like member 13 so as to surround the periphery of the weak seal portion 15 and the heat-resistant sheet-like member 12. The strong seal portion 16 is a seal portion in which the first sheet-like member 11 and the second sheet-like member 13 are sealed by heat with a seal strength stronger than that of the weak seal portion 15. The strong seal portion 16 is a seal portion that is sealed so that the surrounding liquid nitrogen does not come into contact with the weak seal portion 15 when frozen and the heat-resistant sheet-like member 12 does not come into contact with liquid nitrogen, and has a seal strength that makes it difficult for a person to open it by hand.

[0045] As shown in Fig. 7, the sealing portion 17 is formed between the weak seal portion 15 and the strong seal portion 16 in plan view, and seals in a U-shape with a predetermined width that opens to the liquid introduction tube 20 side. The sealing portion 17 is formed at at least a part of the position where the heat-resistant sheet member 12 is disposed in plan view. The predetermined width of the sealing portion 17 is preferably, for example, 2 to 20 mm, and more preferably 10 to 15 mm. This prevents the sealing portion 17 from shifting and overlapping with other seal portions, and ensures reliable sealing at the sealing position.

[0046] As shown in FIG. 5, the sealing portion 17 is a sealing portion that seals the portion where the second sheet-like member 13 and the third sheet-like member 14 that are not sealed by the weak sealing portion 15 and the strong sealing portion 16 are laminated on the heat-resistant sheet-like member 12 so that the cryoprotective liquid 102 injected into the three-dimensional storage portion 105 (the bottom side three-dimensional shape portion 112 and the lid side three-dimensional shape portion 142) does not leak out to the outside. In addition, when sealing the second sheet-like member 13 and the third sheet-like member 14, the sealing portion 17 seals the peripheral portions of the two mesh-like sheet members 211 and 212 of the cell sheet holding structure 21 together with the second sheet-like member 13 and the third sheet-like member 14. The sealing portion 17 has a seal strength that is difficult for a person to open by hand, and is a seal portion that is sealed by heat with a seal strength stronger than that of the weak sealing portion 15. The seal strength of the sealing portion 17 may be the same as or different from the seal strength of the strong sealing portion 16.

[0047] 7, the liquid introduction tube side sealing part 18 seals the first sheet-like member 11 and the third sheet-like member 14 with the liquid introduction tube 20 disposed between them at the end on the liquid introduction tube 20 side in the Y direction. The liquid introduction tube side sealing part 18 seals the first sheet-like member 11 and the third sheet-like member 14 only on one side of the cryopreservation container 1 that includes the liquid introduction tube 20 with a stronger seal strength than the weak seal part 15. The liquid introduction tube side sealing part 18 is a sealing part that has a seal strength that makes it difficult for a person to open it by hand.

[0048] The liquid introduction tube 20 introduces liquid into the three-dimensional storage section 105. The liquid introduction tube 20 is made of a thermoplastic resin such as EVA resin. In the first embodiment, the liquid introduction tube 20 is disposed between the first sheet-like member 11 and the second sheet-like member 13 so that one end side of the liquid introduction tube 20 communicates with the three-dimensional storage section 105 (the bottom side three-dimensional shaped portion 112 and the lid side three-dimensional shaped portion 142) of the first sheet-like member 11. The other end side of the liquid introduction tube 20 is aseptically and airtightly connected to a liquid storage container (not shown) that contains, for example, the cryoprotectant liquid 102.

[0049] In the cryopreservation container 1 configured as described above, the cell sheet 101 is sandwiched between two mesh-like sheet members 211, 212 and disposed at the center position in the Z direction in the three-dimensional containing section 105, so that the cell sheet 101 can be disposed at the center in the Z direction. This allows the distance from the cell sheet 101 disposed inside the three-dimensional containing section 105 to the outer surface of one side Z1 and the outer surface of the other side Z2 in the Z direction of the three-dimensional containing section 105 to be the same. Therefore, the cell sheet 101 contained in the cryopreservation container 1 can be frozen and thawed uniformly.

[0050] As shown in Figs. 1 to 3, the first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14 have marks 115, 116 that serve as cutting guides when removing the contents. In this embodiment, the mark 115 is a mark for cutting the first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14 that are arranged in a stacked manner along the X direction. The mark 116 is a mark for cutting the first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14 that are arranged in a stacked manner along the Y direction. As shown in Fig. 4, the marks 115, 116 are formed by overlapping a pair of recessed portions 11d, 11e of the first sheet-like member 11, a pair of recessed portions 13d, 13e of the second sheet-like member 13, and a pair of recessed portions 14d, 14e of the third sheet-like member 14.

