Culture vessel and method for manufacturing the culture vessel

Abstract

To provide a culture vessel that allows the size of a culture space to be changed.SOLUTION: A culture vessel comprises a main body having a culture area and an expansion area on a culture surface, and a partition wall that covers the expansion area from above. The removal of the partition wall from the expansion area opens the expansion area.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a culture vessel and a method for manufacturing the culture vessel.

Background Art

[0002] Conventionally, as disclosed in Patent Document 1, cell culture using a culture vessel has been performed. Such a culture vessel has a culture space for culturing cells. And cell culture is performed with cells and a culture solution placed in the culture space.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] The culture space of the culture vessel as described above has a predetermined size. Therefore, the size of the culture space cannot be changed according to the progress of cell culture.

[0005] An object of the present invention is to provide a culture vessel and a method for manufacturing the culture vessel capable of changing the size of the culture space.

Means for Solving the Problems

[0006] One aspect of the culture vessel according to the present invention is a main body having a culture region and an expansion region on a culture surface, and a partition wall covering the expansion region from above, wherein when the partition wall is removed from the expansion region, the expansion region is opened.

[0007] One aspect of the method for manufacturing a culture vessel according to the present invention is a step of providing a main body having a culture region and an expansion region on a culture surface, providing a partition portion at a position facing the expansion region; adhering the partition portion to the expansion region, and including.

Advantages of the Invention

[0008] According to the present invention, a culture vessel capable of changing the size of a culture region and a method for manufacturing the culture vessel can be provided.

Brief Description of the Drawings

[0009]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Modes for Carrying Out the Invention

[0010] Hereinafter, the culture vessel according to the present invention will be described in detail with reference to the drawings. Note that the culture vessel and the method for manufacturing the culture vessel described below are examples of the culture vessel and the method for manufacturing the culture vessel according to the present invention, and the present invention is not limited by the embodiments described later.

[0011] [Embodiment 1] With reference to FIGS. 1 to 4, a culture vessel and a method for manufacturing the culture vessel according to Embodiment 1 of the present invention will be described.

[0012] The culture vessel 1 is used, for example, for culturing cells derived from humans. Such a culture vessel 1 has a culture space for accommodating cells. The culture vessel 1 is accommodated in the culture space of a culture device (for example, an incubator) in a state where cells to be cultured (hereinafter referred to as "target cells") are accommodated in the culture space.

[0013] The culture vessel 1 according to the present embodiment can be used, for example, for culturing spheroids (cell aggregates) of target cells. Note that the culture vessel 1 does not necessarily need to be used in a state of being accommodated in a culture device. The culture vessel 1 may be used in various situations according to the target cells. Further, the shape of the culture vessel is not limited to the shape of the culture vessel 1.

[0014] Hereinafter, the specific configuration of the culture vessel 1 will be described. In the following description of the culture vessel 1, for convenience of explanation, the first direction, the second direction, and the third direction shown in each figure are used. The culture vessel 1 is box-shaped. The first direction of the culture vessel 1 coincides with the lateral direction of the culture vessel 1 (in other words, the left-right direction). The second direction of the culture vessel 1 coincides with the longitudinal direction of the culture vessel 1 (in other words, the front-rear direction). The third direction of the culture vessel 1 coincides with the vertical direction of the culture vessel 1.

[0015] Note that the shape of the culture vessel is not limited to the shape of the culture vessel 1. For example, the culture vessel may have a circular shape in plan view. Plan view means looking at the culture vessel from above. The shape of the culture vessel in plan view is the outer shape of the culture vessel when viewed from above.

[0016] As shown in FIG. 1, the culture vessel 1 has a main body portion 2 and a partition portion 3.

[0017] In the culture vessel 1 according to the present embodiment, the size of the culture space changes between the state where the partition portion 3 is supported by the main body portion 2 and the state where the partition portion 3 is removed from the main body portion 2. Specifically, when the partition portion 3 is removed from the main body portion 2, the culture space expands. Note that the culture space may be regarded as the space surrounded by the bottom surface portion 21 and the peripheral wall portion 22 of the main body portion 2.

[0018] In the following description, the state where the partition portion 3 is supported by the main body portion 2 is referred to as the reference state of the culture vessel 1. Also, the state where the partition portion 3 is removed from the main body portion 2 is referred to as the expanded state of the culture vessel 1. Hereinafter, the specific configurations of the main body portion 2 and the partition portion 3 will be described.

[0019] FIG. 1 is a perspective view of the culture vessel 1 in the reference state. Also, FIG. 2 is a plan view of the culture vessel 1 in the reference state. Also, FIG. 3 is a plan view of the culture vessel 1 in the expanded state.

[0020] As shown in FIG. 1, the main body portion 2 has a bottom surface portion 21 and a peripheral wall portion 22. The main body portion 2 has a box shape with an open upper end. Note that the main body portion may have a lid portion (not shown) that closes the upper end.

[0021] The bottom surface portion 21 is plate-shaped. In the case of the present embodiment, the shape of the bottom surface portion 21 in plan view is a quadrangle. However, the shape of the bottom surface portion in plan view is not limited to a quadrangle. For example, the shape of the bottom surface portion in plan view may be a circle, an ellipse, or a polygon other than a quadrangle.

