Cell culture container

Abstract

Provided is a cell culture vessel suited to improving difficulties when performing perfusion culture in a closed state. The cell culture vessel is equipped with a vessel body 1 having an opening 110 on the upper end and a first cylindrical portion 11 extending vertically, and a plug member 2 capable of sealing the opening 110. The plug member 2 includes: a second cylindrical portion 21 that abuts the inner peripheral surface and closes the first cylindrical portion 11; a first flange portion 22 that extends radially outward from the upper end of the second cylindrical portion 21 and blocks the upper end 114 of the first cylindrical portion 11; and a first bottom portion 23 that extends radially inward from the lower end of the second cylindrical portion 21 and blocks the inside of the second cylindrical portion 21 when viewed in the vertical direction. The first bottom portion 23 is provided with a plurality of ports 24 each having a flow path 242 leading to the outside and the inside space of the vessel body 1. The first bottom portion 23 has a ceiling surface 231 that faces downward, and ceiling recesses 232 recessed upward from the ceiling surface 231 are formed in the first bottom portion 23. The ceiling recesses 232 connect to the flow paths 242.

Description

[Technical Field]

[0001] The present disclosure relates to a cell culture vessel for culturing cells, and in particular to a cell culture vessel used for perfusion culture that automatically and constantly supplies a culture medium at a constant, low rate and simultaneously discharges an equal amount of the culture medium. [Background technology]

[0002] Culture vessels such as dishes, well plates, and flasks are widely used for cell culture. Even with lids, dishes and well plates have gaps when placed on them, making them impossible to seal. While flasks can be sealed, all of these conventional culture vessels require the lid to be opened and closed for tasks such as changing the culture medium. Therefore, while these culture vessels are highly versatile for cell culture applications for research purposes, they pose problems for medical cell culture applications, including the risk of contamination, the need for specialized personnel and costs, and the difficulty of standardizing the quality of cultured cells.

[0003] Recently, various automatic culture devices have been developed to solve these problems (see, for example, Patent Document 1). A culture vessel used in an automatic culture device is provided with, for example, a pair of ports. These ports are provided, for example, on the lid of the culture vessel, and each port extends downward from the lid and is positioned inside the culture vessel. When culturing cells, new culture medium is supplied to the culture vessel through one port, and the culture medium inside the culture vessel is discharged to the outside through the other port. To supply culture medium from one port and discharge it from the other port, simply connect appropriate pumps to each port. In this case, the liquid will flow even if there is a layer of culture medium and gas inside the vessel. However, if the amount of culture medium supplied from one port and the amount of culture medium discharged from the other port do not exactly match, the liquid in the vessel will run out or overflow if the culture is continued for a long period of time.

[0004] To avoid these problems, one effective method is to use a single pump per vessel, supplying culture medium through one port under positive pressure and allowing it to overflow and drain through the other port, or draining culture medium through one port under negative pressure and drawing it in through the other port. Doing this all at once would cause turbulence and damage the cells, so perfusion culture is performed by automatically supplying culture medium at a constant, slow rate and simultaneously draining an equal amount of culture medium. To achieve smooth fluid flow in perfusion culture, it is important to maintain a closed system and to fill the entire interior of the culture vessel with culture medium, avoiding the formation of gas layers or bubbles. However, even when attempting to fill a closed-system culture vessel with culture medium, small amounts of air (air bubbles) may remain inside the vessel. When the port extends downward from the lid and is positioned inside the vessel, as described above, it is not possible to evacuate the air remaining inside the vessel. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2017-79633 Summary of the Invention [Problem to be solved by the invention]

[0006] The present disclosure was conceived under these circumstances, and its main objective is to provide a cell culture vessel suitable for overcoming the inconveniences associated with perfusion culture in a closed system. [Means for solving the problem]

[0007] In order to solve the above problems, the present disclosure employs the following technical means.

[0008] A cell culture vessel provided by the present disclosure includes a vessel body having an opening at one end in a first direction and a first cylindrical portion extending in the first direction, and a plug member capable of closing the opening, wherein the plug member includes a second cylindrical portion abutting against an inner circumferential surface of the first cylindrical portion to seal the opening, a first flange portion extending radially outward from the one end in the first direction of the second cylindrical portion and closing the one end in the first direction of the first cylindrical portion, and a second flange portion extending radially inward from the other end in the first direction of the second cylindrical portion and closing the one end in the first direction of the first cylindrical portion, and a plate-shaped first bottom portion that closes the inside of the second cylindrical portion, the first bottom portion having a plurality of ports each having a flow path that is open to the outside at one side end in the first direction and is open to the internal space of the container body at the other side end in the first direction, the first bottom portion having a ceiling surface facing the other side in the first direction, and a ceiling recess that is recessed from the ceiling surface to the one side in the first direction, and the ceiling recess connected to the flow path in at least any one of the plurality of ports.

[0009] In a preferred embodiment, the ceiling recess communicates with the outer periphery of the first bottom portion.

[0010] In a preferred embodiment, the container further comprises an outer lid having an annular top plate portion and a third cylindrical portion extending from the outer peripheral edge of the top plate portion in the thickness direction of the top plate portion and fitted onto the first cylindrical portion, and at least one of the container body and the outer lid is provided with a locking means for preventing relative movement between the first cylindrical portion and the third cylindrical portion when the first flange portion abuts against the first cylindrical portion.

[0011] In a preferred embodiment, the plug member is made of a soft material.

[0012] In a preferred embodiment, the device further comprises a cover member made of a hard material and overlapping and abutting the plug member.

[0013] In a preferred embodiment, the cover member includes a second flange portion that covers the first flange portion, a fourth cylindrical portion that covers the inner surface of the second cylindrical portion, and a second bottom portion that covers at least a portion of the first bottom portion.

[0014] In a preferred embodiment, the port has an extension portion extending from the first bottom portion to one side in the first direction, and the cover member includes a fifth cylindrical portion abutting against and surrounding an outer peripheral surface of the extension portion.

[0015] In a preferred embodiment, the first bottom portion includes a thin portion that is relatively thinner than other portions and has gas permeability.

[0016] In a preferred embodiment, the first bottom portion includes a thin-walled portion that is relatively thinner than other portions and has gas permeability, and the second bottom portion has a through hole formed therein that penetrates in the first direction, and the through hole overlaps with the thin-walled portion when viewed in the first direction.

[0017] In a preferred embodiment, the tip of the extension is provided with a protrusion that protrudes outward from the outer circumferential surface and is capable of abutting against the one side end of the fifth cylindrical portion in the first direction.

[0018] In a preferred embodiment, the flow path comprises a first flow path that is open to the outside and a second flow path that is open to the internal space of the container body and is connected to the ceiling recess, and the first flow path and the second flow path are spaced apart from each other in the first direction and overlap when viewed in the first direction.