[0051] In this way, by providing the mark 115 serving as a guide for cutting along the X direction and the mark 116 serving as a guide for cutting along the Y direction, it is possible to cut along the X direction as well as along the Y direction. When cutting the first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14, it is possible to cut along the Y direction after using the mark 115 to cut along the X direction, or to cut along the Y direction after using the mark 116 to cut along the X direction.

[0052] Next, a method for storing a biological sample such as a cell sheet 101 in the cryopreservation container 1 will be described. Referring to FIG. 4, the cryopreservation container 1 is produced in a factory by arranging a first sheet-like member 11, a heat-resistant sheet-like member 12, and a second sheet-like member 13 in a layered manner, and sealing them at the weak seal portion 15 and the strong seal portion 16 as shown in FIGS. 5 to 7. When the cryopreservation container 1 is shipped from the factory, the third sheet-like member 14 is shipped in a state in which only the liquid introduction tube side sealing portion 18 is sealed. Furthermore, when shipped from the factory, the two mesh-like sheet members 211, 212 of the cell sheet holding structure 21 and the cell sheet 101 are not arranged between the second sheet-like member 13 and the third sheet-like member 14.

[0053] After shipment from the factory, the cryopreservation container 1 is in a state in which the first sheet-like member 11, the heat-resistant sheet-like member 12 and the second sheet-like member 13 are stacked and arranged as shown in Figures 4 to 6, and sealed at the weak seal portion 15 and the strong seal portion 16.The user then places two mesh-like sheet members 211, 212 sandwiching a cell sheet 101 between the second sheet-like member 13 and the third sheet-like member 14, as shown in Figures 5 and 6.

[0054] Next, as shown in Figures 5 and 6, the third sheet-like member 14 is placed on the other side Z2 of the two mesh-like sheet members 211, 212 arranged on the other side Z2 of the second sheet-like member 13, and the second sheet-like member 13 and the third sheet-like member 14 are sealed by the sealing portion 17 at the portion laminated on the heat-resistant sheet-like member 12.

[0055] In this case, since the heat-resistant sheet-like member 12 is disposed on one side Z1 of the second sheet-like member 13 and the third sheet-like member 14, the first sheet-like member 11 does not melt due to heat, and the second sheet-like member 13 and the third sheet-like member 14 are sealed together by the sealing portion 17. Furthermore, when the sealing portion 17 seals the weak seal portion 15 and the strong seal portion 16, the peripheral portions of the two mesh-like sheet members 211, 212 of the cell sheet holding structure 21 disposed in the heat-resistant sheet-like member 12 from the inside to the middle of the width direction of the heat-resistant sheet-like member 12 are simultaneously sealed.

[0056] 7, a cryoprotectant 102 is introduced into the three-dimensional container 105, and the opening of the liquid introduction tube 20 is sealed. In this way, the user can contain and seal a biological sample, such as a cell sheet 101 or other cells, in the cryopreservation container 1. The cryopreservation container 1 containing the cell sheet, other cells, etc. is then frozen and preserved.

[0057] Here, the cell sheet 101 held by the two mesh-like sheet members 211, 212 is disposed inside the three-dimensional containing section 105 at the center in the Z direction of the three-dimensional containing section 105. As a result, the two mesh-like sheet members 211, 212 hold the cell sheet 101 at the center in the Z direction, which is the thickness direction of the three-dimensional containing section 105. Therefore, the distance from the cell sheet 101 disposed inside the three-dimensional containing section 105 to the outer surface of one side Z1 and the outer surface of the other side Z2 in the Z direction of the three-dimensional containing section 105 can be set to the same distance, so that the cell sheet 101 contained in the cryopreservation container 1 can be frozen uniformly.

[0058] Next, a method for opening the cryopreservation container 1 of the first embodiment will be described. When opening the cryopreservation container 1 to take out the cell sheet 101 and other biological samples such as cells stored in the cryopreservation container 1, the frozen cryopreservation container 1 is first thawed. Here, the two mesh-like sheet members 211, 212 hold the cell sheet 101 at the center in the Z direction, which is the thickness direction of the three-dimensional storage section 105. Therefore, the distance from the cell sheet 101 placed inside the three-dimensional storage section 105 to the outer surface of one side Z1 and the outer surface of the other side Z2 in the Z direction of the three-dimensional storage section 105 can be set to the same distance, so that the cell sheet 101 stored in the cryopreservation container 1 can be thawed uniformly.

