WO2019182093A1

WO2019182093A1 - Culture vessel

Info

Publication number: WO2019182093A1
Application number: PCT/JP2019/012012
Authority: WO
Inventors: 塚田亮平; 矢ヶ部徹; 大久保春男
Original assignee: 住友ベークライト株式会社
Priority date: 2018-03-23
Filing date: 2019-03-22
Publication date: 2019-09-26
Also published as: JPWO2019182093A1; JP7279703B2

Abstract

A culture vessel is provided which can suitably supply or discharge a culture medium, has excellent sealing performance and which can be easily manufactured. The culture vessel 100 is provided with: a vessel body 2 which has an opening 24A that communicates with a housing space 1; an inner lid 3 which closes the opening 24A; a nozzle 4 which, for supplying or discharging the culture medium 98, is formed integrally with the inner lid 3 in a state passing through the inner lid 3 and extending to the housing space 1; and an outer lid 6 which is provided separately from the inner lid 3 and which fixes the inner lid 3 on the vessel body 2 by pressing.

Description

Culture vessel

The present invention relates to a culture container for culturing cells in a housing space.

In order to culture cells, it is necessary to maintain the growth environment under predetermined conditions. For this reason, a culture container for culturing cells is required to have a sealing property of a storage space in which cells are stored, and a medium needs to be appropriately supplied to and discharged from the storage space. For example, the following Patent Document 1 (Japanese Patent Laid-Open No. 2006-204263) discloses a culture vessel in which the sealing of the accommodation space is ensured and the medium is appropriately supplied and discharged.

In the technique of Patent Document 1, a cap having a female screw is screwed into a neck portion formed in a container body, thereby closing an opening opened at the tip of the neck portion and sealing the accommodation space.

FIG. 7 of Patent Document 1 shows a nozzle formed so as to reach the accommodation space by extending downward from the opening. Thereby, in the technique of patent document 1, the culture medium in the accommodation space can be appropriately discharged. For example, by using such a nozzle for supplying a culture medium, it is possible to appropriately supply the culture medium into the accommodation space.

Here, in the technique of Patent Document 1, a cap for sealing the accommodation space and a nozzle for appropriately discharging (supplying) the culture medium are integrally formed. However, it is difficult to manufacture integrally a fine portion such as a female screw in the cap and a nozzle portion that requires a certain length to reach the accommodation space.

JP 2006-204263 A

There is a need for a culture vessel that can appropriately supply or discharge a medium and that is excellent in hermeticity and can be easily manufactured.

The culture container according to the present invention comprises:
A culture container for culturing cells in the accommodation space having an accommodation space therein,
A container body having an opening communicating with the accommodation space;
An inner lid that closes the opening;
A medium supply or discharge nozzle formed integrally with the inner lid in a state extending through the inner lid to the accommodation space;
An outer lid that is provided separately from the inner lid and presses and fixes the inner lid to the container body.

According to this configuration, since the medium supply or discharge nozzle is formed in a state extending to the accommodation space, the medium can be appropriately supplied to or discharged from the accommodation space. In addition, since the opening is closed by the inner lid integrally formed with the nozzle and the inner lid is pressed and fixed by the outer lid, it is possible to appropriately ensure the tightness of the accommodation space. And the inner cover in which the nozzle for supply or discharge | emission of a culture medium was formed, and the outer cover which can seal an accommodation space appropriately are separate bodies. Therefore, the inner lid and the outer lid can be formed separately, and a culture container that is easy to manufacture as a whole can be realized.

Hereinafter, preferred embodiments of the present invention will be described. However, the scope of the present invention is not limited by the preferred embodiments described below.

As one aspect,
It is mentioned that the inner lid is provided with a gas supply or discharge vent hole penetrating the inner lid.

As one aspect,
It is mentioned that the said container main body has a ceiling part which covers the upper direction of the said accommodation space.

As one aspect,
It is mentioned that the said container main body has a surrounding wall part which covers the circumference | surroundings of the said storage space.