[0019] Other features and advantages of the cell culture vessel according to the present disclosure will become more apparent from the following detailed description taken in conjunction with the accompanying drawings. [Brief explanation of the drawings]

[0020] [Figure 1] FIG. 1 is a perspective view showing a cell culture vessel according to a first embodiment. [Figure 2] FIG. 2 is an exploded perspective view of the cell culture vessel shown in FIG. [Figure 3] FIG. 2 is a plan view of the cell culture vessel shown in FIG. [Figure 4] FIG. 4 is an enlarged cross-sectional view taken along line IV-IV in FIG. 3. [Figure 5] FIG. 4 is an enlarged cross-sectional view taken along line VV in FIG. 3. [Figure 6] FIG. 2 is a bottom view of the plug member that constitutes the cell culture vessel according to the first embodiment. [Figure 7] FIG. 5 is a cross-sectional view similar to FIG. 4, showing the cell culture vessel according to the first embodiment in use. [Figure 8] FIG. 5 is a cross-sectional view similar to FIG. 4, showing the cell culture vessel according to the first embodiment in use. [Figure 9] FIG. 5 is a cross-sectional view similar to FIG. 4, showing the cell culture vessel according to the first embodiment in use. [Figure 10] 5 is a cross-sectional view similar to FIG. 4, showing a cell culture vessel according to a first modified example of the first embodiment. [Figure 11] FIG. 10 is a bottom view of a plug member constituting a cell culture vessel according to a first modified example of the first embodiment. [Figure 12] FIG. 10 is a bottom view showing another example of the plug member. [Figure 13] 5 is a cross-sectional view similar to FIG. 4, showing a cell culture vessel according to a second modified example of the first embodiment. FIG. [Figure 14] 5 is a cross-sectional view similar to FIG. 4, showing a cell culture vessel according to a third modified example of the first embodiment. FIG. [Figure 15] 5 is a cross-sectional view similar to FIG. 4, showing a cell culture vessel according to a fourth modified example of the first embodiment. [Figure 16] FIG. 5 is a cross-sectional view similar to FIG. 4, showing a cell culture vessel according to a second embodiment. [Figure 17] FIG. 5 is a cross-sectional view similar to FIG. 4, showing a cell culture vessel according to a third embodiment. [Figure 18] FIG. 5 is a cross-sectional view similar to FIG. 4, showing a cell culture vessel according to a fourth embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0021] Preferred embodiments of the present disclosure will now be described in detail with reference to the drawings.

[0022] In this disclosure, unless otherwise specified, "an object A overlaps an object B when viewed in a certain direction" includes "an object A overlaps the entire object B" and "an object A overlaps a part of an object B."

[0023] Terms such as "first," "second," and "third" in this disclosure are used merely as labels and are not necessarily intended to dictate any ordering of their objects.

[0024] First Embodiment 1 to 5 show a cell culture vessel according to a first embodiment of the present disclosure. The cell culture vessel A1 of this embodiment includes a vessel body 1, a plug member 2, a cover member 3, and an outer lid 4. As will be described in detail later, FIGS. 1 and 3 to 5 show an assembled state in which the vessel body 1, plug member 2, cover member 3, and outer lid 4 are combined. FIG. 2 is an exploded perspective view of the components of the cell culture vessel A1.

[0025] In this embodiment, the container body 1 includes a first cylindrical portion 11 and a container bottom portion 12, and is container-shaped with an opening 110 at an upper end 114 (one end in the first direction) of the first cylindrical portion 11. The first cylindrical portion 11 is roughly cylindrical and extends in the up-down direction (first direction). A male thread 111 is formed on the outer peripheral surface of the upper portion of the first cylindrical portion 11. The container bottom portion 12 is connected to the vicinity of the lower end of the first cylindrical portion 11 (near the other end in the first direction) and closes the inside of the first cylindrical portion 11. In this embodiment, a slit 112 is formed at an appropriate position at the lower end of the first cylindrical portion 11. Also, in this embodiment, an extension piece 113 is provided at an appropriate position at the lower end of the first cylindrical portion 11. The extension piece 113 is a portion that is connected to the first cylindrical portion 11 and extends radially outward. The extension piece 113 is provided at a position different from the slit 112 in the circumferential direction of the first cylindrical portion 11.

[0026] The container body 1 is formed of, for example, a translucent or transparent plastic material, such as polystyrene, methylpentene, polycarbonate, cycloolefin polymer, cycloolefin copolymer, or other preferably transparent materials, but is not limited to these.

[0027] The upper surface of the container bottom 12 is a flat cell culture surface for culturing cells. This cell culture surface (the upper surface of the container bottom 12) may be subjected to a surface treatment to improve cell adhesiveness, as needed. Examples of such surface treatments include hydrophilization treatments such as corona discharge treatment and plasma treatment.

[0028] The plug member 2 closes the opening 110 of the container 1 to seal the container 1. The plug member 2 is, for example, a rubber molded product, and includes a second tubular portion 21, a first flange portion 22, a first bottom portion 23, and multiple ports 24, as shown in FIGS. 2, 4, 5, etc. The second tubular portion 21 is generally cylindrical and abuts against the inner circumferential surface of the first tubular portion 11 to seal the container 1. In this embodiment, a pair of annular protrusions 211 are formed on the outer circumferential portion of the second tubular portion 21, spaced apart in the vertical direction (first direction). For example, the outer diameter of the annular protrusions 211 in its natural state is slightly larger than the inner diameter of the first tubular portion 11. When the plug member 2 is attached to the container 1, the second tubular portion 21 is fitted into the first tubular portion 11 while being compressed radially inward. The annular protrusions 211 then tightly contact the inner circumferential surface of the first tubular portion 11 to seal the container 1. The second cylindrical portion 21 may not have the annular protrusion 211. When the second cylindrical portion 21 does not have the annular protrusion 211, the outer peripheral surface of the second cylindrical portion 21 comes into surface contact with the inner peripheral surface of the first cylindrical portion 11 to form a tight seal.

[0029] The first flange portion 22 is generally annular, and extends radially outward from the upper end (one side end in the first direction) of the second cylindrical portion 21. The first flange portion 22 overlaps with the first cylindrical portion 11 when viewed in the thickness direction of the first flange portion 22, and closes the upper end of the first cylindrical portion 11. The first flange portion 22 has an appropriate thickness and has an appropriate elastic restoring force against a load in the vertical direction (first direction). The thickness of the first flange portion 22 is, for example, about 1 to 3 mm.

[0030] The first bottom portion 23 extends radially inward from the lower end (the other end in the first direction) of the second cylindrical portion 21 and closes the inside of the second cylindrical portion 21 when viewed in the up-down direction. The first bottom portion 23 is plate-shaped and has a ceiling surface 231. In this embodiment, the ceiling surface 231 is a flat surface facing downward (the other side in the first direction). A ceiling recess 232 is formed in the first bottom portion 23. The ceiling recess 232 is a portion recessed upward (to one side in the first direction) from the ceiling surface 231. The ceiling recess 232 is provided near the outer periphery of the first bottom portion 23. As will be described in detail later, the ceiling recess 232 is arranged to correspond to the port 24.