[0059] Next, as shown in the upper and middle diagrams of Figure 8 and Figure 9, the outer peripheral edge portion 19 (outside the end portion 11a of the first sheet-like member 11, the end portion 13a of the second sheet-like member 13, the end portion 14a of the third sheet-like member 14, and the sealing portion 17) including the portion where the strong sealing portion 16 of the container body 10 is formed is cut off using a cutting tool such as scissors.

[0060] Specifically, as shown in Fig. 9, the first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14 are cut along the X direction from the mark 115, so that the strong seal portion 16 extending in the X direction formed at the end of the cryopreservation container 1 on the side opposite to the side where the liquid introduction tube 20 is arranged in the Y direction can be cut off. Then, at both ends in the X direction, the first sheet-like member 11, the second sheet-like member 13, and the third sheet-like member 14 are cut along the Y direction on the inside in the X direction of the strong seal portion 16 extending in the Y direction, so that the strong seal portion 16 extending in the Y direction formed at both ends in the X direction of the cryopreservation container 1 can be cut off. Then, as shown in the middle diagram of Fig. 8, the heat-resistant sheet member 12 arranged between the first sheet-like member 11 and the second sheet-like member 13 is taken out.

[0061] 8, the first sheet-like member 11 and the second sheet-like member 13 are pulled at their outer edges toward one side Z1 and the other side Z2 in the Z direction so that they are separated from each other, and the first sheet-like member 11 and the second sheet-like member 13 are peeled off at the weak seal portion 15 to open the container body 10. The weak seal portion 15 has a seal strength that prevents the cryoprotectant liquid 102 injected into the three-dimensional storage portion 105 from leaking out, has a seal strength weaker than that of the sealing seal portion 17, and is a seal portion that is heat-sealed with a seal strength that can be opened by hand, and the second sheet-like member 13 is strongly sealed to the third sheet-like member 14 side, so that the first sheet-like member 11 and the second sheet-like member 13 can be easily peeled off at the weak seal portion 15. After opening at the weak seal portion 15, the two mesh sheet members 211 and 212 are partially torn to remove the cell sheet 101.

[0062] This allows the container body 10 to be opened and the cell sheet 101 contained in the three-dimensional containing portion 105 to be removed. Before cutting the outer periphery 19 including the strong seal portion 16, the outer periphery is sealed by the strong seal portion 16, and after removing the strong seal portion 16, the first sheet-like member 11 and the second sheet-like member 13 can be peeled away from each other to open the container at the weak seal portion 15. This prevents the cell sheet 101 contained in the three-dimensional containing portion 105 from being contaminated.

[0063] The cryopreservation container 1 of the first embodiment described above provides the following advantages. (1) The cryopreservation container 1 is configured to include a first sheet-like member 11, a third sheet-like member 14 laminated on the first sheet-like member 11, a three-dimensional storage section 105 formed by the first sheet-like member 11 and the third sheet-like member 14 on the inner side in the planar direction of the first sheet-like member 11 and the third sheet-like member 14, having a thickness in the thickness direction of the first sheet-like member 11 and the third sheet-like member 14 and storing a cell sheet 101 therein, and a cell sheet retaining structure 21 that retains the cell sheet 101 stored in the three-dimensional storage section 105 at the center in the thickness direction of the three-dimensional storage section 105.

[0064] This allows the distance from the cell sheet 101 placed inside the three-dimensional storage section 105 to the outer surface of one side Z1 and the outer surface of the other side Z2 in the Z direction of the three-dimensional storage section 105 to be the same. Therefore, when the cryopreservation container 1 is frozen, the cell sheet 101 and the cryoprotective solution 102 placed inside the three-dimensional storage section 105 can be frozen uniformly, and when the cryopreservation container 1 is thawed, the cell sheet 101 and the cryoprotective solution 102 placed inside the three-dimensional storage section 105 can be thawed uniformly.