As one aspect,
The container main body has a ceiling part that covers the upper part of the accommodation space and a peripheral wall part that covers the periphery of the accommodation space, and projects from the ceiling part or the peripheral wall part, and its protruding end forms the opening. Part.

As one aspect,
It is mentioned that the outer lid is configured to press and fix the inner lid by rotating the outer lid in a state where the outer lid is sheathed.

As one aspect,
It is mentioned that the nozzle is provided at a position eccentric with respect to the center of the inner lid.

As one aspect,
The container body has a bottom portion that covers a lower portion of the accommodation space,
A cell culture region is provided on the center side of the bottom, and a medium guide portion that is inclined downward toward the cell culture region is formed at the periphery of the cell culture region.

As one aspect,
The container main body has a ceiling part that covers the upper part of the accommodation space, and a cylindrical part that protrudes from the ceiling part and whose protruding end forms the opening part,
The cylindrical part is formed so as to protrude upward from the ceiling part,
It can be mentioned that, when viewed in the vertical direction, the cylindrical portion and the nozzle are arranged at a position overlapping the medium guide portion.

Further features and advantages of the present invention will become more apparent from the following description of exemplary and non-limiting embodiments described with reference to the drawings.

Cross section of culture vessel Vertical view of the culture vessel (plan view) Expanded cross-sectional view of the main part of the culture vessel The principal part expanded sectional view of the culture container concerning other embodiments

An embodiment of a culture vessel will be described with reference to the drawings. As shown in FIG. 1, the culture vessel 100 has an accommodation space 1 inside, and is used for culturing cells 99 in the accommodation space 1. In the accommodation space 1, cells 99 and a medium 98 for providing nutrients to the cells 99 are accommodated. The components of the medium 98 can be appropriately changed according to the purpose of the culture. Moreover, as the culture medium 98, a liquid culture medium, a semi-fluid culture medium, etc. can be used. In the present embodiment, a liquid medium having high fluidity is used as the medium 98.

The culture container 100 includes a container body 2 having an opening 24A communicating with the accommodation space 1, and an inner lid 3 that closes the opening 24A. By closing the opening 24 A with the inner lid 3, it is possible to ensure the sealing property of the accommodation space 1.

As shown in FIGS. 1 and 2, the container body 2 includes a ceiling portion 21 that covers the upper portion of the storage space 1, a plurality of peripheral wall portions 23 that cover the periphery of the storage space 1, and a bottom portion 25 that covers the lower portion of the storage space 1. And have. In the present embodiment, the container body 2 is formed in a rectangular shape when viewed in the vertical direction, and has four peripheral wall portions 23. The accommodation space 1 is formed by being surrounded by a ceiling portion 21, a bottom portion 25, and four peripheral wall portions 23. In addition, as a material of the container body 2, for example, polyethylene or polypropylene can be applied.

The peripheral wall portion 23 is configured to be inclined inward as it goes downward. Therefore, the four peripheral wall portions 23 are configured so as to be constricted inward as they go downward. Thereby, the lower area in the accommodation space 1 can be made narrower, and the lower area in the accommodation space 1 can be easily filled with the medium 98 even with a small amount of the medium 98. Therefore, the culture medium 98 can be easily spread over the cells 99 arranged in the lower region in the accommodation space 1.

The bottom portion 25 is provided with a cell culture region 25A in which the cells 99 are arranged, and a medium guide portion 25B for guiding the medium 98 to the cell culture region 25A. In the illustrated example, the cell culture region 25 A is provided on the center side of the bottom portion 25. And the culture medium guide part 25B is formed so that it may incline below toward the cell culture area | region 25A in the periphery of the cell culture area | region 25A. Thereby, it is possible to appropriately guide the culture medium 98 to the cell culture region 25 A arranged on the center side of the bottom portion 25. In the example shown in FIG. 2, the medium guide portion 25B is formed so as to gradually expand toward the cell culture region 25A when viewed in the vertical direction. Specifically, the culture medium guide portion 25B is formed in a triangular shape (or an arc shape) whose apex is in the vicinity of the position of a supply nozzle 4 to be described later when viewed in the vertical direction. Thereby, the culture medium 98 can be easily spread over the entire cell culture region 25A.