[0031] In this embodiment, the first bottom portion 23 has a thin-walled portion 238. As shown in FIGS. 4, 5, etc., the thin-walled portion 238 is a portion that is thinner than other portions. The thin-walled portion 238 is disposed in the center of the first bottom portion 23 and has a substantially circular shape when viewed in the up-down direction (first direction). In this embodiment, the top surface of the first bottom portion 23 is recessed in the center, and the recessed portion of the top surface of the first bottom portion 23 is the thin-walled portion 238. In FIG. 6, the boundary between the thin-walled portion 238 and a portion surrounding the thin-walled portion 238 when viewed in the thickness direction (first direction) (hereinafter referred to as the "outer peripheral thick-walled portion 235") is represented by a virtual line.

[0032] In this embodiment, the thin portion 238 is gas permeable. Also in this embodiment, the thin portion 238 is transparent. The thickness of the thin portion 238 is, for example, about 0.2 to 0.3 mm. The thickness of the outer peripheral thick portion 235 surrounding the thin portion 238 is approximately the same as the thickness of the first flange portion 22, and is, for example, about 1 to 3 mm.

[0033] The multiple ports 24 are provided in the first bottom portion 23 and are integrally formed at appropriate positions on the first bottom portion 23. In the present embodiment, a pair of ports 24 is provided in the first bottom portion 23. The pair of ports 24 are provided near the outer periphery of the first bottom portion 23 and are arranged on opposite sides of the center of the first bottom portion 23 when viewed in the up-down direction (first direction). Each port 24 is provided in the outer periphery thick portion 235.

[0034] The port 24 is used to supply the culture medium into the container 1 or to discharge the culture medium from the container 1 to the outside, and has an extension 241 and a flow path 242 as shown in Fig. 4. The extension 241 is cylindrical and extends upward (to one side in the first direction) from the first bottom 23, and has an outer circumferential surface 241a with a circular cross section. In this embodiment, the tip of the extension 241 is provided with a protrusion 245 that protrudes outward beyond the outer circumferential surface 241a.

[0035] The flow path 242 has an upper end (one side end in the first direction) that is open to the outside and a lower end (hand side end in the first direction) that is open in the internal space of the container body 1, and is a communication path that leads between the outside and the internal space of the container body 1. In this embodiment, the flow path 242 has a circular cross section and is formed in the extension portion 241.

[0036] The ceiling recess 232 overlaps with the port 24 when viewed in the up-down direction (first direction) and is connected to the flow path 242. In this embodiment, the ceiling recess 232 is conical in shape and is formed so that the area of ​​its cross section increases downward from the position where it connects to the flow path 242. In the example illustrated in this embodiment, the ceiling recess 232 is formed spanning from the first bottom portion 23 to the port 24 (extension portion 241).

[0037] The plug member 2 having the above configuration is made of a soft material that is flexible and elastic. Examples of materials that can be used to make the plug member 2 include silicone rubber and elastomer resin. As will be described in detail later, considering the contact between the plug member 2 and the contents (e.g., culture medium), medical-grade silicone rubber, which is non-cytotoxic and biocompatible, is more preferable as the material for the plug member 2. Furthermore, the hardness of the plug member 2 is preferably, for example, a rubber hardness of about 20 to 40 degrees.

[0038] The cover member 3 overlaps the plug member 2 and abuts against the plug member 2. In this embodiment, the cover member 3 includes a second flange portion 31, a fourth cylindrical portion 32, a second bottom portion 33, and a fifth cylindrical portion 34.

[0039] 4 and 5 , the second flange portion 31 covers the first flange portion 22 from above and abuts against the upper surface of the first flange portion 22. The fourth tubular portion 32 is connected to the inner peripheral edge of the second flange portion 31. The fourth tubular portion 32 is roughly cylindrical and extends downward (toward the other side in the first direction) from the inner peripheral edge of the second flange portion 31. The fourth tubular portion 32 covers the inner peripheral surface of the second tubular portion 21 and abuts against the inner peripheral surface of the second tubular portion 21.

[0040] The second bottom portion 33 is connected to the lower end (the other end in the first direction) of the fourth cylindrical portion 32 and extends radially inward from the lower end of the fourth cylindrical portion 32. The second bottom portion 33 covers the first bottom portion 23 from above and abuts against the upper surface of the first bottom portion 23. In this embodiment, the central portion of the second bottom portion 33 is recessed downward (to the other side in the first direction) in correspondence with the location where the thin-walled portion 238 is formed.

[0041] A through hole 331 is formed in the second bottom portion 33. The through hole 331 penetrates the second bottom portion 33 in the thickness direction and overlaps with the thin-walled portion 238 of the plug member 2 when viewed in the up-down direction (first direction). In this embodiment, a plurality of through holes 331 are formed in the second bottom portion 33. As shown in FIG. 3 and other figures, the plurality of through holes 331 are arranged so as to be appropriately dispersed when viewed in the up-down direction.

[0042] The fifth cylindrical portion 34 is provided on the second bottom portion 33 and extends upward (to one side in the first direction) from an appropriate position on the second bottom portion 33. In the present embodiment, a pair of fifth cylindrical portions 34 is provided on the second bottom portion 33. The pair of fifth cylindrical portions 34 is arranged to correspond to each of the pair of ports 24. In the present embodiment, each fifth cylindrical portion 34 is shaped to connect to a part of the fourth cylindrical portion 32. A port 24 (extension portion 241) is fitted into each of the pair of fifth cylindrical portions 34. Specifically, the inner diameter dimension of the fifth cylindrical portion 34 is slightly smaller than the outer diameter dimension of the extension portion 241 in a natural state. Each extension portion 241 is press-fitted into the fifth cylindrical portion 34. As a result, the fifth cylindrical portion 34 is in close contact with (abuts against) the outer peripheral surface 241a of the extension portion 241 and surrounds the outer peripheral surface 241a.

[0043] 4 and other figures, in this embodiment, when viewed in the up-down direction (first direction), the fifth cylindrical portion 34 overlaps with a protrusion 245 provided at the tip of the port 24. The protrusion 245 is adjacent to and above the fifth cylindrical portion 34. This allows the protrusion 245 to abut against the upper end of the fifth cylindrical portion 34.

[0044] The cover member 3 is made of a hard material, such as a translucent or transparent plastic material, such as polystyrene, methylpentene, polycarbonate, cycloolefin polymer, or cycloolefin copolymer, preferably having transparency, but not limited to these.

[0045] The outer lid 4 has a top plate portion 41 and a third cylindrical portion 42. The top plate portion 41 is generally annular and is a portion that sandwiches the second flange portion 31 of the cover member 3 between it and the upper end 114 of the first cylindrical portion 11 of the container body 1. A through hole 411 is formed on the radially inner side of the top plate portion 41. When viewed in the thickness direction of the top plate portion 41, the through hole 411 is surrounded by the top plate portion 41. When viewed in the thickness direction (up and down direction) of the top plate portion 41, the through hole 411 overlaps with the thin-walled portion 238 of the plug member 2 and the multiple through holes 331 in the cover member 3.