[0065] (2) The cell sheet holding structure 21 is configured to hold the cell sheet 101 by sandwiching it in the thickness direction of the first sheet-like member 11 and the third sheet-like member 14. Therefore, the cell sheet 101 is held by being sandwiched between the two mesh-like sheet members 211 and 212, and is fixed in a positioned state inside the three-dimensional containing section 105, and is contained in the three-dimensional containing section 105. Furthermore, by forming the three-dimensional containing section 105 into a three-dimensional shape, the thickness in the Z direction can be formed to be the same at any position in the X direction and the Y direction. As a result, when the cell sheet 101 is positioned in the three-dimensional containing section 105, the distance from the outer surface of the three-dimensional containing section 105 to the cell sheet 101 contained inside the three-dimensional containing section 105 can be made constant. Therefore, the cell sheet 101 contained in the cryopreservation container 1 can be frozen and thawed more uniformly.

[0066] (3) The cell sheet 101 is formed in a sheet shape, and has two mesh-like sheet members 211, 212 that sandwich and hold the cell sheet 101 in the thickness direction of the first sheet-like member 11 and the third sheet-like member 14. This allows the cell sheet 101 to be sandwiched and held between the two mesh-like sheet members 211, 212 in a state in which the cell sheet 101 is in contact with the cryoprotective liquid 102 inside the three-dimensional storage section 105. Therefore, with a simple configuration, the cell sheet 101 can be safely stored while being protected by the cryoprotective liquid 102 inside the three-dimensional storage section 105.

[0067] (4) The two mesh-like sheet members 211, 212 hold the cell sheet 101 inside the three-dimensional containing section 105, and the outer peripheral edge is sandwiched and held between the first sheet-like member 11 and the third sheet-like member 14. Thus, the cell sheet 101 can be held by the two mesh-like sheet members 211, 212 in a stable state with a simple configuration.

[0068] Next, a cryopreservation container 1A of a second embodiment will be described with reference to Figures 10 to 18. The second embodiment differs from the first embodiment mainly in that the third sheet-like member 14A does not have a three-dimensional shaped portion and that the shapes of the two mesh-like sheet members 213, 214 of the cell sheet holding structure 21A are different. In the second embodiment, the same components as those of the first embodiment are denoted by the same reference numerals and description thereof will be omitted.

[0069] 10 to 16, the container body 10A includes a first sheet-like member 11, a heat-resistant sheet-like member 12, a second sheet-like member 13, a third sheet-like member 14A, a weak seal portion 15, a strong seal portion 16, and a sealing seal portion 17. The third sheet-like member 14A does not have a three-dimensional shape portion and is formed into a flat sheet shape as a whole.

[0070] As shown in Figures 13 to 15, the first sheet-like member 11, the heat-resistant sheet-like member 12, the second sheet-like member 13 and the third sheet-like member 14A are stacked and arranged in this order from one side Z1 to the other side Z2 in the Z direction, similar to the first embodiment.

[0071] The first sheet-like member 11 has a bottom-side peripheral sheet-like portion 111 (a three-dimensionally shaped portion peripheral sheet-like portion) and a bottom-side three-dimensionally shaped portion 112 (a three-dimensionally shaped portion) as in the first embodiment. The third sheet-like member 14A is formed in a planar sheet shape without a three-dimensionally shaped portion, unlike the first embodiment. The third sheet-like member 14A is arranged by stacking on the other side Z2 in the Z direction of the first sheet-like member 11 with the opening of the bottom-side three-dimensionally shaped portion 112 of the first sheet-like member 11 facing one side Z1 in the Z direction of the third sheet-like member 14A. With the bottom-side three-dimensionally shaped portion 112 of the first sheet-like member 11 and the third sheet-like member 14A arranged facing each other, the bottom-side three-dimensionally shaped portion 112 of the first sheet-like member 11 and the third sheet-like member 14A are stacked in the Z direction to form a three-dimensional storage portion 106 having a thickness in the Z direction.

[0072] The three-dimensional storage section 106 is formed by the bottom three-dimensional shaped section 112 of the first sheet-like member 11 and the third sheet-like member 14A on the inside in the planar direction of the first sheet-like member 11 and the third sheet-like member 14A, and is three-dimensionally formed with a thickness in the thickness direction of the first sheet-like member 11 and the third sheet-like member 14A so as to protrude to one side Z1 in the Z direction of the container body 10A of the cryopreservation container 1A.

