Cell culture device

Abstract

To easily maintain airtightness in a cell culture device in which a lid is opened and closed.SOLUTION: A cell culture device 1 comprises: a culture container 2 comprising a plurality of reservoirs 21 having openings; a lid 3 that is removably attached to the culture container 2 and covers the openings of the plurality of reservoirs 21; a pneumatic piping 4 connected to one of the culture container 2 and the lid 3; and a rubber sheet 5 disposed between the culture container 2 and the lid 3 to ensure airtightness between the culture container 2 and the lid 3. A linear seal protrusion 60 that surrounds the openings of the plurality of reservoirs 21 and protrudes toward the rubber sheet 5 is provided on one of the culture container 2 and the lid 3.SELECTED DRAWING: Figure 2

Description

Technical Field

[0001] The present invention relates to a cell culture device.

Background Art

[0002] The following Patent Document 1 discloses a cell culture device. This cell culture device includes a storage tank and a pressure pump. The storage tank includes a container-shaped tank body and a lid. The lid can open and close the opening of the tank body and can be hermetically closed. Specifically, the lid can hermetically close the upper openings of the first culture solution storage chamber and the second culture solution storage chamber of the tank body, respectively. The lid is in contact with the upper surface of the tank body through packing provided so as to surround each of the upper openings.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] When culturing cells using the above cell culture device, periodic replacement of the culture solution is required. When replacing the culture solution, the lid of the cell culture device is opened and closed. At this time, there is a risk that the sealing material such as the above packing may fall off. In addition, a large number of conventional sealing materials are arranged so as to surround the upper openings of each of the plurality of storage parts, the number of parts is large, and the assembly is complicated. Further, if the dimensional error between the culture container and the lid is large, there may be a case where the dimensional error cannot be absorbed by the elastic deformation of each sealing material, and a problem may occur in maintaining airtightness.

[0005] The present invention has been made in view of the above problems, and an object thereof is to facilitate airtightness maintenance in a cell culture device in which the lid is opened and closed.

Means for Solving the Problems

[0006] A cell culture apparatus according to one aspect of the present invention comprises a culture vessel having a plurality of storage compartments having openings, a lid detachably attached to the culture vessel and covering the openings of the plurality of storage compartments, pneumatic piping connected to either the culture vessel or the lid, and a rubber sheet disposed between the culture vessel and the lid to ensure airtightness between the culture vessel and the lid, wherein the culture vessel or the lid is provided with linear sealing protrusions that surround the openings of the plurality of storage compartments and project toward the rubber sheet.

[0007] Furthermore, in a cell culture apparatus according to one aspect of the present invention, the sealing projection may be provided on the culture vessel.

[0008] Furthermore, in a cell culture apparatus according to one aspect of the present invention, the lid may have a smooth surface to which the rubber sheet adheres.

[0009] Furthermore, in a cell culture apparatus according to one aspect of the present invention, the lid may have a plurality of positioning protrusions for positioning the rubber sheet on the smooth surface.

[0010] Furthermore, in a cell culture apparatus according to one aspect of the present invention, the plurality of positioning protrusions may be provided on both sides of the rubber sheet in the short direction and on one side of the rubber sheet in the longitudinal direction.

[0011] Furthermore, in a cell culture apparatus according to one aspect of the present invention, the culture vessel may be provided with a plurality of positioning recesses into which the plurality of positioning protrusions are inserted.

[0012] Furthermore, in a cell culture apparatus according to one aspect of the present invention, the pneumatic piping may be connected to the lid.

[0013] Furthermore, in a cell culture apparatus according to one aspect of the present invention, the lid may be provided with a plurality of grooves that communicate with the pneumatic piping, and the rubber sheet may be provided with a plurality of communication holes that connect the plurality of grooves with the plurality of storage sections.

[0014] Also, in the cell culture device according to one aspect of the present invention, the seal protrusion may be provided along the opening edge of each of the plurality of storage parts.

[0015] Also, in the cell culture device according to one aspect of the present invention, at least a part of the seal protrusions provided along the opening edges of the adjacent storage parts among the plurality of storage parts may be shared.

[0016] Also, in the cell culture device according to one aspect of the present invention, the ratio of the height / width of the seal protrusion may be within the range of 0.5 to 10.

[0017] Also, in the cell culture device according to one aspect of the present invention, the rubber sheet may be made of silicone rubber.

[0018] Also, in the cell culture device according to one aspect of the present invention, the durometer hardness of the rubber sheet may be 50 or less.

Advantages of the Invention

[0019] According to one aspect of the present invention described above, it is possible to easily maintain airtightness in a cell culture device for opening and closing a lid.