In this embodiment, a surface treatment (for example, a hydrophilization treatment) is performed on a portion corresponding to the cell culture region 25A in the bottom portion 25. Thereby, the cell 99 can be easily adhered to the cell culture region 25A. Such a configuration is suitable when so-called adhesion culture is performed. In the illustrated example, the cell 99 is disposed in the cell culture region 25A by adhering to the cell culture region 25A. However, the culture vessel 100 can also be used when performing so-called suspension culture. In this case, the cell culture region 25A is a region having a predetermined height, and the cells 99 are arranged in the cell culture region 25A in a state of floating in the cell culture region 25A.

The container body 2 has a cylindrical portion 24 that protrudes from the ceiling portion 21 or the peripheral wall portion 23 and whose protruding end forms an opening 24A. In the present embodiment, the cylindrical portion 24 is formed so as to protrude upward from the ceiling portion 21. In the illustrated example, the upper end portion of the tubular portion 24 forms an opening 24 A, and the lower end portion of the tubular portion 24 is connected to the ceiling portion 21. Moreover, in this example, the cylindrical part 24 is formed in the cylindrical shape, and has the external thread part 24S in the outer peripheral surface. Since the opening 24A is formed at the protruding end of the cylindrical portion 24, it is easy to attach the inner lid 3 that closes the opening 24A.

The inner lid 3 closes the opening 24 A formed in the container body 2. As shown in FIG. 3, in the present embodiment, the inner lid 3 has a fitting portion 31 that protrudes toward the inside of the container body 2 in a state where the opening portion 24 A is closed. The fitting portion 31 is configured to abut on the entire circumferential direction of the cylindrical inner peripheral surface 24B formed on the inner surface of the cylindrical portion 24 with the inner lid 3 closing the opening 24A. Yes. A tapered portion 31 A that narrows toward the inner lid center 3 C side toward the distal end side is formed at the distal end portion of the fitting portion 31. The taper portion 31 A has a function of guiding the fitting portion 31 to the opening portion 24 A when the opening portion 24 A is closed by the inner lid 3. In the present embodiment, the inner lid center 3C indicates the center of the inner lid 3 when viewed in the vertical direction (see FIG. 2).

The inner lid 3 has a container side abutting portion 32 that abuts the cylindrical portion 24 (container body 2) from above. The container-side contact portion 32 is formed so as to surround the outer periphery of the fitting portion 31 at the bottom surface portion of the inner lid 3. The inner lid 3 has an outer lid side abutting portion 33 that abuts on the outer lid 6 described later from below. The outer lid side contact portion 33 is formed on the periphery of the upper surface portion of the inner lid 3.

Incidentally, as shown in FIG. 1, in this embodiment, the container body 2 has a pair of openings 24A. One of the pair of openings 24 A is for supplying the culture medium 98 and gas into the storage space 1, and the other is for discharging the culture medium 98 and gas in the storage space 1 from the storage space 1. belongs to. The culture vessel 100 can maintain and maintain the growth environment of the cells 99 in the accommodation space 1 under a predetermined condition by supplying and discharging the medium 98 and gas through the pair of openings 24A. Yes. Therefore, the culture vessel 100 according to the present embodiment is suitable for the case where the supply and discharge of the culture medium 98 are performed without removing various parts while being fixed.

The culture vessel 100 includes a nozzle 4 for supplying or discharging the medium 98 that is integrally formed with the inner lid 3 so as to extend through the inner lid 3 to the accommodation space 1. As a result, the medium 98 can be supplied to or discharged from the storage space 1. As shown in FIG. 1, in this embodiment, the nozzle 4 is provided corresponding to the inner lid 3 that closes each of the pair of openings 24A. One of the pair of nozzles 4 (left nozzle 4 in the illustrated example) is for supplying the medium 98, and the other (right nozzle 4 in the illustrated example) is for discharging the medium 98.