[0022] The bottom surface portion 21 is made of, for example, a synthetic resin. Examples of the synthetic resin constituting the bottom surface portion 21 include polystyrene and polyolefin. The polyolefin may be a cyclic olefin-based (co)polymer, a 4-methyl-1-pentene-based (co)polymer, polypropylene, or polyethylene. From the viewpoint of gas permeability, a 4-methyl-1-pentene-based (co)polymer is preferable. Also, from the viewpoint of heat resistance, polystyrene, polypropylene, or a cyclic olefin-based (co)polymer is preferable.

[0023] The 4-methyl-1-pentene-based 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.

[0024] The other α-olefin may be linear, may have a branch, or may be a cyclic olefin.

[0025] 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-olefins 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. Among these, ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, and 1-decene are preferable, and ethylene and propylene are more preferable.

[0026] From the viewpoint of enhancing mechanical strength, the intrinsic viscosity [η] of the 4-methyl-1-pentene polymer in decalin at 135°C is preferably 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, still 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.

[0027] The 4-methyl-1-pentene polymer has a density measured in accordance with ASTM D1505 (water substitution method) of 3 0.810 g / cm 3 or more and 3 0.850 g / cm 3 or less, more preferably 3 0.820 g / cm 3 or more and 0.850 g / cm or less, and still more preferably 0.830 g / cm or more and 0.850 g / cm or less.

[0028] 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 (International Publication Nos. 01 / 53369, 01 / 027124, Japanese Patent Laid-Open Nos. 3-193796, and 02-41303, etc.), and an olefin polymerization catalyst containing a metallocene compound (International Publication No. 2011 / 055803).

[0029] The oxygen permeability at 23°C of the bottom surface portion 21 is preferably 3 7500 cm 2 / (m 3 ·24 h·atm) or more and 150000 cm 2 / (m 3 ·24 h·atm) or less. Further, the oxygen permeability at 23°C of the bottom surface portion 21 is more preferably 2 8000 cm 3 / (m 2 ·24 h·atm) or more and 60000 cm 3 / (m2 ·24 h·atm) or more, 40,000 cm 3 / (m 2 ·24 h·atm) or less is more preferable.

[0030] The higher the oxygen permeability, the easier it is to promote cell proliferation and growth. Also, when the oxygen permeability is moderately low, the cell adhesion is good, so the culture container 1 can be applied when culturing a wider variety of cells.

[0031] As shown in FIG. 3, the bottom surface portion 21 has a culture region 211 and an expansion region 212 on the upper surface. The upper surface of the bottom surface portion 21 corresponds to an example of a culture surface. The culture region 211 and the expansion region 212 are planar regions on the upper surface of the bottom surface portion 21. The upper surface of the bottom surface portion 21 corresponds to an example of the culture surface of the main body portion.

[0032] The culture region 211 is a region that is open upward on the upper surface of the bottom surface portion 21. Specifically, the culture region 211 is a region that is open upward in the reference state and the expanded state of the culture container 1. In the culture region 211, target cells, a culture solution, and / or a drug, etc. are arranged during cell culture.

[0033] In addition, the culture region 211 may be regarded as a region where substances related to cell culture are arranged during cell culture. That is, in the culture region 211, target cells may not be arranged during cell culture, and only a culture solution or a drug may be arranged.

[0034] In the reference state of the culture container 1, a first culture space is formed above the culture region 211. The first culture space is a space surrounded by the bottom surface portion 21 and the partition portion 3 above the culture region 211. In the reference state of the culture container 1, the first culture space corresponds to the culture space of the culture container 1.

[0035] In the case of this embodiment, the culture region 211 is divided into a plurality of divided culture regions Ra and Rb by an expansion region 212 (in other words, a partition portion 3) described later. In other words, the region obtained by combining all of the divided culture regions Ra and Rb is the culture region 211. The divided culture regions Ra and Rb respectively correspond to examples of a first section and a second section.

[0036] The divided culture region Ra and the divided culture region Rb are not in communication in the reference state of the culture vessel 1. Therefore, the target cells and the culture solution accommodated in one of the divided culture regions (for example, the divided culture region Ra) of the divided culture region Ra and the divided culture region Rb cannot move to the other divided culture region (for example, the divided culture region Rb) of the divided culture region Ra and the divided culture region Rb in the reference state of the culture vessel 1.

[0037] The expansion region 212 is a region covered from above by a partition portion 3 (described later) on the upper surface of the bottom portion 21. In the case of this embodiment, the expansion region 212 is a region between the divided culture regions Ra and Rb on the upper surface of the bottom portion 21.

[0038] Such an expansion region 212 is opened when the partition portion 3 is removed from the expansion region 212. That is, the expansion region 212 is a region that is opened in the expanded state of the culture vessel 1. When the expansion region 212 is opened, the adjacent divided culture regions Ra and Rb separated by the partition portion 3 communicate with each other.

[0039] Incidentally, in the expanded state of the culture vessel 1, the space above the expansion region 212 is referred to as a second culture space. In the expanded state of the culture vessel 1, the space obtained by combining the first culture space above the culture region 211 and the second culture space above the expansion region 212 corresponds to the culture space of the culture vessel 1. Therefore, the culture space in the expanded state of the culture vessel 1 is wider than the culture space in the reference state of the culture vessel 1 by the space corresponding to the second culture space.