Brief Description of the Drawings

[0020] [Figure 1] It is a perspective view showing a cell culture device according to the first embodiment. [Figure 2] It is an exploded perspective view of the cell culture device shown in FIG. 1. [Figure 3] It is an exploded perspective view of the lid side of the cell culture device shown in FIG. 1. [Figure 4] It is an enlarged view of the region A shown in FIG. 2. [Figure 5] It is a cross-sectional view taken along the arrow V-V of the cell culture device shown in FIG. 1. [Figure 6] It is a perspective view of a cell culture device according to the second embodiment. [Figure 7]It is an exploded perspective view of the cell culture device shown in FIG. 6. [Figure 8] It is a plan view of the culture vessel included in the cell culture device shown in FIG. 7. [Figure 9] It is a cross-sectional view taken along arrow IX-IX shown in FIG. 8.

Embodiments for Carrying Out the Invention

[0021] Hereinafter, each embodiment of the present invention will be described with reference to the drawings. Note that "~" indicating a numerical range means including the numerical values described before and after it as the lower limit value and the upper limit value.

[0022] [ (First Embodiment) FIG. 1 is a perspective view of a cell culture device 1 according to the first embodiment. FIG. 2 is an exploded perspective view of the cell culture device 1 shown in FIG. 1. FIG. 3 is an exploded perspective view of the lid side of the cell culture device 1 shown in FIG. 1. FIG. 4 is an enlarged view of region A shown in FIG. 2. FIG. 5 is a cross-sectional view taken along arrow V-V shown in FIG. 1. As shown in FIGS. 1 and 2, the cell culture device 1 includes a culture vessel 2, a lid 3, a pneumatic pipe 4, and a rubber sheet 5.

[0023] In the following description, an XYZ orthogonal coordinate system may be set, and the positional relationship of each component may be described while referring to this XYZ orthogonal coordinate system. The X-axis direction of the first embodiment is the first horizontal direction and is the short side direction of the cell culture device 1. The Y-axis direction of the first embodiment is the second horizontal direction orthogonal to the first horizontal direction and is the longitudinal direction of the cell culture device 1. The Z-axis direction of the first embodiment is the vertical direction orthogonal to the first horizontal direction and the second horizontal direction and is the height direction (thickness direction) of the cell culture device 1.

[0024] As shown in Figure 2, the culture vessel 2 is equipped with a plurality of storage compartments 21. The storage compartments 21 store liquid, such as a culture medium for culturing cells. The storage compartments 21 open to the upper surface 20A of the culture vessel 2. The storage compartments 21 have a roughly rectangular shape in plan view, with the X-axis direction being the short side and the Y-axis direction being the long side. The storage compartments 21 are arranged in a 4x4 configuration in the XY plane. However, the shape, number, and arrangement of the storage compartments 21 are not limited to this.

[0025] As shown in Figure 1, the culture vessel 2 comprises an upper plate 2a and a bottom plate 2b. The upper plate 2a can be manufactured, for example, by cutting polycarbonate. The bottom plate 2b can be manufactured, for example, by cutting stainless steel. The upper plate 2a and bottom plate 2b can also be manufactured by injection molding of a thermoplastic resin such as polystyrene. On both sides of the bottom plate 2b in the X-axis direction, there are first fixing parts 6 for fixing the upper plate 2a to the upper part of the bottom plate 2b. The first fixing parts 6 have a substantially C-shape extending in the Y-axis direction, and both ends are fixed to the bottom plate 2b so as to be rotatable around an axis extending in the Y-axis direction.

[0026] On both sides of the upper plate 2a in the X-axis direction, there are first locking parts 8 that engage with the first fixing part 6 in the Z-axis direction. The upper plate 2a and the bottom plate 2b are fixed together when the first fixing part 6 rotates around the Y-axis and engages with the first locking part 8 located above it. The first locking parts 8 are provided in pairs, spaced apart in the Y-axis direction. A gap 9 is formed between the pair of first locking parts 8, allowing a fingertip to be inserted into the lower edge of the lid 3. This facilitates the removal of the lid 3 and the transport of the cell culture apparatus 1.

[0027] Furthermore, second fixing parts 7 are provided on both sides of the bottom plate 2b in the X-axis direction to secure the lid 3 to the upper part of the upper plate 2a. The second fixing parts 7 have a roughly C-shape extending in the Y-axis direction, and both ends are fixed to the bottom plate 2b so as to be rotatable around an axis extending in the Y-axis direction. The second fixing parts 7 are slightly larger than the first fixing parts 6.