[0046] The third cylindrical portion 42 is generally cylindrical and extends from the outer peripheral edge of the top plate portion 41 in the thickness direction (downward in the figure) of the top plate portion 41. A female thread 421 is formed on the inner peripheral surface of the third cylindrical portion 42, and the female thread 421 can be threaded onto the male thread 111 of the container body 1 (first cylindrical portion 11).

[0047] In this embodiment, the top plate 41 is provided with a convex portion 412. As clearly shown in FIG. 5, the convex portion 412 protrudes upward (to one side in the first direction) from the main surface 41a of the top plate 41, which faces the thickness direction. In this embodiment, a plurality of convex portions 412 are provided and separated from one another. A plane P1 formed by the tips 412a of these convex portions 412 is substantially parallel to the main surface 41a of the top plate 41. Here, "substantially parallel" is intended to mean that the plane P1 and the main surface 41a are designed to be parallel, but also includes cases where there is some variation in parallelism due to dimensional tolerances, etc. In addition, the plane P1 formed by the tips 412a of the plurality of convex portions 412 is located at the uppermost position in the cell culture vessel A1. As can be seen from FIGS. 1 to 5, in this embodiment, the convex portions 412 are arranged in the shape of concentric arcs when viewed in the vertical direction, and there is a gap between adjacent convex portions 412. As a result, the convex portion 412 does not form a closed ring when viewed in the vertical direction (thickness direction of the top plate portion 41).

[0048] 4 and 5, a skirt portion 121 that protrudes downward from the outer periphery of the vessel bottom 12 of the vessel body 1 is connected to the vessel bottom 12. The outer diameter of the outer periphery of the concentric arc formed by the plurality of convex portions 412 is slightly smaller than the inner diameter of the skirt portion 121. With this configuration, the convex portion 412 of one cell culture vessel A1 can be fitted into the skirt portion 121 of another cell culture vessel A1, allowing the other cell culture vessels A1 to be stacked. Therefore, multiple cell culture vessels A1 can be stacked in a stable position.

[0049] The outer cover 4 is made of, for example, an opaque, translucent, or transparent plastic material, such as polyethylene, polypropylene, polystyrene, methylpentene, or polycarbonate, but is not limited to these.

[0050] Next, the method of use and the action of the cell culture vessel A1 will be described with reference to FIGS.

[0051] The cell culture vessel A1 is used for perfusion culture, which automatically supplies a culture medium at a constant, low rate and simultaneously discharges an equal amount of the culture medium. Note that the cultured cells and culture medium contained in the cell culture vessel A1 (vessel body 1) are not particularly limited.

[0052] In cell culture using the cell culture vessel A1, the plug member 2, cover member 3, each port 24, and connector 7 are assembled in advance (see Figure 7). The connectors 7 are press-fitted into each of the pair of ports 24. A perfusion pump (not shown) is connected to one of the ports 24 via the connector 7 and tubing 8, and a drainage container (not shown) is connected to the other port 24 via the connector 7 and tubing 8. Each tube 8 is passed through the inside of the outer lid 4 in advance. Next, cells to be cultured are seeded in the vessel body 1, and a predetermined amount of culture medium is poured into the vessel body 1. Next, the vessel body 1 is covered with the plug member 2 and cover member 3, and the outer lid 4 is fastened.

[0053] Next, the culture medium is supplied to the cell culture vessel A1 and air is discharged through a pair of ports 24 and the connector 7 connected to them, filling the vessel body 1 with the culture medium. Here, the culture medium is supplied into the vessel body 1 through one port 24, while the air inside the vessel body 1 is discharged to the outside through the other port 24. FIG. 8 shows the state in which the vessel body 1 is filled with the culture medium C. The culture medium C fills the inner space of the vessel body 1 (the space surrounded by the vessel bottom 12 and first cylindrical portion 11 of the vessel body 1 and the first bottom 23 of the plug member 2). At this time, a small amount of air (hereinafter referred to as "air bubbles B") may remain in the inner space of the vessel body 1.

[0054] In the cell culture vessel A1, the ceiling surface 231 of the first bottom 23 of the plug member 2 is flat. A ceiling recess 232 recessed upward from the ceiling surface 231 is formed in the first bottom 23, and the ceiling recess 232 is connected to the flow path 242 of the port 24. With this configuration, when the cell culture vessel A1 is tilted so that the port 24 is positioned at the top, as shown in FIG. 9 , bubbles B in the vessel 1 move along the ceiling surface 231 to the vicinity of the port 24. By operating the perfusion pump in this state, bubbles B remaining in the vessel 1 further move through the ceiling recess 232 into the flow path 242 and can be discharged to the outside of the cell culture vessel A1. When perfusion culture is performed, fresh culture solution C is supplied to the vessel 1 (cell culture vessel A1) through one port 24, and the culture solution C in the vessel 1 (inside the cell culture vessel A1) is discharged to the outside through the other port 24. In the perfusion culture step, the culture solution C in the vessel body 1 is continuously replaced little by little.

[0055] The cell culture vessel A1 includes an outer lid 4 in addition to a vessel body 1 and a stopper member 2. A female thread 421 is formed in the third cylindrical portion 42 of the outer lid 4, which is fitted onto the first cylindrical portion 11 of the vessel body 1. By tightening the female thread 421 into the male thread 111 of the vessel body 1 (first cylindrical portion 11), the first flange portion 22 of the stopper member 2 is sandwiched between the upper end 114 of the first cylindrical portion 11 and the top plate portion 41 of the outer lid 4, maintaining the assembled state. The stopper member 2 is a continuous membrane extending from the first flange portion 22 on the outer periphery to the first bottom portion 23 and the thin-walled portion 238 in the center. This liquid-seals the contents (cultured cells and culture solution C) contained in the vessel body 1. Therefore, in the perfusion culture process, the culture solution perfusion operation can be performed while maintaining the cell culture vessel A1 in a closed state, thereby avoiding the risk of contamination. Therefore, the cell culture vessel A1 can maintain the quality of the cells by maintaining a good state of the culture solution C. The male thread 111 of the vessel body 1 (first cylindrical portion 11) and the female thread 221 of the outer lid 4 (third cylindrical portion 42) correspond to an example of the "locking means" of the present disclosure.

[0056] The cell culture vessel A1 includes a vessel body 1, a plug member 2, and a cover member 3. The plug member 2 is made of a soft material. The cover member 3 is made of a hard material and overlaps and abuts against the plug member 2. Specifically, the cover member 3 includes a second flange portion 31 that covers the first flange portion 22 of the plug member 2. With this configuration, the second flange portion 31 is interposed between the first flange portion 22 of the plug member 2 and the top plate portion 41 of the outer lid 4. Therefore, when the female thread 421 of the outer lid 4 is tightened onto the male thread 111 of the vessel body 1 (first cylindrical portion 11), the top plate portion 41 comes into sliding contact with the second flange portion 31 (hard material), improving the slidability of the outer lid 4.