[0073] The three-dimensional containing section 106 contains biological samples such as the cell sheet 101 and other cells in a state held by the cell sheet holding structure 21A. The three-dimensional containing section 106 has an outer shape formed with a thickness in the Z direction, and has a circular outer shape in a plan view. As shown in FIG. 14, the three-dimensional containing section 106 has a rectangular cross-sectional shape extending in the Y direction when cut along the X direction, and as shown in FIG. 15, the three-dimensional containing section 106 has a rectangular cross-sectional shape extending in the Y direction when cut along the Y direction.

[0074] The cell sheet holding structure 21A has two mesh sheet members 213 and 214 (mesh sheet members). Both of the two mesh sheet members 213 and 214 are formed in a mesh-like sheet shape, and are composed of a first mesh sheet member 213 (first mesh sheet member) and a second mesh sheet member 214 (second mesh sheet member).

[0075] The two mesh-like sheet members 213, 214 are arranged such that the first mesh-like sheet member 213 is arranged on one side Z1 in the Z direction and the second mesh-like sheet member 214 is arranged on the other side Z2 in the Z direction, and are arranged stacked in the Z direction with the cell sheet 101 sandwiched and held between the second sheet-like member 13 and the third sheet-like member 14A in the Z direction. By arranging the cell sheet 101 between the first mesh-like sheet member 213 and the second mesh-like sheet member 214, the first mesh-like sheet member 213 and the second mesh-like sheet member 214 hold the cell sheet 101 accommodated in the three-dimensional accommodation section 106 by sandwiching it in the thickness direction of the first sheet-like member 11 and the third sheet-like member 14A. The cell sheet 101 held by the two mesh-like sheet members 213, 214 of the cell sheet holding structure 21A is accommodated in the three-dimensional accommodation section 106. The two mesh sheet members 213 and 214 hold the cell sheet 101 at the center in the Z direction, which is the thickness direction of the three-dimensional containing section .

[0076] As shown in Fig. 13 to Fig. 15, the first mesh-like sheet member 213 and the second mesh-like sheet member 214 each have an upper outer peripheral sheet-like portion 213a, 214a and a stepped circular recess 213b, 214b arranged inside the upper outer peripheral sheet-like portion 213a, 214a. The upper outer peripheral sheet-like portion 213a, 214a is formed in a sheet shape arranged around the bottom three-dimensional shape portion 112 constituting the three-dimensional storage portion 106, and has a rectangular outer shape in a plan view. The upper outer peripheral sheet-like portion 213a, 214a is formed in a planar shape smaller than the outer size of the second sheet-like member 13 and the third sheet-like member 14A.

[0077] The stepped circular recesses 213b, 214b are formed in a circular shape in a plan view at the center of the inner side of the upper outer peripheral sheet-like portions 213a, 214a, and are each formed to be recessed in a stepped shape on the same one side Z1 in the Z direction. The stepped circular recesses 213b, 214b each have a circular bottom plate 213c, 214c formed in a circular sheet shape, and a cylindrical stepped wall portion 213d, 214d. The stepped wall portion 213d, 214d connects the circular bottom plate 213c, 214c to the upper outer peripheral sheet-like portions 213a, 214a.

[0078] The first mesh-like sheet member 213 and the second mesh-like sheet member 214 are stacked in this order from one side Z1 to the other side Z2 in the Z direction, so that they are stacked in the Z direction with their respective stepped circular recesses 213b, 214b and upper outer peripheral sheet-like portions 213a, 214a overlapping each other.

[0079] The circular bottom plate 214c of the stepped circular recess 214b of the second mesh-like sheet member 214 is formed in a circular shape slightly smaller in diameter than the circular bottom plate 213c of the stepped circular recess 213b of the first mesh-like sheet member 213 so that the stepped circular recess 214b of the second mesh-like sheet member 214 can be stacked on the other side Z2 of the stepped circular recess 213b of the first mesh-like sheet member 213, and the stepped wall portion 214d of the stepped circular recess 214b of the second mesh-like sheet member 214 is formed to be slightly smaller in height in the Z direction than the stepped wall portion 213d of the stepped circular recess 213b of the first mesh-like sheet member 213.

[0080] This results in the cell sheet 101 being sandwiched between the circular bottom plate 213c of the stepped circular recess 213b of the first mesh-like sheet member 213 and the circular bottom plate 214c of the stepped circular recess 214b of the second mesh-like sheet member 214, with the stepped circular recess 213b of the first mesh-like sheet member 213 and the stepped circular recess 214b of the second mesh-like sheet member 214 being stacked and arranged.