[0028] On both sides of the lid 3 in the X-axis direction, there are second locking parts 10 that engage with the second fixing part 7 in the Z-axis direction. The lid 3 and the bottom plate 2b (culture container 2) are fixed together when the second fixing part 7 rotates around the Y-axis and engages with the second locking part 10 located above it. When removing the lid 3 from the culture container 2, the pair of second fixing parts 7 are rotated to open on both sides in the X-axis direction.

[0029] As shown in Figure 5, a flow channel plate 2c and a transparent plate 2d are sandwiched between the upper plate 2a and the bottom plate 2b. The flow channel plate 2c can be manufactured, for example, by replica molding of silicone resin or injection molding of polystyrene. The flow channel plate 2c is positioned below the upper plate 2a, and the transparent plate 2d is positioned below the flow channel plate 2c. The flow channel plate 2c has through holes 14, an outlet channel 15, and an introduction channel 16 formed therein. The transparent plate 2d is a transparent plate material such as a glass plate, acrylic plate, polystyrene plate, or silicone resin plate.

[0030] The through-hole 14 is positioned in the upper plate 2a at a location corresponding to the bottom surface of the storage section 21. The lower end of the through-hole 14 is closed by the transparent plate 2d. The bottom plate 2b has a window section 13 (opening) formed in a location corresponding to the through-hole 14. This allows observation of the inside of the storage section 21 from the bottom side of the cell culture apparatus 1 through the window section 13, the transparent plate 2d, and the through-hole 14.

[0031] The discharge channel 15 is located near the through hole 14 and communicates with the bottom surface of the storage section 21. On the other hand, the introduction channel 16 communicates with a stepped portion that is higher than the bottom surface of the storage section 21. As shown in Figure 2, the discharge channel 15 provided in any one of the storage sections 21a to 21d arranged in the X-axis direction communicates with the introduction channel 16 of the other storage sections 21. For example, two sets of storage sections 21a to 21d may be connected, or all of the storage sections 21a to 21d may be connected in a complete loop.

[0032] The pneumatic piping 4 includes pneumatic pipes 4a to 4d that apply air pressure to the storage sections 21a to 21d. By selectively applying air pressure from the pneumatic pipes 4a to 4d to the storage sections 21a to 21d, an air pressure difference is created between the storage sections 21, allowing the liquid stored in one storage section 21 to be supplied to the other storage sections 21, and thus the liquid to be circulated (perfused). In addition, near the discharge channel 15, the channel cross-sectional area is 0.1 [mm²] 2 A thin Laplace valve 151 is provided below, and the introduction channel 16 is connected to an opening 70 that is one level higher than the bottom surface of the storage section 21, so that backflow of liquid can be prevented when air pressure is applied.

[0033] As shown in Figure 1, the lid 3 comprises a lid body 3a and a cover 3b. The cover 3b is made of a transparent plate material such as glass or acrylic. The cover 3b is fixed to the upper part of the lid body 3a via a plurality of bolts 11. The bolts 11 are screwed into the lid body 3a through one of a plurality of through holes 12 formed in the cover 3b. In this example, the number of through holes 12 is greater than the number of bolts 11, but the number may be equal. The lower end of the through holes 12 communicates with a bottomed screw hole (not shown) formed in the lid body 3a.

[0034] As shown in Figure 3, a connection hole 30a is formed on one longitudinal side (+Y side) of the lid body 3a, to which the pneumatic piping 4 is connected. The lid body 3a is provided with a plurality of grooves 31 that communicate with the pneumatic piping 4. The grooves 31 open into the smooth surface 30B on the lower surface of the lid body 3a. The upper surface 30A side of the grooves 31 of the lid body 3a is closed by the cover 3b. In order to maintain airtightness, a rubber sheet 3c similar to the rubber sheet 5 described later is sandwiched between the lid body 3a and the cover 3b, as shown in Figure 5.

[0035] As shown in Figure 3, the groove 31 has a roughly rectangular shape when viewed from below, with the X-axis direction as the short side and the Y-axis direction as the long side. The groove 31 is arranged in 4 rows x 4 columns in the XY plane, the same number as the storage section 21. The shape, number, and arrangement of the groove 31 are not limited to those corresponding to the shape, number, and arrangement of the storage section 21. Among the multiple groove 31, adjacent groove 31 in the longitudinal direction (Y-axis direction) of the lid body 3a are connected to each other by through holes 32 formed in the lid body 3a. The through holes 32 penetrate the side walls of adjacent groove 31 in the Y-axis direction. In other words, the pneumatic piping 4a to 4d can be compressed into each row of grooves 31a to 31d.