[0057] The cover member 3 includes a fourth cylindrical portion 32 that covers the inner circumferential surface of the second cylindrical portion 21. With this configuration, the plug member 2 made of a soft material is pressed against the inner circumferential surface of the container body 1 (first cylindrical portion 11), improving the sealing of the cell culture container A1. The cover member 3 also includes a second bottom portion 33 that covers the first bottom portion 23 of the plug member 2. With this configuration, during the perfusion culture step, the first bottom portion 23 (thin-walled portion 238) tends to swell upward due to the pressure of the supplied culture solution C, but the second bottom portion 33 suppresses deformation of the first bottom portion 23 (thin-walled portion 238). This allows the culture solution C to flow smoothly within the container body 1 and provides excellent visibility inside the container body 1, making it suitable for observing the conditions within the container body 1.

[0058] The port 24 provided in the plug member 2 has an extending portion 241 extending upward from the first bottom portion 23. The cover member 3 also includes a fifth cylindrical portion 34, which abuts against an outer peripheral surface 241a of the extending portion 241 and covers the outer peripheral surface 241a. With this configuration, when the connector 7 is press-fitted into the port 24 (extending portion 241), the fifth cylindrical portion 34 prevents the extending portion 241 from expanding radially outward. As a result, the sealing performance when the connector 7 is connected to the port 24 is improved.

[0059] The first bottom portion 23 of the plug member 2 includes a gas-permeable thin-walled portion 238. This maintains the content of the container body 1 (cell culture vessel A1) in a state of ventilation with the outside of the container body 1 (cell culture vessel A1). Therefore, according to this embodiment, the content of the container body 1 (cell culture vessel A1) can be cultured in a state of ventilation with the outside. Furthermore, by placing the cell culture vessel A1 in an incubator under a predetermined gas atmosphere, the gas atmosphere in the incubator can be taken into the container body 1 (cell culture vessel A1).

[0060] A through-hole 331 is formed in the second bottom 33 of the cover member 3, penetrating the second bottom 33 in the thickness direction. The through-hole 331 overlaps with the thin-walled portion 238 of the plug member 2 when viewed in the up-down direction (first direction). With this configuration, it is possible to maintain ventilation between the inside and outside of the container body 1 via the thin-walled portion 238, while suppressing deformation of the first bottom 23 (thin-walled portion 238) as described above.

[0061] A protrusion 245 that protrudes outward from the outer peripheral surface 241a of the extension 241 is provided at the tip of the port 24 (extension 241). The protrusion 245 is adjacent to the upper part of the fifth cylindrical portion 34 and can abut against the upper end of the fifth cylindrical portion 34. With this configuration, even if a downward pressing force acts on the extension 241, the extension 241 is prevented from being pushed into the internal space of the container 1 through the fifth cylindrical portion 34. As a result, for example, when the connector 7 is press-fitted into the port 24, a downward pressing force acts on the port 24, but the extension 241 is prevented from intruding into the internal space of the container 1, and damage to cells in the container 1 is avoided.

[0062] <First Modification of First Embodiment> Fig. 10 shows a cell culture vessel according to a first modified example of the first embodiment. In the figures following Fig. 10, elements that are the same as or similar to those in the cell culture vessel A1 of the above embodiment are denoted by the same reference numerals as in the above embodiment, and descriptions thereof will be omitted where appropriate.

[0063] The cell culture vessel A11 of this modified example differs from the cell culture vessel A1 of the above embodiment mainly in the configuration of the ceiling recess 232.

[0064] In this modified example, the ceiling recess 232 has a conical portion 233 and a peripheral portion 234. The conical portion 233 is connected to the flow path 242. In the illustrated example, the conical portion 233 is a cone-shaped portion formed so that the cross-sectional area increases downward from the position where it connects to the flow path 242. The peripheral portion 234 is connected to the conical portion 233 and communicates with an outer peripheral edge 237 of the first bottom 23. The peripheral portion 234 is inclined so that it is positioned downward as it extends radially outward from the first bottom 23. As shown in FIG. 11 , the peripheral portion 234 is roughly fan-shaped and expands radially outward when viewed in the thickness direction (first direction) of the first bottom 23. In addition, in this modified example, the annular protrusion 211 is not formed on the outer peripheral portion of the second cylindrical portion 21, and the outer peripheral surface of the second cylindrical portion 21 abuts against the inner peripheral surface of the first cylindrical portion 11.

[0065] The cell culture vessel A11 of this modified example accommodates cultured cells and a culture solution and is used for perfusion culture, which automatically and constantly supplies the culture solution at a constant, low rate and simultaneously discharges an equal amount of the culture solution. In the cell culture vessel A11, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. The first bottom portion 23 is formed with a ceiling recess 232 that recesses upward from the ceiling surface 231, and the ceiling recess 232 is connected to the flow path 242 of the port 24. With this configuration, even if air bubbles remain in the internal space of the vessel body 1 when the vessel body 1 is filled with culture solution (including cells) prior to perfusion culture, by tilting the cell culture vessel A11 so that the port 24 is positioned at the top, the air bubbles in the vessel body 1 move along the ceiling surface 231 to the vicinity of the port 24. The air bubbles then rise up through the flow path 242 via the ceiling recess 232. This allows air bubbles remaining in the vessel body 1 to be discharged to the outside of the cell culture vessel A11.

[0066] In this modification, the ceiling recess 232 (peripheral edge 234) communicates with the outer periphery 237 of the first bottom 23. With this configuration, when the cell culture vessel A11 is tilted to discharge bubbles remaining in the inner space of the vessel body 1, even if the bubbles reach the outer periphery 237 side of the first bottom 23, the bubbles can be guided along the peripheral edge 234 to the flow path 242. This makes it possible to more accurately discharge bubbles remaining in the inner space of the vessel body 1.

[0067] In addition, the cell culture vessel A11 of this modified example exhibits the same effects as the cell culture vessel A1 described above.

[0068] Fig. 12 is a bottom view showing another example of the plug member 2. The plug member 2 shown in Fig. 12 has a different shape of the peripheral edge portion 234 than the plug member 2 shown in Fig. 11. In the example shown in Fig. 12, the peripheral edge portion 234 is formed to extend along the circumferential direction at the outer periphery of the first bottom portion 23. Like the peripheral edge portion 234 shown in Fig. 11, the peripheral edge portion 234 shown in Fig. 12 is connected to the cone-shaped portion 233 and continues to the outer periphery 237 of the first bottom portion 23. In addition, the peripheral edge portion 234 is inclined so as to be positioned downward as it extends radially outward from the first bottom portion 23.

[0069] 12 is configured with the plug member 2, similar to the cell culture vessel A11 described above, even if bubbles remain in the inner space of the vessel body 1 when the inner space is filled with culture solution (containing cells), the bubbles can be guided along the peripheral edge 234 to the flow path 242. This makes it possible to more accurately discharge bubbles remaining in the inner space of the vessel body 1.