[0081] The two mesh-like sheet members 213, 214 hold the cell sheet 101 by sandwiching it between the circular bottom plates 213c, 214c of the stepped circular recesses 213b, 214b inside the three-dimensional containing section 106. The upper outer peripheral sheet-like members 213a, 214a provided on the outer periphery of the stepped circular recesses 213b, 214b are sandwiched and held between the second sheet-like member 13 and the third sheet-like member 14A in an overlapping state.

[0082] The height in the Z direction of the step wall 214d of the stepped circular recess 214b of the second mesh-like sheet member 214 and the step wall 213d of the stepped circular recess 213b of the first mesh-like sheet member 213 are set to a height at which they are disposed in the center in the Z direction of the three-dimensional containing section 106 formed by the first sheet-like member 11 and the third sheet-like member 14A. Therefore, the circular bottom plates 213c, 214c of the stepped circular recesses 213b, 214b of the two mesh-like sheet members 213, 214 are disposed in the center in the Z direction inside the three-dimensional containing section 106 in a state where they are stacked in the Z direction. As a result, inside the three-dimensional containing section 106, the cell sheet 101 sandwiched between the circular bottom plates 213c, 214c of the stepped circular recesses 213b, 214b is disposed in the center in the Z direction inside the three-dimensional containing section 106. Therefore, the two mesh sheet members 213 and 214 hold the cell sheet 101 at the center in the Z direction, which is the thickness direction of the three-dimensional containing section .

[0083] A liquid introduction tube 20 disposed at one end of the container body 10 in the Y direction communicates with the three-dimensional containing section 106 on one side in the Y direction. With the cell sheet 101 contained in the three-dimensional containing section 106, a cryoprotectant 102 (see FIG. 14 ) is introduced into the three-dimensional containing section 106 by the liquid introduction tube 20.

[0084] When storing a biological sample such as a cell sheet 101 in the cryopreservation container 1A, the cryopreservation container 1A is produced in a factory by stacking a first sheet-like member 11, a heat-resistant sheet-like member 12, and a second sheet-like member 13, and sealing them at the weak seal portion 15 and the strong seal portion 16, as shown in Figs. 14 to 16, in the same manner as in the first embodiment. When the cryopreservation container 1A is shipped from the factory, the third sheet-like member 14A is shipped in a state in which only the liquid introduction tube side sealing portion 18 is sealed. Furthermore, when shipped from the factory, the two mesh-like sheet members 213, 214 of the cell sheet holding structure 21A and the cell sheet 101 are not placed between the second sheet-like member 13 and the third sheet-like member 14A.

[0085] After shipment from the factory, as in the first embodiment, the user places two mesh sheet members 213, 214 sandwiching a cell sheet 101 between the second sheet-like member 13 and the third sheet-like member 14A, as shown in Figures 14 and 15, and seals the second sheet-like member 13 and the third sheet-like member 14A with the sealing portion 17 at the portions where they are laminated to the heat-resistant sheet-like member 12.

[0086] In this case, since the heat-resistant sheet-like member 12 is disposed on one side Z1 of the second sheet-like member 13 and the third sheet-like member 14A, the first sheet-like member 11 does not melt due to heat, and the second sheet-like member 13 and the third sheet-like member 14A are sealed together by the sealing portion 17. When the sealing portion 17 seals the weak seal portion 15 and the strong seal portion 16, the peripheral portions of the two mesh-like sheet members 213, 214 of the cell sheet holding structure 21A disposed in the heat-resistant sheet-like member 12 from the inside to the middle of the width direction are simultaneously sealed.

[0087] 16, a cryoprotectant liquid 102 is introduced into the three-dimensional container 106, and the opening of the liquid introduction tube 20 is sealed. In this manner, the user can contain a biological sample, such as a cell sheet 101 or other cells, in the cryopreservation container 1A and seal it. The cryopreservation container 1A containing the cell sheet, other cells, etc. is then frozen and preserved.