[0036] As shown in Figure 2, the rubber sheet 5 is interposed between the culture container 2 and the lid 3 to ensure airtightness between the culture container 2 and the lid 3. The rubber sheet 5 has a rectangular shape in plan view, with the X-axis direction being the short side and the Y-axis direction being the long side. The upper surface 50A and lower surface 50B of the rubber sheet 5 are both smooth surfaces. The upper surface 50A of the rubber sheet 5 adheres tightly to the smooth surface 30B of the lid 3 shown in Figure 3. In other words, the rubber sheet 5 adheres tightly to the lid 3 side due to van der Waals forces.

[0037] The rubber sheet 5 is formed from, for example, silicone rubber. However, the rubber sheet 5 may be made of rubber other than silicone rubber. The durometer hardness of the rubber sheet 5 should preferably be 50 or less. Here, the durometer hardness of the rubber sheet 5 conforms to the durometer type A of JIS standard K6253. The thickness T of the rubber sheet 5 (see Figure 2) should preferably be greater than the height H of the sealing protrusion 60 (see Figure 4), which will be described later. On the other hand, if the thickness T of the rubber sheet 5 is too large, it will impair the operability of the cell culture apparatus. Specifically, the thickness T of the rubber sheet 5 is preferably in the range of 1 to 10 [mm].

[0038] The rubber sheet 5 is provided with multiple communication holes 51 that connect multiple grooves 31 provided in the lid 3 and multiple storage sections 21 provided in the culture container 2. The communication holes 51 penetrate the rubber sheet 5 in the Z-axis direction. The communication holes 51 have a substantially rectangular shape in plan view, with the X-axis direction as the short side and the Y-axis direction as the long side. The communication holes 51 are arranged in a 4x4 configuration in the XY plane, the same number as the grooves 31 and storage sections 21. The shape, number, and arrangement of the communication holes 51 are not limited to those described above, as long as they correspond to the shape, number, and arrangement of the grooves 31 and storage sections 21.

[0039] As shown in Figure 3, the lid 3 has a plurality of positioning protrusions 33 that position the rubber sheet 5 on the smooth surface 30B. This allows the grooves 31a to 31d and the communication holes 51a to 51d to be aligned. The positioning protrusions 33 are provided in a total of three locations: on both sides in the short direction (X-axis direction) of the rubber sheet 5 and on one side in the longitudinal direction (+Y side) of the rubber sheet 5. When placing the rubber sheet 5 on the lid 3, the positioning protrusions 33 can be positioned adjacent to the outer shape of the rubber sheet 5 on both sides in the short direction (X-axis direction) and one side in the longitudinal direction (+Y side), thereby determining the position of the rubber sheet 5 on the XY plane. The positioning protrusions 33 protrude downward from the smooth surface 30B. The length from the smooth surface 30B to the lower end of the positioning protrusions 33 is greater than the thickness T of the rubber sheet 5.

[0040] As shown in Figure 2, the culture vessel 2 is equipped with multiple positioning recesses 22 into which multiple positioning protrusions 33 are inserted. There are a total of three positioning recesses 22, one on each side in the short direction (X-axis direction) of the culture vessel 2 and one side in the long direction (+Y side) of the culture vessel 2. By inserting the three positioning protrusions 33 provided on the lower part of the lid 3 into the three positioning recesses 22 provided on the upper part of the culture vessel 2, it is possible to prevent the lid 3 from being assembled incorrectly on the culture vessel 2 (for example, assembly with the direction of the pneumatic piping 4 in the Y-axis direction reversed).

[0041] As shown in Figure 4, the culture vessel 2 is provided with linear sealing protrusions 60 that surround the openings of multiple storage sections 21 and project from the upper surface 20A toward the rubber sheet 5. The sealing protrusions 60 are provided along the opening edges of each of the multiple storage sections 21. The sealing protrusions 60 have a roughly rectangular ring shape in plan view, slightly larger than the storage section 21, with the X-axis direction as the short side and the Y-axis direction as the long side. Specifically, the sealing protrusions 60 include a pair of short sides 61 extending parallel to the X-axis direction, a pair of long sides 62 extending parallel to the Y-axis direction, and four corners 63 connecting the ends of the pair of short sides 61 and the ends of the pair of long sides 62. In plan view, the corners 63 are curved in an arc shape.

[0042] The tip of the sealing projection 60 is formed in a planar shape. Alternatively, the tip of the sealing projection 60 may be formed in a roughly semicircular shape in a cross-sectional view that is convex upwards. At least a portion of the sealing projections 60 provided along the opening edges of adjacent storage sections 21 are common to each other. Specifically, in Figure 4, the sealing projection 60 of the storage section 21d located at the corner of the culture vessel 2 has its short side 61 on the +Y side and its long side 62 on the -X side common to the short side 61 or long side 62 of the sealing projection 60 of the adjacent storage section 21.