[0070] <Second Modification of First Embodiment> FIG. 13 shows a cell culture vessel according to a second modification of the first embodiment. In a cell culture vessel A12 of this modification, the configurations of the first bottom portion 23 of the plug member 2 and the second bottom portion 33 of the cover member 3 differ from those of the cell culture vessel A1 of the above embodiment. In this modification, the first bottom portion 23 (mainly the thin-walled portion 238) is curved and plate-shaped, and is curved so as to be positioned downward (on the other side in the first direction) as it extends radially inward. The ceiling surface 231 is a curved surface that is curved so as to be positioned downward (on the other side in the first direction) as it extends radially inward. In this modification, the second bottom portion 33 is also curved so as to be positioned downward (on the other side in the first direction) as it extends radially inward. In the cell culture vessel A1 of the above embodiment, the portion of the second bottom portion 33 that overlaps with the thin-walled portion 238 of the plug member 2 when viewed in the up-down direction (first direction) is plate-shaped (see FIG. 4). In this modification, the second bottom portion 33 has a curved portion that overlaps with the thin-walled portion 238 of the plug member 2 when viewed in the up-down direction (first direction). The lower surface of the second bottom portion 33 abuts against the thin-walled portion 238. Note that the thin-walled portion 238 may have a curved shape in its natural state, but is not limited to this. For example, the thin-walled portion 238 may be flat in its natural state and be configured to assume a curved shape when the second bottom portion 33 of the cover member 3 is placed over it.

[0071] The cell culture vessel A12 of this modified example accommodates cultured cells and a culture solution and is used for perfusion culture, which automatically and constantly supplies the culture solution at a constant, low rate and simultaneously discharges an equal amount of the culture solution. In the cell culture vessel A12, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is curved. The first bottom portion 23 is formed with a ceiling recess 232 that recesses upward from the ceiling surface 231, and the ceiling recess 232 is connected to the flow path 242 of the port 24. With this configuration, even if air bubbles remain in the inner space of the vessel body 1 when the inner space is filled with culture solution (including cells) prior to perfusion culture, the air bubbles move along the curved ceiling surface 231 toward the outer periphery. Here, when the cell culture vessel A12 is tilted so that the port 24 is positioned at the top, the air bubbles in the vessel body 1 move along the outer periphery of the ceiling surface 231 to the vicinity of the port 24. The air bubbles then rise up through the flow path 242 via the ceiling recess 232. This allows the air bubbles remaining in the vessel body 1 to be discharged to the outside of the cell culture vessel A12.

[0072] In addition, the cell culture vessel A12 of this modified example exhibits the same effects as the cell culture vessel A1 described above.

[0073] <Third Modification of First Embodiment> FIG. 14 shows a cell culture vessel according to a third modified example of the first embodiment. In a cell culture vessel A13 of this modified example, the configuration of one of a pair of ports 24 is different from that of the cell culture vessel A1 of the above embodiment. In this modified example, the port 24 located on the left side of FIG. 14 is the same as that of the above embodiment, but the configuration of the port 24 located on the right side of the figure is different. The port 24 located on the right side of FIG. 14 has a protrusion 247 that extends downward from the first bottom 23. In the first bottom 23, no ceiling recess 232 is formed around the protrusion 247.

[0074] The cell culture vessel A13 of this modified example accommodates cultured cells and a culture solution and is used for perfusion culture, which automatically and constantly supplies the culture solution at a constant, low rate and simultaneously discharges an equal amount of the culture solution. In the cell culture vessel A13, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. The first bottom portion 23 is formed with a ceiling recess 232 that recesses upward from the ceiling surface 231, and the ceiling recess 232 is connected to a flow path 242 of one port 24 (left side in FIG. 14 ). With this configuration, even if air bubbles remain in the internal space of the vessel body 1 when the vessel body 1 is filled with culture solution (including cells) prior to perfusion culture, by tilting the cell culture vessel A13 so that the one port 24 (left side in FIG. 14 ) is positioned at the top, the air bubbles in the vessel body 1 move along the ceiling surface 231 to the vicinity of the one port 24. The air bubbles then rise up through the ceiling recess 232 and into the flow path 242. This allows air bubbles remaining in the vessel body 1 to be discharged to the outside of the cell culture vessel A13. When perfusion culture is performed, new culture solution C is supplied to the vessel body 1 (cell culture vessel A13) through the port 24 located on the right side of Fig. 14, and the culture solution in the vessel body 1 (inside the cell culture vessel A13) is discharged to the outside through the port 24 located on the left side of Fig. 14. During the perfusion culture process, the culture solution in the vessel body 1 is continuously replaced little by little.

[0075] In addition, the cell culture vessel A13 of this modified example exhibits the same effects as the cell culture vessel A1 described above.

[0076] <Fourth Modification of First Embodiment> 15 shows a cell culture vessel according to a fourth modified example of the first embodiment. In a cell culture vessel A14 of this modified example, the configuration of the second bottom portion 33 of the cover member 3 differs from that of the cell culture vessel A1 of the above embodiment. In this modified example, one through-hole 331 is formed in the second bottom portion 33. In this modified example, the diameter of the through-hole 331 is significantly larger than in the above embodiment. The through-hole 331 overlaps with most of the thin-walled portion 238 of the plug member 2 when viewed in the vertical direction (first direction).

[0077] The cell culture vessel A14 of this modified example accommodates cultured cells and a culture solution and is used for perfusion culture, which automatically and constantly supplies the culture solution at a constant, low rate and simultaneously discharges an equal amount of the culture solution. In the cell culture vessel A14, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. The first bottom portion 23 is formed with a ceiling recess 232 that recesses upward from the ceiling surface 231, and the ceiling recess 232 is connected to the flow path 242 of the port 24. With this configuration, even if air bubbles remain in the internal space of the vessel body 1 when the vessel body 1 is filled with culture solution (including cells) prior to perfusion culture, by tilting the cell culture vessel A14 so that the port 24 is positioned at the top, the air bubbles in the vessel body 1 move along the ceiling surface 231 to the vicinity of the port 24. The air bubbles then rise up through the flow path 242 via the ceiling recess 232. This allows air bubbles remaining in the vessel body 1 to be discharged to the outside of the cell culture vessel A14.

[0078] In addition, the cell culture vessel A14 of this modified example has the same effects as the cell culture vessel A1 described above.

[0079] Second Embodiment 16 shows a cell culture vessel according to a second embodiment of the present disclosure. The cell culture vessel A2 of this embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the port 24. In this embodiment, the port 24 has a first flow path 243, a second flow path 244, and a non-penetrating portion 246 instead of the flow path 242 of the above embodiment.

[0080] As described above, in the first embodiment, the flow path 242 was a communication path connecting the outside with the internal space of the container body 1, but in this embodiment, the first flow path 243 and the second flow path 244 are separated from each other in the vertical direction (first direction). The first flow path 243 is located near the upper part of the port 24 and is open to the outside. The second flow path 244 is located near the lower part of the port 24 and is open to the internal space of the container body 1 and is connected to the ceiling recess 232. The first flow path 243 and the second flow path 244 overlap when viewed in the vertical direction (first direction). The non-penetrating portion 246 is a portion interposed between the first flow path 243 and the second flow path 244.