[0088] Here, the cell sheet 101 held by the two mesh-like sheet members 213, 214 is disposed inside the three-dimensional containing section 106 at the center in the Z direction of the three-dimensional containing section 106. As a result, the two mesh-like sheet members 213, 214 hold the cell sheet 101 at the center in the Z direction, which is the thickness direction of the three-dimensional containing section 106. Therefore, the distance from the cell sheet 101 disposed inside the three-dimensional containing section 106 to the outer surface of one side Z1 and the outer surface of the other side Z2 in the Z direction of the three-dimensional containing section 106 can be set to the same distance, so that the cell sheet 101 contained in the cryopreservation container 1 can be frozen uniformly.

[0089] In the second embodiment, when the cryopreservation container 1A is opened and removed, the frozen cryopreservation container 1A is first thawed as in the first embodiment. Here, the two mesh-like sheet members 213, 214 hold the cell sheet 101 at the center in the Z direction, which is the thickness direction of the three-dimensional containing section 106. Therefore, the distance from the cell sheet 101 placed inside the three-dimensional containing section 106 to the outer surface of one side Z1 and the outer surface of the other side Z2 in the Z direction of the three-dimensional containing section 106 can be set to the same distance, so that the cell sheet 101 contained in the cryopreservation container 1A can be thawed uniformly.

[0090] 17 and 18, the outer peripheral edge 19 (end portion 11b of the first sheet-like member 11, end portion 13b of the second sheet-like member 13, end portion 14b of the third sheet-like member 14A) including the portion where the strong seal portion 16 of the container body 10A is formed is cut off with a cutting tool such as scissors. After that, the heat-resistant sheet member 12 disposed between the first sheet-like member 11 and the second sheet-like member 13 is removed.

[0091] 17, after the outer peripheral edge portion 19 is cut off, the first sheet-like member 11 and the second sheet-like member 13 are pulled to one side Z1 and the other side Z2 in the Z direction so that they are separated from each other, and the first sheet-like member 11 and the second sheet-like member 13 are peeled off at the weak seal portion 15. The weak seal portion 15 has a seal strength that prevents the cryoprotectant liquid 102 injected into the three-dimensional storage portion 106 from leaking out, has a seal strength weaker than that of the sealing seal portion 17, and is a seal portion sealed by heat with a seal strength that can be opened by hand, and the second sheet-like member 13 is strongly sealed to the third sheet-like member 14A side, so that the first sheet-like member 11 and the second sheet-like member 13 can be easily peeled off at the weak seal portion 15. After opening at the weak seal portion 15, the two mesh sheet members 213 and 214 are partially torn to remove the cell sheet 101, in the same manner as in the first embodiment.

[0092] This allows the container body 10A to be opened and the cell sheet 101 contained in the three-dimensional containing portion 106 to be removed. Before cutting the outer periphery 19 including the strong seal portion 16, the outer periphery is sealed by the strong seal portion 16, and after removing the strong seal portion 16, the first sheet-like member 11 and the second sheet-like member 13 can be peeled away from each other to open the container at the weak seal portion 15. This prevents the cell sheet 101 contained in the three-dimensional containing portion 106 from being contaminated.

[0093] In the second embodiment, in addition to the same advantages as those (1) to (4) of the first embodiment, the following advantages can be achieved.

[0094] (5) The two mesh-like sheet members 213, 214 are composed of a first mesh-like sheet member 213 and a second mesh-like sheet member 214, and the stepped circular recess 213b of the first mesh-like sheet member 213 and the stepped circular recess 214b of the second mesh-like sheet member 214 are stacked in a state in which the cell sheet 101 is sandwiched between the circular bottom plate 213c of the stepped circular recess 213b of the first mesh-like sheet member 213 and the circular bottom plate 214c of the stepped circular recess 214b of the second mesh-like sheet member 214. This allows the distances from the cell sheet 101 placed inside the three-dimensional containing section 106 to the outer surface of one side Z1 and the outer surface of the other side Z2 in the Z direction of the three-dimensional containing section 106 to be the same. Therefore, the cell sheet 101 contained in the cryopreservation container 1A can be frozen and thawed more uniformly.

[0095] Although the preferred embodiments of the cryopreservation container of the present invention have been described above, the present invention is not limited to the above-described embodiments and can be modified as appropriate.

[0096] For example, in the above embodiment, the three-dimensional containing sections 105 and 106 are formed to have a circular shape in a plan view, but this is not limited thereto. For example, the three-dimensional containing sections 105 and 106 may be formed to have an elliptical or polygonal shape in a plan view.