[0043] The height / width ratio of the sealing protrusion 60 should be within the range of 0.5 to 10. Specifically, if the width W of the sealing protrusion 60 is 1.0 [mm], the height H of the sealing protrusion 60 should be within the range of 0.5 to 10 [mm]. Furthermore, the height / width ratio of the sealing protrusion 60 should be the same even in the common parts. This ensures that the amount of elastic deformation of the rubber sheet 5 is uniform in each storage section 21, thereby improving airtightness. In addition, the culture vessel 2 has a roughly rhomboid space 64 formed by the corners 63 of the sealing protrusions 60 of the four adjacent storage sections 21. The space 64 has a volume such that the amount of elastic deformation of the rubber sheet 5 at the four corners 63 is the same in the inner and outer regions of the four adjacent annular sealing protrusions 60.

[0044] In the cell culture apparatus 1 with the above configuration, the culture vessel 2, which has multiple storage compartments 21, is equipped with a lid 3 that covers the culture vessel 2, and a rubber sheet 5 is placed between the culture vessel 2 and the lid 3 to ensure airtightness of the multiple storage compartments 21 simultaneously. At this time, by providing linear sealing protrusions 60 on the upper surface of the culture vessel 2, the rubber sheet 5 is elastically deformed, and airtightness can be ensured in the area along the sealing protrusions 60. With this rubber sheet 5, airtightness of multiple storage compartments 21 can be ensured with a single component, making assembly easy, and by adjusting the thickness T of the rubber sheet 5 and the height H of the sealing protrusions 60, dimensional errors of the culture vessel 2 and the lid 3 can be absorbed and airtightness can be easily ensured.

[0045] As described above, the cell culture apparatus 1 according to this embodiment comprises a culture container 2 having a plurality of storage sections 21 with openings, a lid 3 detachably attached to the culture container 2 and covering the openings of the plurality of storage sections 21, a pneumatic pipe 4 connected to either the culture container 2 or the lid 3 (in this embodiment, the lid 3), and a rubber sheet 5 positioned between the culture container 2 and the lid 3 to ensure airtightness between the culture container 2 and the lid 3. Linear sealing protrusions 60 are provided on either the culture container 2 or the lid 3, surrounding the openings of the plurality of storage sections 21 and projecting toward the rubber sheet 5. This configuration makes it easy to maintain airtightness in the cell culture apparatus 1 when opening and closing the lid 3.

[0046] Furthermore, in this embodiment, the sealing projection 60 is provided on the culture container 2, and the lid 3 has a smooth surface 30B to which the rubber sheet 5 adheres. With this configuration, when the lid 3 is opened, the rubber sheet 5 adheres to the lid 3 side rather than the culture container 2 side. This allows the rubber sheet 5 to be removed together with the lid 3, and the adhesion with the lid 3 prevents the rubber sheet 5 from falling off. However, when replenishing the liquid in the storage section 21 after removing the lid 3, if the rubber sheet 5 is in close contact with the upper surface 20A of the culture container 2, it is undesirable because if the liquid seeps into the gap between the rubber sheet 5 and the culture container 2, it could cause contamination. Also, the liquid that seeps into the gap may deteriorate and affect cell culture. With this configuration, the above concerns can be eliminated.

[0047] Furthermore, in this embodiment, the lid 3 has a plurality of positioning protrusions 33 for positioning the rubber sheet 5 on the smooth surface 30B. This configuration makes it easy to position the rubber sheet 5 on the smooth surface 30B of the lid 3.

[0048] Furthermore, in this embodiment, the multiple positioning protrusions 33 are provided on both sides of the rubber sheet 5 in the short direction and on one side of the rubber sheet 5 in the longitudinal direction. The culture container 2 also has multiple positioning recesses 22 into which the multiple positioning protrusions 33 are inserted. With this configuration, the positioning structure of the rubber sheet 5 can be used to prevent the lid 3 from being assembled incorrectly on the culture container 2 (for example, assembly with the direction of the pneumatic piping 4 in the Y-axis direction reversed). This makes it possible to apply pneumatic pressure to the storage section 21 as designed under predetermined conditions.