[0040] The peripheral wall portion 22 is in the shape of a rectangular frame. The peripheral wall portion 22 is provided so as to surround the bottom surface portion 21. Specifically, the peripheral wall portion 22 is provided along the outer peripheral edge of the bottom surface portion 21. The peripheral wall portion 22 extends upward from the outer peripheral edge of the bottom surface portion 21.

[0041] In the case of this embodiment, the peripheral wall portion 22 and the bottom surface portion 21 are integrally formed in a non-separable state. The material constituting the peripheral wall portion 22 is the same as the material (for example, synthetic resin) constituting the bottom surface portion 21. Note that the peripheral wall portion 22 may be omitted.

[0042] The partition portion 3 is plate-shaped and is provided in the extension region 212 of the bottom surface portion 21. Specifically, the lower surface of the partition portion 3 is adsorbed to the extension region 212. The adsorption force between the extension region 212 and the partition portion 3 is such that the culture solution does not flow between at least the divided culture regions Ra and Rb.

[0043] In the case of this embodiment, an adhesive layer for adhering the partition portion 3 and the extension region 212 is not provided between the lower surface of the partition portion 3 and the extension region 212. Therefore, during cell culture, the adhesive does not dissolve in the culture solution.

[0044] Further, both end faces of the partition portion 3 (specifically, both end faces in the first direction) are adsorbed to the inner surface of the peripheral wall portion 22. An adhesive layer for adhering both end faces of the partition portion 3 and the inner surface of the peripheral wall portion 22 is not provided between both end faces of the partition portion 3 and the inner surface of the peripheral wall portion 22.

[0045] The adsorption force between both end faces of the partition portion 3 (specifically, both end faces in the first direction) and the inner surface of the peripheral wall portion 22 is such that the culture solution does not flow between at least the divided culture regions Ra and Rb.

[0046] The partition portion 3 is made of synthetic resin. Examples of the synthetic resin constituting the partition portion 3 include polystyrene and polyolefin. The polyolefin may be a cyclic olefin-based (co)polymer, a 4-methyl-1-pentene-based (co)polymer, polypropylene, or polyethylene. From the viewpoint of gas permeability, a 4-methyl-1-pentene-based (co)polymer is preferable. Further, from the viewpoint of heat resistance, polystyrene, polypropylene, or a cyclic olefin-based (co)polymer is preferable.

[0047] The 4-methyl-1-pentene-based 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.

[0048] The other α-olefin may be linear, branched, or a cyclic olefin.

[0049] 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. Among these, ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, and 1-decene are preferable, and ethylene and propylene are more preferable.

[0050] From the viewpoint of enhancing mechanical strength, the intrinsic viscosity [η] of the 4-methyl-1-pentene polymer in decalin at 135 °C is preferably 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, still 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.

[0051] The density of the 4-methyl-1-pentene polymer measured in accordance with ASTM D1505 (water substitution method) is 3 0.810 g / cm 3 or more and 3 0.850 g / cm 3 or less, more preferably 3 0.820 g / cm 3 or more and

[0052] 0.850 g / cm

[0053] The oxygen permeability at 23 °C of the partition portion 3 is preferably 3 7500 cm 2 / (m 3 ·24 h·atm) or more and 2 150000 cm 3 / (m 2 ·24 h·atm) or less. Also, the oxygen permeability at 23 °C of the partition portion 3 is more preferably 3 8000 cm 2 / (m 3 ·24 h·atm) or more and2 ·24 h·atm) or more, 40,000 cm 3 / (m 2 ·24 h·atm) or less is more preferable.

[0054] The higher the oxygen permeability, the easier it is to promote cell proliferation and growth. Also, when the oxygen permeability is moderately low, the cell adhesiveness becomes good, so the culture vessel 1 can be applied when culturing a wider variety of cells.

[0055] Next, an example of the usage method of the culture vessel 1 will be described. The culture vessel 1 is used, for example, in a test for evaluating cell-cell interactions of different cells. The following description is also an example of a cell culture method carried out using the culture vessel 1.

[0056] First, the operator prepares the culture vessel 1 in the reference state as shown in FIGS. 1 and 2. Then, the operator accommodates the first target cells in the divided culture region Ra of the culture vessel 1. Also, the operator accommodates the second target cells in the divided culture region Rb of the culture vessel 1. The first target cells and the second target cells may be different types of cells or the same type of cells.

[0057] The operator accommodates a culture solution, a drug, etc. suitable for culturing the first target cells in the divided culture region Ra. Also, the operator accommodates a culture solution, a drug, etc. suitable for culturing the second target cells in the divided culture region Rb. Then, the operator accommodates the culture vessel 1 in the culture space of a culture device (for example, an incubator), for example. Such a process corresponds to an example of the first step in the cell culture method. The first step in the cell culture method is a step of starting cell culture in the culture region 211 of the culture surface.

[0058] Next, after a predetermined time has elapsed since the start of cell culture, the operator takes out the culture vessel 1 from the culture device (for example, an incubator).

[0059] Before performing the partition removal operation described below, the operator may discard the culture solutions contained in the divided culture regions Ra and Rb. Whether or not to perform the operation of discarding the culture solution may be appropriately determined according to the test conditions.

[0060] In this state, in the divided culture region Ra, the first target cells are growing. Also, in the divided culture region Rb, the second target cells are growing.