The nozzle 4 protrudes inward and outward of the container body 2 with the inner lid 3 closing the opening 24A. A portion of the nozzle 4 that protrudes inward of the container main body 2 (hereinafter referred to as a nozzle tip portion 41) is disposed in the accommodation space 1. A liquid feed pipe 97 is connected to a portion of the nozzle 4 that protrudes outward from the container body 2 (hereinafter referred to as a nozzle base end portion 42).
The liquid feeding tube 97 is a tube through which the medium 98 flows. In the present embodiment, the medium 98 supplied from the medium supply unit (not shown) to the accommodation space 1 is connected to the liquid supply pipe 97 (the liquid supply pipe 97 on the left side in the figure) connected to the nozzle 4 for supplying the medium 98. Flows through. In addition, the medium 98 discharged from the storage space 1 flows through the liquid feeding pipe 97 (the liquid feeding pipe 97 on the right side in the drawing) connected to the nozzle 4 for discharging the medium 98.

In the present embodiment, the inner lid 3 is provided with a gas supply / discharge vent hole 5 penetrating the inner lid 3. Thereby, supply of the gas with respect to the accommodation space 1 or discharge | emission of the gas from the accommodation space 1 is attained. As shown in FIG. 1, in this embodiment, the vent hole 5 is provided corresponding to the inner lid 3 that closes each of the pair of openings 24A. One of the pair of vent holes 5 (left vent hole 5 in the illustrated example) is for supplying gas, and the other (right vent hole 5 in the illustrated example) is for discharging gas.

In the present embodiment, the vent hole 5 has a pipe connection portion 51 for connecting the air supply pipe 96. The pipe connecting portion 51 protrudes outward from the container body 2 in a state where the inner lid 3 closes the opening 24A. Thereby, the connection of the air supply pipe 96 to the pipe connecting portion 51 is facilitated. In the present embodiment, an air supply pipe 96 (the air supply pipe 96 on the left side in the figure) connected to the pipe connection part 51 in the gas supply vent 5 is supplied from the gas supply part (not shown) to the accommodation space 1. Gas flows. In addition, the gas discharged from the accommodation space 1 flows through the air supply pipe 96 (the air supply pipe 96 on the right side in the drawing) connected to the pipe connection portion 51 in the gas discharge vent 5. However, it is not restricted to such a structure, The vent hole 5 does not need to have the pipe connection part 51. In this case, the vent hole 5 and the air supply pipe 96 are directly connected.

As shown in FIG. 2, in the present embodiment, the cylindrical portion 24 and the nozzle 4 are arranged at a position overlapping the medium guide portion 25 B when viewed in the vertical direction. Thereby, the culture medium 98 can be appropriately supplied with respect to the culture medium guide part 25B, and the culture medium 98 can be appropriately guided with respect to the cell culture region 25A using the inclined surface of the culture medium guide part 25B. In addition, the medium 98 supplied from the nozzle 4 can be prevented from being directly applied to the cells 99 arranged in the cell culture region 25A, and damage of the cells 99 can be suppressed.

In the present embodiment, the nozzle 4 is provided at a position eccentric with respect to the inner lid center 3 C in the inner lid 3. Similarly, the vent hole 5 is also provided at a position eccentric to the inner lid center 3 C in the inner lid 3. Thereby, it is easy to arrange the culture medium 98 and the gas flow path separately in the inner lid 3. Also, for example, it is easy to provide a plurality of nozzles 4 and vent holes 5 for one inner lid 3.

In the present embodiment, the nozzle 4 for discharging the culture medium 98 is eccentric to the center side of the container body 2 with respect to the inner lid center 3C. Here, FIG. 1 shows an example in which the supply nozzle 4 and the discharge nozzle 4 are configured to have the same length. However, as described above, since the discharge nozzle 4 is eccentric to the center side of the container body 2 with respect to the inner lid center 3C, for example, the discharge nozzle 4 is arranged on the inclined surface of the medium guide portion 25B. It becomes possible to extend and arrange | position to the downward direction, avoiding contacting. In other words, the discharge nozzle 4 may be disposed so as to extend further downward than the supply nozzle 4. As a result, the medium 98 stored below the accommodation space 1 can be appropriately discharged.