[0049] In this embodiment, the pneumatic piping 4 is connected to the lid 3. The lid 3 is provided with a plurality of grooves 31 that communicate with the pneumatic piping 4, and the rubber sheet 5 is provided with a plurality of communication holes 51 that connect the plurality of grooves 31 to the plurality of storage sections 21. With this configuration, since the pneumatic piping 4 is positioned higher than the storage section 21, even if air bubbles are mixed in during the circulation (perfusion) of the culture medium and the liquid in the storage section 21 becomes foamy, it is possible to suppress the entry of such bubbles into the pneumatic piping 4. This makes it possible to apply pneumatic pressure to the storage section 21 as designed under predetermined conditions. If the storage section 21 is made deeper, the pneumatic piping 4 may be connected to the culture container 2.

[0050] Furthermore, in this embodiment, the sealing projections 60 are provided along the opening edges of each of the multiple storage sections 21. In addition, at least a portion of the sealing projections 60 provided along the opening edges of adjacent storage sections 21 are common to all of the multiple storage sections 21. This configuration allows for space-saving arrangement of the storage sections 21 in the culture vessel 2 while ensuring uniform airtightness in each storage section 21.

[0051] Furthermore, in this embodiment, the height / width ratio of the sealing projection 60 is within the range of 0.5 to 10. This configuration ensures airtightness and prevents the rubber sheet 5 from falling off, while also ensuring sufficient physical strength of the sealing projection. In particular, if the height H of the sealing projection 60 is 0.3 [mm] or more, it can absorb dimensional errors of the lid 3 and culture container 2, thereby ensuring even greater airtightness. Moreover, if the width W of the sealing projection 60 is 2.0 [mm] or less, it ensures airtightness and prevents the rubber sheet from falling off. Additionally, if the rubber sheet 5 is made of silicone rubber and its durometer hardness is 50 or less, it further enhances airtightness and prevents the rubber sheet from falling off.

[0052] (Second Embodiment) Next, a second embodiment of the present invention will be described. In the following description, components identical or equivalent to those in the above-described embodiment will be denoted by the same reference numerals, and their descriptions will be simplified or omitted.

[0053] Figure 6 is a perspective view of the cell culture apparatus 1 according to the second embodiment. Figure 7 is an exploded perspective view of the cell culture apparatus 1 shown in Figure 6. Figure 8 is a plan view of the culture vessel 2 included in the cell culture apparatus 1 shown in Figure 7. Figure 9 is a cross-sectional view taken along the line IX-IX shown in Figure 8. The cell culture apparatus 1 of the second embodiment differs from the above embodiment in that a hydrogel chamber 110 is provided between a pair of storage sections 21a and 21b of the culture vessel 2.

[0054] In the following explanation, an XYZ Cartesian coordinate system will be established, and the positional relationships of each component will be described with reference to this XYZ Cartesian coordinate system. The X-axis direction is the first horizontal direction in which the reservoir 21a, hydrogel chamber 110, and reservoir 21b are aligned. The Y-axis direction is the second horizontal direction perpendicular to the X-axis direction. The Z-axis direction is the vertical direction perpendicular to both the X-axis and Y-axis directions.

[0055] In Figure 6, the external shapes of the culture container 2, lid 3, and rubber sheet 5 are shown with dashed lines to improve the visibility of the internal structure of the cell culture apparatus 1, allowing the internal structure to be seen through them. The culture container 2 and lid 3 are preferably made of a highly translucent transparent material for observing cells, but they may also be made of a material with low or no translucency. In that case, a glass window or the like may be provided in a position where the cells can be seen. The cell culture apparatus 1 of the second embodiment can be manufactured, for example, by injection molding and heat sealing of polystyrene.

[0056] As shown in Figure 6, the hydrogel chamber 110 has a flattened internal space. The height (dimension in the Z-axis direction) of the internal space of the hydrogel 101 is, for example, in the range of 100 [μm] to 500 [μm]. The internal space of the hydrogel chamber 110 is filled with hydrogel 101. The hydrogel 101 has a three-dimensional network structure of polymers that retains moisture, embeds or adheres cells, and allows for cell culture.

[0057] The hydrogel chamber 110 is formed in a hexagonal shape in plan view. While the plan view shape of the hydrogel chamber 110 is not particularly limited, the length adjacent to the reservoirs 21a and 21b (the dimension in the Y-axis direction) should preferably be greater than the width (the dimension in the X-axis direction). The Y-axis dimension of the hydrogel chamber 110 is, for example, within the range of 1 mm to 20 mm. The X-axis dimension of the hydrogel chamber 110 is, for example, within the range of 0.5 mm to 10 mm.