[0061] Next, the operator removes the partition portion 3 from the culture vessel 1. In other words, the operator removes the partition portion 3 from the extended region 212 of the bottom surface portion 21 in the culture vessel 1. This operation is referred to as the partition removal operation. When performing the partition removal operation, the operator holds down the main body portion 2 and pulls up the partition portion 3 upward. Such a process corresponds to an example of the second step in the cell culture method. The second step in the cell culture method is a step of removing the partition portion 3 and opening the extended region 212.

[0062] Then, the extended region 212 (see FIG. 3) is opened, and the divided culture region Ra and the divided culture region Rb communicate with each other. And the culture vessel 1 becomes the extended state shown in FIG. 3. In the extended state of the culture vessel 1, the extended region 212 is opened, and a second culture space is formed above the extended region 212. In this way, the culture space of the culture vessel 1 is expanded.

[0063] In the extended state of the culture vessel 1, the operator may further continue culturing the cells in the culture region 211 and the extended region 212. Such a process corresponds to an example of the third step in the cell culture method.

[0064] In addition, the operator may administer a drug to the cells contained in the culture vessel 1 at an appropriate timing. The step of administering such a drug may be performed before the first step in the cell culture method. Also, the step of administering such a drug may be performed between the first step and the second step in the cell culture method.

[0065] Furthermore, the step of administering the agent may be carried out between the second step and the third step in the cell culture method. Alternatively, the step of administering the agent may be carried out after the third step in the cell culture method.

[0066] Furthermore, in the expanded state of the culture vessel 1, the operator may observe the movement of the first target cells grown in the divided culture region Ra and the second target cells grown in the divided culture region Rb.

[0067] For example, the first target cells and the second target cells move toward the expansion region 212 based on the reaction to the agent and / or the cell-cell interaction between the first target cells and the second target cells. Such a property of the cells is referred to as cell migration. The operator can evaluate the cell migration by confirming the movement status of the first target cells and the second target cells.

[0068] In addition, the agent may be administered to the first target cells and the second target cells by accommodating it in each of the divided culture regions Ra and Rb before the start of cell culture as described above, or before or after removing the partition portion 3, or simultaneously with removing the partition portion 3. Further, for the purpose of confirming the movement status of the first target cells and the second target cells due to cell migration, a scale for measuring dimensions may be provided in the expansion region 212.

[0069] Note that the above usage example is an example of the method of using the culture vessel 1 according to the present embodiment. Therefore, the method of using the culture vessel 1 is not limited to the above usage example.

[0070] For example, the cells accommodated in the divided culture region Ra of the culture vessel 1 and the cells accommodated in the divided culture region Rb of the culture vessel 1 may be of the same type.

[0071] Alternatively, target cells may be contained only in one of the divided culture regions Ra and Rb. In this case, the other of the divided culture regions Ra and Rb may contain a culture solution and / or a drug used for cell culture.

[0072] In such a usage method, an operator can supply the culture solution and / or the drug contained in the other divided culture region to one divided culture region by removing the partition portion 3 from the culture container 1 at an appropriate timing.

[0073] Next, with reference to FIG. 4, an example of a method for manufacturing the culture container 1 will be described. FIG. 4 is a flowchart regarding the method for manufacturing the culture container 1.

[0074] First, in step S1 of FIG. 4, an operator provides the main body portion 2 at a predetermined position of a manufacturing apparatus (not shown). The main body portion 2 has the bottom surface portion 21 as described above. The bottom surface portion 21 has a culture region 211 and an expansion region 212 on the upper surface. Such a main body portion 2 is manufactured, for example, by injection molding.

[0075] Next, in step S2 of FIG. 4, an operator provides the partition portion 3 at a predetermined position of a manufacturing apparatus (not shown). The predetermined position where the partition portion 3 is provided may be a position facing the expansion region 212.

[0076] Next, in step S3 of FIG. 4, an operator operates the manufacturing apparatus (not shown) to bring the partition portion 3 into close contact (specifically, adsorption) with the expansion region 212. In this way, the culture container 1 is manufactured.

[0077] (Functions and Effects of this Embodiment) According to the culture container 1 having the above-described configuration, the size of the culture region can be changed according to the progress of cell culture.

[0078] Specifically, the culture vessel 1 includes a partition wall portion 3 adsorbed to the expansion region 212 of the bottom surface portion 21. When the partition wall portion 3 is removed from the expansion region 212, the expansion region 212 is opened, and the culture space of the culture vessel 1 is expanded. That is, the size of the culture space changes. Other functions and effects obtained from the culture vessel 1 are as described above.

[0079] [Embodiment 2] Next, with reference to FIGS. 5 to 7, a culture vessel and a method for manufacturing the culture vessel according to Embodiment 2 of the present invention will be described.

[0080] As shown in FIG. 5, the culture vessel 1B has a main body portion 2B and a partition wall portion 3B.

[0081] In the culture vessel 1B according to the present embodiment, the size of the culture space also changes between the state where the partition wall portion 3B is supported by the main body portion 2B and the state where the partition wall portion 3B is removed from the main body portion 2B. Specifically, when the partition wall portion 3B is removed from the main body portion 2B, the culture space expands.

[0082] As shown in FIG. 7, the main body portion 2B has a bottom surface portion 21B. In the case of the present embodiment, the main body portion 2B does not have a configuration corresponding to the peripheral wall portion 22 of the main body portion 2 in the above-described Embodiment 1. In the case of the present embodiment, the main body portion 2B is composed only of the bottom surface portion 21B. Therefore, the main body portion 2B is plate-shaped.