The culture vessel 100 includes an outer lid 6 that is provided separately from the inner lid 3 and presses and fixes the inner lid 3 against the container body 2. Thereby, the sealing property of the accommodation space 1 can be improved.

In this embodiment, the outer lid 6 has a central opening 61 penetrating in the vertical direction on the center side. The nozzle 4 (specifically, the nozzle base end portion 42) of the inner lid 3 and the pipe connection portion 51 of the vent hole 5 project outward from the container body 2 through the central opening 61. The central opening 61 is formed smaller than the inner lid 3 when viewed in the vertical direction. As a result, the outer lid 6 can press and fix the inner lid 3 from above by another portion while causing the nozzle 4 and the pipe connecting portion 51 to protrude outward from the container body 2 by the central opening 61. ing.

As shown in FIG. 3, in the present embodiment, the outer lid 6 has a pressing portion 62 that presses and fixes the inner lid 3. The pressing portion 62 is configured to face downward with the outer lid 6 being sheathed on the cylindrical portion 24, and contacts the inner lid 3 from above. In the example shown in FIG. 3, the pressing portion 62 is configured to contact the outer lid side contact portion 33 of the inner lid 3.

In the present embodiment, the outer lid 6 is configured to press and fix the inner lid 3 by being rotated in a state where the outer lid 6 is sheathed on the cylindrical portion 24. As shown in FIG. 3, the outer lid 6 has a female screw portion 6 S that meshes with the male screw portion 24 S of the cylindrical portion 24, so that the outer lid 6 can be screwed into the cylindrical portion 24. In a state where the pressing portion 62 of the outer lid 6 and the outer lid side contact portion 33 of the inner lid 3 are in contact with each other, the outer lid 6 is rotated in the screwing direction so that the outer lid side contact portion 33 is moved. Thus, the pressing part 62 can be brought into stronger contact, and the inner lid 3 can be more firmly fixed to the opening 24A (tubular part 24). Thereby, it is possible to further improve the sealing property of the accommodation space 1.

As described above, the culture container 100 includes the inner lid 3 on which the nozzle 4 for supplying or discharging the medium 98 is formed and the outer lid 6 that can appropriately seal the accommodation space 1. Therefore, the inner lid 3 and the outer lid 6 can be formed separately, and the culture container 100 that is easy to manufacture as a whole can be realized.

Furthermore, since the nozzle 4 is provided at a position eccentric with respect to the inner lid center 3 C in the inner lid 3, it is easy to arrange the culture medium 98 and the gas flow path separately in the inner lid 3. . For example, when the nozzle 4 is arranged eccentrically with respect to the inner lid center 3C and the inner lid 3 and the outer lid 6 are integrated, the nozzle 4 is rotated around the inner lid center 3C by a rotation operation at the time of screwing. Also rotate. In this case, there is a high possibility that the rotating nozzle 4 will come into contact with the inclined surface of the medium guide part 25B. ) Is shaved. However, in the present embodiment, as described above, since the inner lid 3 on which the nozzle 4 is formed and the outer lid 6 are separate bodies, the inner lid 3 does not rotate even when the outer lid 6 is rotated. The inner lid 3 can be firmly fixed to the opening 24A (tubular portion 24). Accordingly, the nozzle 4 and the bottom portion 25 (medium guide portion 25B) of the container main body 2 come into contact with each other when the outer lid 6 is screwed while allowing the medium 98 to be appropriately supplied and the storage space 1 to be reliably sealed. Can be suppressed.