[0058] The hydrogel chamber 110 has a first surface communicating with the storage section 21a and a second surface communicating with the storage section 21b. The first and second surfaces face each other in the X-axis direction and extend parallel to each other in the Y-axis direction. Both ends of the hydrogel chamber 110 in the Y-axis direction communicate with the introduction holes 114. A hydrogel storage tank 116 for introducing the hydrogel 101 is formed directly above the hydrogel chamber 110.

[0059] The inlet holes 114 connect both ends of the hydrogel chamber 110 in the Y-axis direction to the bottom surface of the hydrogel storage tank 116. The two inlet holes 114 make it easier to remove air from the hydrogel chamber 110 and to fill the hydrogel chamber 110 with hydrogel 101 from the hydrogel storage tank 116.

[0060] The storage section 21a stores the culture medium 102. The storage section 21a is formed in the shape of an elongated slit or ellipse extending in the Y-axis direction in a plan view. The plan view shape of the storage section 21a is not particularly limited. The storage section 21a should preferably have a larger internal space (volume) than the hydrogel chamber 110. The side of the bottom of the storage section 21a on the hydrogel chamber 110 side (+X side) is in communication with the hydrogel chamber 110.

[0061] The storage section 21b stores the culture medium 102. The storage section 21b is formed in a long slit or elliptical shape extending in the Y-axis direction in a plan view. The planar shape of the storage section 21b is not particularly limited. It is preferable that the internal space (volume) of the storage section 21b is larger than that of the hydrogel chamber 110. The side of the bottom of the storage section 21b on the hydrogel chamber 110 side (-X side) is in communication with the hydrogel chamber 110.

[0062] The culture vessel 2 is equipped with column rows 122 and 132 as a mechanism to retain the hydrogel 101 in the hydrogel chamber 110 even when a pressure difference is created between the storage sections 21a and 21b. Column row 122 forms a micro-sized fluid channel connecting the hydrogel chamber 110 and the storage section 21a. Column row 132 also forms a micro-sized fluid channel connecting the hydrogel chamber 110 and the storage section 21b.

[0063] The smooth surface 30B (bottom surface) of the lid 3 has upward-facing concave grooves 31a and 31b that communicate with the storage sections 21a and 21b. The upper part of the hydrogel storage tank 116 is closed by the smooth surface 30B of the lid 3. Pneumatic pipes 4a and 4b are connected to the lid 3 so as to communicate with the storage sections 21a and 21b. The direction in which the pneumatic pipes 4a and 4b are connected to the lid 3 is not limited to the Y-axis direction, but may also be the X-axis direction or the Z-axis direction.

[0064] As shown in Figures 6 and 7, the rubber sheet 5 is interposed between the culture container 2 and the lid 3 to ensure airtightness between the culture container 2 and the lid 3. The rubber sheet 5 has a rectangular shape in plan view, with the X-axis direction as the short side and the Y-axis direction as the long side. The upper surface 50A and lower surface 50B of the rubber sheet 5 are both smooth surfaces. The upper surface 50A of the rubber sheet 5 adheres tightly to the smooth surface 30B of the lid 3. In other words, the rubber sheet 5 adheres tightly to the lid 3 side due to van der Waals forces.

[0065] The rubber sheet 5 is provided with connecting holes 51a and 51b that connect grooves 31a and 31b provided in the lid 3 to storage sections 21a and 21b provided in the culture container 2. The rubber sheet 5 is also provided with a second connecting hole 53 between the connecting holes 51a and 51b that communicates with the hydrogel storage tank 116.

[0066] As shown in Figures 7 and 8, the culture vessel 2 is provided with linear sealing protrusions 60 that surround the openings of the multiple storage sections 21 and project from the upper surface 20A toward the rubber sheet 5. The sealing protrusions 60 are provided along the respective opening edges of the storage sections 21a, 21b and the hydrogel storage tank 116. At least a portion of the adjacent sealing protrusions 60 of the storage sections 21a, 21b and the hydrogel storage tank 116 are common to each other.

[0067] In the cell culture apparatus 1 with the above configuration, the culture vessel 2, which has storage sections 21a and 21b, is equipped with a lid 3 that covers the culture vessel 2 in order to simultaneously ensure airtightness of the storage sections 21a and 21b. A rubber sheet 5 is placed between the culture vessel 2 and the lid 3 to ensure airtightness of the multiple storage sections 21. At this time, by providing linear sealing protrusions 60 on the upper surface 20A of the culture vessel 2, the rubber sheet 5 is elastically deformed, and airtightness can be ensured in the area along the sealing protrusions 60. With this rubber sheet 5, airtightness of multiple storage sections 21 can be ensured with a single component, making assembly easy, and by adjusting the thickness T of the rubber sheet 5 and the height H of the sealing protrusions 60, dimensional errors of the culture vessel 2 and the lid 3 can be absorbed and airtightness can be easily ensured.