[0097] In the above embodiment, the shape of the cell sheet 101 in a planar view is circular, but this is not limiting. For example, the shape of the cell sheet 101 in a planar view may be elliptical or polygonal.

[0098] In the above embodiment, the biological sample is constituted by a sheet-shaped cell sheet, but is not limited thereto. For example, the biological sample does not have to be formed in a sheet shape as long as it can be held at the center in the thickness direction of the three-dimensional container section.

[0099] In the above embodiment, the biological sample is held between two mesh-like sheet members at the center of the thickness direction of the three-dimensional container, but this is not limited thereto. The means for holding the biological sample at the center of the thickness direction of the three-dimensional container does not have to be a method of holding the biological sample between two mesh-like sheet members, and may be, for example, a method of fixing the biological sample on a single sheet, or a method of providing a pedestal (e.g., a tube, etc.) on a bottom sheet member arranged on the bottom side and holding the biological sample on the pedestal. [Explanation of symbols]

[0100] 1,1A cryopreservation container 11 First sheet-like member (bottom sheet-like member) 14, 14A Third sheet-like member (lid-side sheet-like member) 21, 21A Cell sheet holding structure (biological sample holding part) 101 Cell Sheet (Biological Sample) 105,106 Three-dimensional storage section 111 Bottom side peripheral sheet-like portion (first three-dimensional shape portion peripheral sheet-like portion) 112 Bottom three-dimensional shape part (first three-dimensional shape part) 141 Lid side surrounding sheet-like portion (second three-dimensional shape portion surrounding sheet-like portion) 142 Lid side three-dimensional shape part (second three-dimensional shape part) 211, 212 Mesh-like sheet member (mesh-like sheet portion) 213 First mesh-like sheet member (mesh-like sheet member, mesh-like sheet portion) 214 Second mesh-like sheet member (mesh-like sheet member, mesh-like sheet portion)

Claims

1. A bottom sheet-like member; A lid-side sheet-like member disposed by being laminated on the bottom-side sheet-like member; a three-dimensional storage section formed by the bottom sheet-like member and the lid sheet-like member on the inner side in the planar direction of the bottom sheet-like member and the lid sheet-like member, the three-dimensional storage section having a thickness in the thickness direction of the bottom sheet-like member and the lid sheet-like member, and a biological sample is stored therein; A cryopreservation container comprising: a biological sample holding section that holds the biological sample contained in the three-dimensional container section at the center in the thickness direction of the three-dimensional container section.

2. The cryopreservation container according to claim 1 , wherein the biological sample holder holds the biological sample by sandwiching it in a thickness direction of the bottom sheet member and the lid sheet member.

3. The biological sample is formed into a sheet shape, The cryopreservation container according to claim 1 or 2, wherein the biological sample holding section has two mesh sheet sections that hold the sheet-like biological sample by sandwiching it in the thickness direction between the bottom sheet member and the lid sheet member.

4. The freezing storage container of claim 3, wherein the two mesh sheet portions hold the biological sample inside the three-dimensional storage section arranged on the inner side of the planar direction of the bottom sheet member and the lid sheet member, and the planar outer peripheral edges of the bottom sheet member and the lid sheet member are held by being sandwiched between the bottom sheet member and the lid sheet member.

5. the bottom sheet-like member has a first three-dimensional shape portion formed in a concave shape with an open side toward the lid sheet-like member, and a first three-dimensional shape portion surrounding sheet-like member disposed around the first three-dimensional shape portion, the cover-side sheet-like member has a second three-dimensional shape portion formed in a concave shape with an open bottom-side sheet-like member side, and a second three-dimensional shape portion surrounding sheet-like member disposed around the second three-dimensional shape portion, The cryopreservation container according to claim 1 or 2, wherein the three-dimensional storage section is configured by arranging open portions of the first three-dimensional shape section and the second three-dimensional shape section facing each other.

6. The two mesh sheet portions are composed of a first mesh sheet portion having a first concave portion that is concave in a stepped shape on one side in the thickness direction of the bottom sheet member and the lid sheet member, and a second mesh sheet portion having a second concave portion that is concave in a stepped shape on one side in the same thickness direction as the first concave portion is concave in the thickness direction of the bottom sheet member and the lid sheet member, and the first concave portion and the second concave portion are stacked and arranged with the biological sample sandwiched between the bottom of the first concave portion and the bottom of the second concave portion.

Images