[0081] The cell culture vessel A2 of this embodiment is used for perfusion culture, which contains cultured cells and culture medium and automatically supplies the culture medium at a constant, low rate while simultaneously discharging an equal amount of culture medium. In this embodiment, the introduction of cells and culture medium into the internal space of the vessel body 1 and the discharge of waste liquid are performed, for example, by attaching the plug member 2, cover member 3, and outer lid 4 to the vessel body 1 to assemble the cell culture vessel A1, and then piercing the non-penetration portion 246 of the port 24 with a syringe needle (not shown). Here, the tip of the syringe needle is positioned in the second flow path 244 directly below the non-penetration portion 246.

[0082] In the cell culture vessel A2, the ceiling surface 231 of the first bottom 23 of the plug member 2 is flat. A ceiling recess 232 recessed upward from the ceiling surface 231 is formed in the first bottom 23, and the ceiling recess 232 is connected to the flow path 242 of the port 24. With this configuration, even if air bubbles remain in the internal space of the vessel body 1 when the internal space is filled with culture solution (containing cells) prior to perfusion culture, by tilting the cell culture vessel A2 so that the port 24 is positioned at the top, the air bubbles in the vessel body 1 move along the ceiling surface 231 to the vicinity of the port 24. The air bubbles then move through the ceiling recess 232 into the second flow path 244 and are discharged via the injection needle. This allows air bubbles remaining in the vessel body 1 to be discharged to the outside of the cell culture vessel A2.

[0083] The port 24 provided in the plug member 2 has an extending portion 241 extending upward from the first bottom portion 23. The cover member 3 also includes a fifth cylindrical portion 34, which abuts against an outer peripheral surface 241a of the extending portion 241 and covers the outer peripheral surface 241a. With this configuration, when an injection needle is inserted into the port 24 (non-penetration portion 246), the fifth cylindrical portion 34 prevents the extending portion 241 from expanding radially outward. After the injection needle is removed from the non-penetration portion 246, the hole made by the injection needle closes. The port 24 (non-penetration portion 246) configured as described above has resealability that closes the hole made by the injection needle.

[0084] In addition, the cell culture vessel A2 of this embodiment has the same effects as the cell culture vessel A1 described above.

[0085] <Third embodiment> 17 shows a cell culture vessel according to a third embodiment of the present disclosure. Unlike the cell culture vessel A1 of the first embodiment, the cell culture vessel A3 of this embodiment does not include a cover member 3. That is, the cell culture vessel A3 includes a vessel body 1, a plug member 2, and an outer lid 4.

[0086] In the cell culture vessel A3, the vessel body 1 and the outer lid 4 have the same configurations as those in the first embodiment. On the other hand, the configuration of the plug member 2 is significantly different from that in the first embodiment.

[0087] In this embodiment, the plug member 2 is configured with a first portion 2A and a second portion 2B. The second portion 2B is made of a flexible, elastic soft material and has a second cylindrical portion 21 and a first flange portion 22. The second portion 2B is interposed between the first portion 2A and the first cylindrical portion 11 of the container body 1. In this embodiment, as in the first embodiment, a pair of annular protrusions 211 are formed on the outer periphery of the second cylindrical portion 21 at an interval in the vertical direction (first direction). When the plug member 2 (first portion 2A and second portion 2B) is attached to the container body 1, the second cylindrical portion 21 is compressed radially inward and fitted into the first cylindrical portion 11. The annular protrusions 211 then tightly fit against the inner circumferential surface of the first cylindrical portion 11 to form a tight seal. Examples of materials that can be used to configure the second portion 2B include silicone rubber and elastomer resin.

[0088] The first portion 2A includes a first bottom portion 23, a port 24, an inner cylindrical portion 26, and an upper flange portion 27. The first bottom portion 23 has a substantially constant thickness. Thus, in this embodiment, unlike the first embodiment, the first bottom portion 23 does not have a thin-walled portion 238. The ceiling recess 232 formed in the first bottom portion 23 has a conical portion 233 and a peripheral portion 234. The conical portion 233 is connected to the flow path 242. In the illustrated example, the conical portion 233 is a cone-shaped portion formed so that the cross-sectional area increases downward from the position where it connects to the flow path 242. The peripheral portion 234 is connected to the conical portion 233 and communicates with the outer periphery of the first bottom portion 23. The peripheral portion 234 is inclined downward as it extends radially outward from the first bottom portion 23. The inner cylindrical portion 26 extends upward from the outer peripheral edge of the first bottom portion 23 and has a generally cylindrical shape. The inner cylindrical portion 26 covers the inner peripheral surface of the second cylindrical portion 21.

[0089] The upper flange portion 27 has a generally annular shape and extends radially outward from the upper end of the inner cylindrical portion 26. The upper flange portion 27 overlaps with the first flange portion 22 and the first cylindrical portion 11 when viewed in the thickness direction of the upper flange portion 27.

[0090] The first part 2A is made of a hard material, such as a translucent or transparent plastic material, preferably a transparent material such as, but not limited to, polystyrene, methylpentene, polycarbonate, cycloolefin polymer, or cycloolefin copolymer.

[0091] The cell culture vessel A3 of this embodiment accommodates cultured cells and a culture solution and is used for perfusion culture, which automatically and constantly supplies the culture solution at a constant, low rate and simultaneously discharges an equal amount of the culture solution. In the cell culture vessel A3, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. The first bottom portion 23 is formed with a ceiling recess 232 that recesses upward from the ceiling surface 231, and the ceiling recess 232 is connected to the flow path 242 of the port 24. With this configuration, even if air bubbles remain in the internal space of the vessel body 1 when the vessel body 1 is filled with culture solution (including cells) prior to perfusion culture, by tilting the cell culture vessel A3 so that the port 24 is positioned at the top, the air bubbles in the vessel body 1 move along the ceiling surface 231 to the vicinity of the port 24. The air bubbles then rise up through the flow path 242 via the ceiling recess 232. This allows air bubbles remaining in the vessel body 1 to be discharged to the outside of the cell culture vessel A11.

[0092] In this embodiment, the ceiling recess 232 (peripheral edge 234) communicates with the outer periphery of the first bottom 23. With this configuration, when the cell culture vessel A3 is tilted to discharge bubbles remaining in the internal space of the vessel body 1, even if the bubbles reach the outer periphery of the first bottom 23 or the outer periphery of the second cylindrical portion 21, the bubbles can be guided along the peripheral edge 234 to the flow path 242. This makes it possible to more accurately discharge bubbles remaining in the internal space of the vessel body 1.

[0093] In addition, the cell culture vessel A3 of this embodiment has the same effects as the cell culture vessel A1 of the first embodiment described above with reference to Figures 1 to 9, etc., within the same range of configuration.

[0094] <Fourth embodiment> 18 shows a cell culture vessel according to a fourth embodiment of the present disclosure. Unlike the cell culture vessel A1 of the first embodiment, the cell culture vessel A4 of this embodiment does not include a cover member 3 or an outer lid 4. That is, the cell culture vessel A4 includes a vessel body 1 and a plug member 2.