[0083] As shown in FIG. 7, the bottom surface portion 21B is plate-shaped. In the case of the present embodiment, the shape of the bottom surface portion 21B in plan view is a quadrangle. However, the shape of the bottom surface portion in plan view is not limited to a quadrangle. For example, the shape of the bottom surface portion in plan view may be a circle, an ellipse, or a polygon other than a quadrangle.

[0084] The bottom surface portion 21B is made of, for example, a synthetic resin. The synthetic resin constituting the bottom surface portion 21B is the same as the synthetic resin constituting the bottom surface portion 21 in the above-described Embodiment 1.

[0085] As shown in FIG. 7, the bottom surface portion 21B has a culture region 211B and an expansion region 212B on the upper surface. The culture region 211B and the expansion region 212B are planar regions on the upper surface of the bottom surface portion 21B.

[0086] The culture region 211B is a region with an open upper side on the upper surface of the bottom surface portion 21. Specifically, the culture region 211B is a region with an open upper side in the reference state and the expanded state of the culture vessel 1B. In the culture region 211B, target cells, a culture solution, and / or a drug, etc. are arranged during cell culture.

[0087] In the reference state of the culture vessel 1B, a first culture space is formed above the culture region 211B. The first culture space is a space surrounded by the bottom surface portion 21B and the partition portion 3B above the culture region 211B. In the reference state of the culture vessel 1, the first culture space corresponds to the culture space of the culture vessel 1B.

[0088] In the case of this embodiment, the culture region 211B is divided into a plurality of divided culture regions R1 to R15 by the expansion region 212B (in other words, the partition portion 3B) described later. In other words, the region obtained by combining all of the divided culture regions R1 to R15 is the culture region 211B.

[0089] Each of the divided culture regions R1 to R15 corresponds to an example of a first section and a second section. Note that the number of divided culture regions is not limited to the number (that is, 15) of the divided culture regions R1 to R15 in this embodiment.

[0090] Adjacent divided culture regions among the divided culture regions R1 to R15 are not in communication in the reference state of the culture vessel 1B.

[0091] The expansion region 212B is a region covered from above by the partition portion 3B described later on the upper surface of the bottom surface portion 21B. In the case of this embodiment, the expansion region 212B is a lattice-shaped region (the region with diagonal lattice lines in FIG. 7) on the upper surface of the bottom surface portion 21B. In other words, the expansion region 212B is the region between the divided culture regions R1 to R15.

[0092] When such an extended region 212B has the partition wall portion 3B removed therefrom, it is opened. That is, the extended region 212B is an area that is opened in the extended state of the culture vessel 1B. When the extended region 212B is opened, the divided culture regions R1 to R15 communicate with each other.

[0093] In addition, in the extended state of the culture vessel 1B, the space above the extended region 212B is referred to as the second culture space. In the extended state of the culture vessel 1B, the combined space of the first culture space above the culture region 211B and the second culture space above the extended region 212B corresponds to the culture space of the culture vessel 1B. Therefore, the culture space in the extended state of the culture vessel 1B is wider than the culture space in the reference state of the culture vessel 1B by the space corresponding to the second culture space.

[0094] The partition wall portion 3B is lattice-shaped and is provided in the extended region 212B of the bottom surface portion 21B. Specifically, the lower surface of the partition wall portion 3B is adsorbed to the extended region 212B. The adsorption force between the extended region 212B and the partition wall portion 3B is such that the culture solution does not flow at least between the divided culture regions R1 to R15.

[0095] The partition wall portion 3B is made of a synthetic resin. The synthetic resin constituting the partition wall portion 3B is the same as the synthetic resin constituting the partition wall portion 3 in the above-described Embodiment 1.

[0096] Specifically, the partition wall portion 3B has a plurality of horizontal wall portions 31B and a plurality of vertical wall portions 32B.

[0097] Each of the plurality of horizontal wall portions 31B is plate-shaped and parallel to the first direction and the third direction. Each of the plurality of horizontal wall portions 31B has the same length as the length of the bottom surface portion 21B in the first direction. The plurality of horizontal wall portions 31B are arranged at equal intervals in the second direction.

[0098] A pair of horizontal wall portions 31B arranged at both ends in the second direction among the plurality of horizontal wall portions 31B are arranged along the both end edges of the bottom surface portion 21B in the second direction.

[0099] The plurality of vertical wall portions 32B are each plate-shaped and parallel to the second direction and the third direction. Each of the plurality of vertical wall portions 32B has the same length as the length of the bottom surface portion 21B in the second direction. The plurality of vertical wall portions 32B are arranged at equal intervals in the first direction.

[0100] A pair of vertical wall portions 32B arranged at both ends of the plurality of vertical wall portions 32B in the first direction are arranged along both end edges of the bottom surface portion 21B in the first direction.

[0101] The plurality of horizontal wall portions 31B and the plurality of vertical wall portions 32B are arranged perpendicular to each other. The plurality of horizontal wall portions 31B and the plurality of vertical wall portions 32B having the above-described configuration are integrally formed in a non-separable state.

[0102] Next, an example of the usage method of the culture vessel 1B will be described. Similar to the culture vessel 1 in the above-described Embodiment 1, the culture vessel 1B can be used in a test for evaluating the migratory ability of cells. In the case of the culture vessel 1B, the migratory ability of more types of cells and cells cultured under different culture conditions than the culture vessel 1 in the above-described Embodiment 1 can be evaluated.