[0068] Furthermore, according to the second embodiment of the above configuration, culture medium 102 is stored in storage sections 21a and 21b adjacent to the hydrogel chamber 110, and by applying pneumatic pressure to the stored culture medium 102, the culture medium 102 can permeate the hydrogel 101 and cells can be cultured. In this way, by fixing a lid 3 connected to a pneumatic pipe 4 to the culture vessel 2 and culturing cells (for example, vascular endothelial cells) while permeating the hydrogel 101 with culture medium 102 using pneumatic pressure, the formation of cell tissue in the hydrogel 101 can be promoted.

[0069] While preferred embodiments and examples of the present invention have been described and explained above, it should be understood that these are illustrative examples of the present invention and should not be considered as limiting. Additions, omissions, substitutions, and other modifications can be made without departing from the scope of the invention. Therefore, the present invention should not be considered limited by the foregoing description, but rather limited by the claims. [Explanation of Symbols]

[0070] 1...Cell culture device 2…Culture container 2a… Upper plate 2b...Bottom plate 2c...Flow plate 2D…Transparent plate 3…Lid 3a…Lid body 3b...cover 3c... Rubber sheet 4…Pneumatic piping 4a~4d...Pneumatic piping 5…Rubber sheet 6…First fixing part 7...Second fixing part 8...First locking part 9…Gap 10...Second locking part 11... Bolt 12…Through hole 13…Window section 14…Through hole 15… Discharge channel 16…Inlet channel 20A…Top surface 21... Storage section 21a~21d...Storage section 22…Positioning recess 30a…Connection hole 30A…Top surface 30B…Smooth surface 31… Groove 31a~31d...Groove 32…Through hole 33…Positioning protrusion 50A…Top surface 50B…Bottom surface 51...Communication hole 51a~51d...Communication hole 53...Second communication hole 60... Seal protrusion 61... Short side 62... Long side 63... Corner 64…Space 70…Opening 101... Hydrogel 102...Culture solution 110... Hydrogel Chamber 114…Inlet hole 116... Hydrogel storage tank 122…Strut row 132…Strut row 151... Laplace valve

Claims

1. A culture vessel having multiple storage compartments with openings, A lid that is detachably attached to the culture vessel and covers the openings of the plurality of storage sections, A pneumatic pipe connected to either the culture vessel or the lid, A rubber sheet is placed between the culture vessel and the lid to ensure airtightness between the culture vessel and the lid, The facility comprises linear sealing protrusions that surround the openings of the plurality of storage sections and project toward the rubber sheet, Either the culture container or the lid, and the sealing projection are formed from a single component. Cell culture equipment.

2. The sealing projection is provided on the culture vessel. The cell culture apparatus according to claim 1.

3. The lid has a smooth surface on which the rubber sheet adheres. The cell culture apparatus according to claim 2.

4. The lid has a plurality of positioning protrusions for positioning the rubber sheet on the smooth surface. The cell culture apparatus according to claim 3.

5. The plurality of positioning protrusions are provided on both sides of the rubber sheet in the short direction and on one side of the rubber sheet in the longitudinal direction. The cell culture apparatus according to claim 4.

6. The cell culture apparatus according to claim 4 or 5, wherein the culture vessel comprises a plurality of positioning recesses into which the plurality of positioning protrusions are inserted.

7. The aforementioned pneumatic piping is connected to the cover, A cell culture apparatus according to any one of claims 1 to 5.

8. The cover is provided with a plurality of grooves that communicate with the pneumatic piping, The rubber sheet is provided with a plurality of communication holes that connect the plurality of grooves and the plurality of storage portions. The cell culture apparatus according to claim 7.

9. The sealing projection is provided along the opening edge of each of the plurality of storage sections. A cell culture apparatus according to any one of claims 1 to 5.

10. Of the plurality of storage sections, at least a portion of the sealing protrusions provided along the opening edges of adjacent storage sections are common to each other. The cell culture apparatus according to claim 9.

11. The height / width ratio of the sealing protrusion is the same in the common portion and the non-common portion. The cell culture apparatus according to claim 10.

12. The ratio of the height / width of the aforementioned sealing protrusion is within the range of 0.5 to 10. A cell culture apparatus according to any one of claims 1 to 5.

13. The aforementioned rubber sheet is made of silicone rubber. A cell culture apparatus according to any one of claims 1 to 5.

14. The durometer hardness of the aforementioned rubber sheet is 50 or less. A cell culture apparatus according to any one of claims 1 to 5.

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