[0095] In this embodiment, since the outer lid 4 is not provided, the external thread 111 is not formed on the outer peripheral surface of the first cylindrical portion 11 of the container body 1.

[0096] The plug member 2 is, for example, a rubber molded product, and includes a second cylindrical portion 21, a first flange portion 22, a first bottom portion 23, and a pair of ports 24. The plug member 2 is made of a soft material that is flexible and elastic. The material constituting the plug member 2 is the same as that of the first embodiment. In this embodiment, the annular protrusion 211 is not formed on the outer periphery of the second cylindrical portion 21. The outer diameter dimension of the second cylindrical portion 21 is larger than the inner diameter dimension of the first cylindrical portion 11 in its natural state. The second cylindrical portion 21 configured as described above is press-fitted into the first cylindrical portion 11. As a result, the outer periphery of the second cylindrical portion 21 comes into close contact (abuts) with the inner periphery of the first cylindrical portion 11, sealing them. Furthermore, in this embodiment, the first bottom portion 23 does not have a thin-walled portion 238. The ceiling recess 232 formed in the first bottom portion 23 has a cone shape, and is formed across from the first bottom portion 23 to the port 24 (extension portion 241).

[0097] The cell culture vessel A4 of this embodiment accommodates cultured cells and a culture solution and is used for perfusion culture, which automatically and constantly supplies the culture solution at a constant, low rate and simultaneously discharges an equal amount of the culture solution. In the cell culture vessel A4, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. The first bottom portion 23 is formed with a ceiling recess 232 that recesses upward from the ceiling surface 231, and the ceiling recess 232 is connected to the flow path 242 of the port 24. With this configuration, even if air bubbles remain in the internal space of the vessel body 1 when the vessel body 1 is filled with culture solution (including cells) prior to perfusion culture, by tilting the cell culture vessel A4 so that the port 24 is positioned at the top, the air bubbles in the vessel body 1 move along the ceiling surface 231 to the vicinity of the port 24. The air bubbles then rise up through the flow path 242 via the ceiling recess 232. This allows air bubbles remaining in the vessel body 1 to be discharged to the outside of the cell culture vessel A4.

[0098] In addition, the cell culture vessel A4 of this embodiment has the same effects as the cell culture vessel A1 of the first embodiment described above with reference to Figures 1 to 9, etc., within the same range of configuration.

[0099] Although specific embodiments of the present disclosure have been described above, the cell culture vessel according to the present disclosure is not limited to the above-described embodiments. The specific configurations of the components of the cell culture vessel according to the present disclosure can be freely modified in various ways. [Explanation of symbols]

[0100] A1, A11, A12, A13, A14, A2, A3, A4: cell culture vessel, B: air bubbles, C: culture solution, 1: vessel body, 11: first cylindrical portion, 110: opening, 111: male thread (locking means), 112: slit, 113: extension piece, 114: upper end (one side end in the first direction of the first cylindrical portion), 12: vessel bottom, 121: skirt portion, 2: plug member, 2A: first portion, 2B: second portion, 21: second cylindrical portion, 211: annular protrusion, 22: first flange portion, 23: first bottom portion, 231: ceiling surface, 232: ceiling recess, 233: conical portion, 234: peripheral portion, 235: outer peripheral thick portion, 237: outer peripheral edge, 238: thin-walled portion, 24: port, 241: extension portion, 241a: outer peripheral surface, 242: flow path, 243: first flow path, 244: second flow path, 245: protrusion portion, 246: non-penetrating portion, 247: protrusion portion, 26: inner cylindrical portion, 27: upper flange portion, 3: cover member, 31: second flange portion, 32: fourth cylindrical portion, 33: second bottom portion, 331: through hole, 34: fifth cylindrical portion, 4: outer lid, 41: top plate portion, 41a: main surface, 411: through hole, 412: convex portion, 412a: tip, 42: third cylindrical portion, 421: female thread (locking means), 7: connector, 8: tube

Claims

1. a container body having an opening at one end in a first direction and a first cylindrical portion extending in the first direction; a plug member capable of closing the opening, the plug member includes a second cylindrical portion that abuts against and seals an inner circumferential surface of the first cylindrical portion, a first flange portion that extends radially outward from the one side end in the first direction of the second cylindrical portion and closes the one side end in the first direction of the first cylindrical portion, and a plate-shaped first bottom portion that extends radially inward from the other side end in the first direction of the second cylindrical portion and closes the inside of the second cylindrical portion when viewed in the first direction, a plurality of ports are provided in the first bottom portion, each of the ports having a flow path that is open to the outside at one end in the first direction and that is open to the interior space of the container body at the other end in the first direction; the first bottom portion has a ceiling surface facing the other side in the first direction, a ceiling recess formed in the first bottom portion and recessed from the ceiling surface to the one side in the first direction; the ceiling recess is connected to the flow path in at least one of the plurality of ports, the ceiling surface is a flat surface facing the other side in the first direction, the port through which the ceiling recessed portion is connected to the flow path is provided near the outer periphery of the first bottom portion, The cell culture vessel, wherein the ceiling recess communicates with the outer periphery of the first bottom portion.

2. an outer lid including an annular top plate portion and a third cylindrical portion extending from an outer peripheral edge of the top plate portion in a thickness direction of the top plate portion and fitted onto the first cylindrical portion; The cell culture container of claim 1, wherein at least one of the container body and the outer lid is provided with a locking means for preventing relative movement between the first cylindrical portion and the third cylindrical portion when the first flange portion abuts against the first cylindrical portion.

3. 3. The cell culture vessel according to claim 1, wherein the plug member is made of a soft material.

4. The cell culture vessel according to claim 3 , further comprising a cover member made of a hard material and overlapping and abutting the plug member.

5. The cell culture vessel of claim 4, wherein the cover member includes a second flange portion that covers the first flange portion, a fourth cylindrical portion that covers the inner surface of the second cylindrical portion, and a second bottom portion that covers at least a portion of the first bottom portion.

6. the port has an extension extending from the first bottom to the one side in the first direction, The cell culture vessel according to claim 4 , wherein the cover member includes a fifth cylindrical portion that abuts against an outer peripheral surface of the extension portion and surrounds the outer peripheral surface.

7. 4. The cell culture vessel according to claim 3, wherein the first bottom portion includes a thin-walled portion that is relatively thinner than other portions and has gas permeability.

8. the first bottom portion includes a thin-walled portion that is relatively thinner than other portions and has gas permeability; a through-hole penetrating in the first direction is formed in the second bottom portion, The cell culture vessel according to claim 5 , wherein the through-hole overlaps with the thin-walled portion when viewed in the first direction.

9. The cell culture vessel according to claim 6, wherein the tip of the extension portion is provided with a protrusion portion that protrudes outward from the outer circumferential surface and can abut against the one side end of the fifth cylindrical portion in the first direction.

10. the flow path includes a first flow path that is open to the outside, and a second flow path that is open to the internal space of the container body and is connected to the ceiling recess, The cell culture vessel according to claim 6 or 9, wherein the first flow path and the second flow path are spaced apart from each other in the first direction and overlap each other when viewed in the first direction.

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