[0103] First, an operator prepares a culture vessel 1B in a reference state as shown in FIG. 5. Then, the operator accommodates first target cells in the divided culture region R1 of the culture vessel 1B. Further, the operator accommodates second target cells in the divided culture region R2 of the culture vessel 1B. Further, the operator accommodates third target cells in the divided culture region R3 of the culture vessel 1B. The first target cells, the second target cells, and the third target cells are different types of cells.

[0104] Further, the operator accommodates the same first target cells as those in the divided culture region R1 in the divided culture regions R4, R7, R10, and R13. Further, the operator accommodates culture solutions and / or drugs having different components in the divided culture regions R1, R4, R7, R10, and R13.

[0105] That is, the first target cells accommodated in the divided culture regions R1, R4, R7, R10, and R13 are cultured under different culture conditions.

[0106] In addition, the operator accommodates the same second target cells as those in the divided culture region R2 in the divided culture regions R5, R8, R11, and R14. In addition, the operator accommodates culture solutions and / or drugs with different components in the divided culture regions R2, R5, R8, R11, and R14.

[0107] That is, the second target cells accommodated in the divided culture regions R2, R5, R8, R11, and R14 are cultured under different culture conditions.

[0108] In addition, the operator accommodates the same third target cells as those in the divided culture region R3 in the divided culture regions R6, R9, R12, and R15. In addition, the operator accommodates culture solutions and / or drugs with different components in the divided culture regions R3, R6, R9, R12, and R15.

[0109] That is, the third target cells accommodated in the divided culture regions R3, R6, R9, R12, and R15 are cultured under different culture conditions.

[0110] The operator accommodates the culture vessel 1B in the culture space of a culture apparatus (for example, an incubator). Next, the operator takes out the culture vessel 1B from the culture apparatus (for example, an incubator) after a predetermined time has elapsed since the start of cell culture.

[0111] Before performing the partition removal operation described below, the operator may discard the culture solutions accommodated in the divided culture regions R1 to R15. The necessity of the culture solution discarding operation may be appropriately determined according to the test conditions.

[0112] Next, the operator performs a partition removal operation of removing the partition portion 3B from the culture vessel 1B. In other words, the operator removes the partition portion 3B from the extended region 212B of the bottom surface portion 21B in the culture vessel 1B. When performing the partition removal operation, the operator holds down the main body portion 2B and pulls up the partition portion 3B upward.

[0113] Then, the expansion region 212B is opened, and the divided culture regions R1 to R15 communicate with each other. And the culture vessel 1B is in an expanded state. In the expanded state of the culture vessel 1B, the expansion region 212B is opened, and a second culture space is formed above the expansion region 212B. In this way, the culture space of the culture vessel 1B is expanded.

[0114] In the expanded state of the culture vessel 1B, the operator observes the movements of the first target cells, second target cells, and third target cells cultured in the divided culture regions R1 to R15, and evaluates the migratory ability of the first target cells, second target cells, and third target cells.

[0115] As described above, the culture vessel 1B according to the present embodiment can evaluate the migratory ability of more types of cells and cells cultured under different culture conditions than the culture vessel 1 in the above-described Embodiment 1. Note that the usage method of the culture vessel 1B is not limited to the above-described usage example. The culture vessel 1B may be used in various usage methods according to the conditions of the test to be carried out.

[0116] [Embodiment 3] Next, with reference to FIGS. 8 to 10, a culture vessel and a method for manufacturing the culture vessel according to Embodiment 3 of the present invention will be described.

[0117] As shown in FIG. 1, the culture vessel 1C has a main body portion 2C and a partition portion 3C.

[0118] Also in the culture vessel 1C according to the present embodiment, the size of the culture space changes between the state where the partition portion 3C is supported by the main body portion 2C and the state where the main body portion 2C or the partition portion 3C is removed.

[0119] The configuration of the main body portion 2C is the same as that of the main body portion 2B in the above-described Embodiment 2.

[0120] When viewed as a whole, the partition part 3C is lattice-shaped, similar to the partition part 3B in the above-described Embodiment 2. The partition part 3C is provided in the extended area 212C of the bottom surface part 21C of the main body part 2C. Specifically, the lower surface of the partition part 3C is adsorbed to the extended area 212C. The adsorption force between the extended area 212C and the partition part 3C is such that the culture solution does not flow at least between the divided culture areas R1 to R15.

[0121] In the case of this embodiment, the partition part 3C is divided more finely than the partition part 3B in the above-described Embodiment 2.

[0122] Specifically, the partition part 3C has a plurality of horizontal wall parts 31C, a pair of vertical wall parts 32C1, and a plurality of vertical wall parts 32C2.

[0123] The configuration of the plurality of horizontal wall parts 31C is the same as the configuration of the horizontal wall parts 31B in the above-described Embodiment 2. Each of the plurality of horizontal wall parts 31C has the same length as the length of the bottom surface part 21C in the first direction.

[0124] The pair of vertical wall parts 32C1 are plate-shaped and parallel to the second direction and the third direction. Each of the pair of vertical wall parts 32C1 has the same length as the length of the bottom surface part 21C in the second direction. The pair of vertical wall parts 32C1 are arranged along both end edges of the bottom surface part 21C in the second direction.