[Other Embodiments]
(1) In the above embodiment, the example in which the opening 24A is formed at the protruding end of the cylindrical portion 24 protruding from the ceiling portion 21 of the container body 2 has been described. However, without being limited to such a configuration, as shown in FIG. 4, the opening 24A is directly formed in the ceiling portion 21 of the container body 2, and the container body 2 does not have a cylindrical portion. Also good. In this case, a locking claw 26 for locking the outer lid 6 is formed around the opening 24A, and a locking portion 63 for locking to the locking claw 26 is provided on the outer lid 6. Good to have been. The outer lid 6 locked to the locking claw 26 contacts the inner lid 3 from above and presses and fixes the inner lid 3.

(2) In the above embodiment, an example in which both the nozzle 4 and the vent hole 5 are arranged eccentrically with respect to the inner lid center 3C has been described. However, it is not limited to such a configuration, and either the nozzle 4 or the vent hole 5 may be disposed so as to be located at the inner lid center 3C.

(3) In the above-described embodiment, the example in which the nozzle 4 for supplying the culture medium 98 and the vent hole 5 for supplying gas are separately arranged in the inner lid 3 has been described. However, without being limited to such a configuration, the nozzle 4 may serve both as the supply of the medium 98 and the supply of gas.

(4) In the above embodiment, an example in which the container body 2 has a rectangular shape when viewed in the vertical direction has been described. However, it is not limited to such a configuration, and the shape of the container body 2 when viewed in the vertical direction may be a polygonal shape or a circular shape.

(5) The configuration disclosed in each of the above-described embodiments (including the above-described embodiment and other embodiments; the same applies hereinafter) is applied in combination with the configuration disclosed in the other embodiments unless a contradiction arises. It is also possible to do. Regarding other configurations as well, the embodiments disclosed in the present specification are examples in all respects, and can be appropriately modified without departing from the gist of the present disclosure.

DESCRIPTION OF SYMBOLS 100: Culture container 1: Accommodating space 2: Container body 3: Inner lid 3C: Inner lid center 4: Nozzle 5: Vent 6: Outer lid 21: Ceiling part 23: Peripheral wall part 24: Cylindrical part 24A: Opening part 25 : Bottom 25A: Cell culture region 25B: Medium guide 98: Medium 99: Cells

Claims

A culture container for culturing cells in the accommodation space having an accommodation space therein,
A container body having an opening communicating with the accommodation space;
An inner lid that closes the opening;
A medium supply or discharge nozzle formed integrally with the inner lid in a state extending through the inner lid to the accommodation space;
A culture vessel comprising: an outer lid that is provided separately from the inner lid and presses and fixes the inner lid against the vessel body.
The culture container according to claim 1, wherein a vent for supplying or discharging a gas is provided in the inner lid so as to penetrate the inner lid.
The culture vessel according to claim 1 or 2, wherein the vessel main body has a ceiling portion covering the upper part of the accommodation space.
The culture vessel according to any one of claims 1 to 3, wherein the vessel body has a peripheral wall portion that covers the periphery of the accommodation space.
The container main body has a ceiling part that covers the upper part of the accommodation space and a peripheral wall part that covers the periphery of the accommodation space, and projects from the ceiling part or the peripheral wall part, and its protruding end forms the opening. The culture container according to any one of claims 1 to 4, which has a portion.
The culture container according to claim 5, wherein the outer lid is configured to press and fix the inner lid by rotating the outer lid in a state where the outer lid is sheathed.
The culture vessel according to any one of claims 1 to 6, wherein the nozzle is provided at a position eccentric with respect to a center of the inner lid.
The container body has a bottom portion that covers a lower portion of the accommodation space,
The cell culture region is provided on the center side of the bottom portion, and a medium guide portion that is inclined downward toward the cell culture region is formed at the periphery of the cell culture region. The culture container according to claim 1.
The container main body has a ceiling part that covers the upper part of the accommodation space, and a cylindrical part that protrudes from the ceiling part and whose protruding end forms the opening part,
The cylindrical part is formed so as to protrude upward from the ceiling part,
The culture container according to claim 8, wherein when viewed in the vertical direction, the cylindrical portion and the nozzle are disposed at a position overlapping the medium guide portion.