[0125] The plurality of vertical wall parts 32C2 are plate-shaped and parallel to the second direction and the third direction. The plurality of vertical wall parts 32C2 have a length that can connect the horizontally adjacent horizontal wall parts 31C in the second direction. That is, the plurality of vertical wall parts 32C2 are shorter than the pair of vertical wall parts 32C1. The plurality of vertical wall parts 32C2 are arranged at equal intervals in the first direction between the pair of vertical wall parts 32C1 and between the horizontally adjacent horizontal wall parts 31C in the second direction.

[0126] In the case of this embodiment, the plurality of horizontal wall parts 31C and the pair of vertical wall parts 32C1 are integrally formed in a non-separable state. Also, the plurality of vertical wall parts 32C2 are configured as separate members from the plurality of horizontal wall parts 31C and the pair of vertical wall parts 32C1.

[0127] Further, the plurality of horizontal wall portions 31C and the pair of vertical wall portions 32C1 may be included in the main body portion 2C. That is, the plurality of horizontal wall portions 31C and the pair of vertical wall portions 32C1 may be integrally formed with the main body portion 2C in a state where they cannot be separated from the main body portion 2C.

[0128] Similar to the culture vessel 1B in the above-described Embodiment 2, the culture vessel 1C as described above can evaluate the migration ability of many types of cells and cells cultured under different culture conditions.

[0129] In particular, in the case of the present embodiment, by changing the timing of removing the plurality of vertical wall portions 32C2 in the partition portion 3C from the main body portion 2C, the timing of communicating the divided culture regions (for example, divided culture regions R1 to R3) adjacent to each other in the first direction can be changed.

[0130] Specifically, after removing the vertical wall portion 32C2 disposed between the divided culture region R1 and the divided culture region R2 from the main body portion 2C, the vertical wall portion 32C2 disposed between the divided culture region R2 and the divided culture region R3 is removed from the main body portion 2C. According to such a procedure, the timing at which the divided culture region R1 and the divided culture region R2 communicate and the timing at which the divided culture region R2 and the divided culture region R3 communicate can be made different.

[0131] Further, the timing of removing the horizontal wall portion 31C and the vertical wall portion 32C1 in the partition portion 3C from the main body portion 2C and the timing of removing the vertical wall portion 32C2 from the main body portion 2C can also be made different. According to the culture vessel 1C according to such a present embodiment, a test with a high degree of freedom can be carried out.

[0132] [Embodiment 4] Next, with reference to FIGS. 11 and 12, a culture vessel and a method for manufacturing the culture vessel according to Embodiment 4 of the present invention will be described. FIG. 11 is a plan view of a culture vessel 1D according to Embodiment 4. For reference, FIG. 11 is a plan view showing the reference state of the culture vessel 1D. FIG. 12 is a plan view of a main body portion 2D of the culture vessel 1D according to Embodiment 4. In other words, FIG. 12 is a plan view of the culture vessel 1D in an expanded state.

[0133] As shown in FIG. 11, the culture vessel 1D has a main body portion 2D and a partition portion 3D.

[0134] In the culture vessel 1D according to the present embodiment, the size of the culture space also changes between the state where the partition portion 3D is supported by the main body portion 2D and the state where the main body portion 2D or the partition portion 3D is removed.

[0135] The shape of the main body portion 2D is the same as that of the main body portion 2B in Embodiment 2 described above. However, in the main body portion 2D, the shapes of the culture region 211D and the expansion region 212D on the bottom surface portion 21D are different from the shapes of the culture region 211B and the expansion region 212B in Embodiment 2 described above.

[0136] Further, the surface of the culture region 211B on the bottom surface portion 21D is coated with a scaffold material that promotes cell adhesion. On the other hand, the surface of the expansion region 212B on the bottom surface portion 21D is not coated with a scaffold material. The scaffold material is not particularly limited, and for example, collagen, fibronectin, laminin, gelatin, vitronectin, poly-L-lysine, and adhesion peptides can be used.

[0137] The culture region 211D is the region not marked with oblique grids in FIG. 12. The culture region 211D has a shape along the final shape (in other words, the shape after culturing) of the target cells. In other words, the target cells finally grow into a shape along the culture region 211D.

[0138] On the one hand, the expansion region 212D is the region marked with a diagonal grid in FIG. 12. The expansion region 212D is a region surrounding the culture region 211D. In other words, the shape of the culture region 211D is defined by the expansion region 212D.

[0139] In the case of this embodiment, the culture region 211D and the expansion region 212D are provided on the same plane. However, the culture region 211D may be constituted by a concave portion. That is, the culture region 211D may be provided below the expansion region 212D.

[0140] The partition portion 3D has a shape that overlaps with the expansion region 212B in a plan view. The partition portion 3D is provided in the expansion region 212D of the bottom surface portion 21D. Specifically, the lower surface of the partition portion 3D is adsorbed to the expansion region 212D.

[0141] Next, an example of the usage method of the culture vessel 1D will be described. According to the culture vessel 1D, cells that can follow the shape of the culture region 211D can be cultured. The culture vessel 1D can be used when culturing cells that have the property of specifically adhering to the surface coated with the scaffold material (scaffold-dependent adhesiveness). The following description is also an example of a method for culturing cells implemented using the culture vessel 1D.

[0142] By the way, when the target cells grow to the final shape in the state where the partition portion 3D exists, the target cells may adhere to the inner surface of the partition portion 3D. In this state, when an operator tries to take out the target cells from the culture vessel 1D, the target cells may be damaged.

[0143] Moreover, if the culture of the target cells is continued in the space surrounded by the partition portion 3D, the target cells may be damaged due to reasons such as depletion of nutrients and oxygen, and excessive density of the target cells.

[0144] Therefore, in the case of this embodiment, the operator removes the partition part 3D from the main body part 2D during cell culture. That is, the expansion region 212D is opened. The timing of removing the partition part 3D from the main body part 2D is preferably before the target cells grow to the final shape.

[0145] In addition, the partition part 3D may be removed from the main body part 2D before the start of cell culture (for example, before seeding the cells), or the partition part 3D may be removed from the main body part 2D after the start of cell culture (for example, after seeding the cells or after culturing for a certain period after seeding the cells).

[0146] When the partition part 3D is removed from the main body part 2D before the start of cell culture (for example, before seeding the cells), such a process corresponds to an example of the fourth step in the cell culture method. The fourth step in the cell culture method is a step of removing the partition part 3D from the main body part 2D to open the expansion region 212D.

[0147] In this case, in the state after opening the expansion region 212D (that is, the expanded state of the culture vessel 1D), cell culture is started on the culture surface. Such a process corresponds to an example of the fifth step of the cell culture method.

[0148] By removing the partition part 3D from the main body part 2D before the target cells grow to the final shape, it is possible to suppress the target cells from adhering to the inner surface of the partition part 3D. As a result, when the operator takes out the target cells from the culture vessel 1D, it is possible to suppress the target cells from being damaged.

[0149] In addition, the culture vessel 1D according to this embodiment can also be used in a test for evaluating the migratory ability of cells, similar to the culture vessels 1, 1B, and 1C according to the above-described Embodiments 1 to 3.

Industrial Applicability

[0150] The culture vessel according to the present invention can be applied to the culture of various cells.

Explanation of Reference Numerals

[0151] 1, 1B, 1C, 1D culture vessels 2, 2B, 2C, 2D main body parts 21, 21B, 21C, 21D bottom face parts 211, 211B, 211D culture areas Ra, Rb divided culture areas R1, R2, R3, R4, R5, R6, R7, R8 divided culture areas R9, R10, R11, R12, R13, R14, R15 divided culture areas 212, 212B, 212C, 212D expansion areas 22 peripheral wall parts 23 scale 3, 3B, 3C, 3D partition parts 31B, 31C horizontal wall parts 32B, 32C1, 32C2 vertical wall parts

Claims

1. A main body having a culture area and an expansion area on a culture surface, and a partition wall covering the expansion area from above, wherein when the partition wall is removed from the expansion area, the expansion area is opened, a culture vessel.

2. The partition wall is adsorbed to the expansion area, the culture vessel according to Claim 1.

3. The culture area is divided into a plurality of divided culture areas by the expansion area, the culture vessel according to Claim 1.

4. The culture area is divided into at least a first section and a second section by the expansion area, and the first section and the second section are used for culturing different types of cells, the culture vessel according to Claim 1.

5. The partition wall separates the culture spaces provided above adjacent culture areas, and when the partition wall is removed from the expansion area, the expansion area is opened and the adjacent culture spaces communicate with each other, the culture vessel according to Claim 1.

6. The main body further includes a peripheral wall surrounding the culture surface, the culture vessel according to Claim 1.

7. The culture area is an area surrounded by the expansion area, the culture vessel according to Claim 1.

8. The expansion area has a scale for measuring dimensions, the culture vessel according to Claim 1.

9. The oxygen permeability of the culture surface at 23°C is 7,500 cm ３ / (m ２ ·24 h·atm) or more and 150,000 cm ３ / (m ２ ·24 h·atm) or less, The culture vessel according to Claim 1.

10. The surface of the culture area is coated with a scaffold material that promotes cell adhesion, and the surface of the expansion area is not coated with the scaffold material, the culture vessel according to Claim 1.

11. The scaffold material contains at least one selected from the group consisting of collagen, fibronectin, laminin, gelatin, vitronectin, poly-L-lysine, and adhesion peptides, the culture vessel according to Claim 10.

12. The main body includes a polymer having a structural unit derived from 4-methyl-1-pentene, the culture vessel according to Claim 1.

13. The partition wall includes a polymer having a structural unit derived from 4-methyl-1-pentene, the culture vessel according to Claim 1.

14. A step of providing a main body having a culture area and an expansion area on a culture surface, a step of providing a partition wall at a position facing the expansion area, and a step of bringing the partition wall into close contact with the expansion area, including, a method for manufacturing a culture vessel.

15. Using the culture vessel according to any one of Claims 1 to 11, a method for culturing cells.

16. A step of starting culturing cells in the culturing region of the culturing surface; A step of removing the partition portion to open the expansion region; A step of further continuing culturing cells on the culturing surface with the expansion region opened; The method for culturing cells according to claim 15, comprising the above steps.

17. The method for culturing cells according to claim 16, further comprising a step of administering a drug to the cells.

18. A step of removing the partition portion to open the expansion region; A step of starting culturing cells on the culturing surface with the expansion region opened; The method for culturing cells according to claim 15, comprising the above steps.

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