Culture container and cell culture device

Abstract

Provided is a culture container capable of suppressing bubbling of a culture medium when a liquid culture medium and a gas are supplied together using a common nozzle. The culture container (100) comprises: a container body (2) having an opening (24A) communicating with a storage space (1); a cover (8) sealing the opening (24A); and a nozzle (4) for supplying a liquid culture medium (98) and a gas together, the nozzle (4) penetrating the cover (8) and extending to the storage space (1). The nozzle (4) has a ventilation opening (43), the ventilation opening (43) being formed in a manner of opening in a side wall (45) of the nozzle (4) to form a movement path for the gas from the inside of the nozzle (4) to the storage space (1).

Description

Technical Field

[0001] The invention relates to a culture container and a cell culture device for culturing cells in a containing space. Background Art

[0002] The culture container is supplied with a culture medium (liquid culture medium) that serves as a growth environment for cells and a gas (carbon dioxide) for appropriately maintaining the growth environment. An example of such a culture container is disclosed in the following Patent Document 1 (Japanese Patent Publication No. 2006-204263). Figure 7 As shown, the culture container 30' is provided with an opening 38 connected to a tube 40 for supplying a culture medium B, and the culture medium B is supplied from the opening 38 to the storage space of the culture container 30'. Then, the gas G is supplied from an opening 36 provided at a position different from the opening 38 for supplying the culture medium B.

[0003] Here, from the perspective of reducing the cost of the device and culture container that serve as the supply source of culture medium and gas, technology for supplying culture medium and gas together from one part (opening) of the culture container is studied. For example, it can be considered to use a nozzle extending into the storage space to supply culture medium and gas.

[0004] In this case, when the culture medium is supplied, the culture medium may bubble due to the impact of dripping. In order to suppress this, it is conceivable to supply the culture medium using a nozzle extending near the bottom of the culture container. However, in this case, if the front end of the nozzle is immersed in the liquid surface of the culture medium, the gas is supplied below the liquid surface, so the culture medium will also bubble.

[0005] Prior art literature

[0006] Patent Literature

[0007] Patent Document 1: Japanese Patent Application Publication No. 2006-204263. Summary of the invention

[0008] Problems to be solved by the invention

[0009] There is a demand for a culture container that can suppress the foaming of a culture medium when a liquid culture medium and a gas are supplied together using a common nozzle.

[0010] Means used to solve problems

[0011] The culture container of the present invention is a culture container having a containing space inside and used for culturing cells in the aforementioned containing space, comprising: a container body having an opening connected to the aforementioned containing space; a lid for sealing the aforementioned opening; and a nozzle for supplying a liquid culture medium and a gas together, which penetrates the aforementioned lid and extends to the aforementioned containing space; the aforementioned nozzle has a ventilation opening, and the ventilation opening is formed in a manner of opening on the side wall of the nozzle, forming a movement path of the aforementioned gas from the inside of the nozzle to the aforementioned containing space.

[0012] According to this structure, gas can be supplied to the storage space from the ventilation opening, and as a result, the portion for supplying liquid culture medium (hereinafter, simply referred to as "culture medium") and the portion for supplying gas can be arranged at different positions of the nozzle. Thus, in order to mitigate the impact when the culture medium drips, the portion of the nozzle for supplying the culture medium can be extended to near the bottom of the culture container, and even when the portion for supplying the culture medium is immersed in the liquid surface of the culture medium, the ventilation opening can be arranged above the liquid surface of the culture medium. In this way, according to this structure, a structure that can mitigate the impact when the culture medium drips can be made, and the bubbling of the culture medium caused by the culture medium supplying itself can be suppressed. Moreover, a structure that can supply gas above the liquid surface of the culture medium can be made, and the bubbling of the culture medium caused by the gas supply can be suppressed. Therefore, the bubbling of the culture medium can be suppressed when the culture medium and gas are supplied together using a common nozzle.

[0013] The cell culture device of the present invention is a cell culture device comprising a liquid supply section for supplying a liquid culture medium to the aforementioned receiving space of a culture container for culturing cells in the receiving space, a liquid discharge section for discharging the aforementioned liquid culture medium from the aforementioned receiving space, a gas supply section for supplying a gas to the aforementioned receiving space, an exhaust section for discharging the aforementioned gas from the aforementioned receiving space, and a heat-insulating section for keeping the aforementioned culture container warm, wherein the liquid culture medium supplied from the aforementioned liquid supply section and the gas supplied from the aforementioned gas supply section merge and are supplied together; the aforementioned culture container comprises a container body having an opening section connected to the aforementioned receiving space, a cover for sealing the aforementioned opening section, and a supply nozzle that penetrates the aforementioned cover and extends to the aforementioned receiving space; the aforementioned nozzle has a ventilation opening section, and the ventilation opening section is formed in a manner of opening in the side wall of the nozzle, and serves as a movement path of the aforementioned gas from the inside of the nozzle to the aforementioned receiving space.

[0014] According to this structure, the culture medium and gas can be supplied / discharged at any time in the culture container while the culture container is kept warm, and cells can be automatically cultured in the culture container provided in the cell culture device. At this time, the structure of the device can be simplified by supplying the culture medium supplied from the liquid supply unit and the gas supplied from the gas supply unit together.

[0015] In addition, even when the culture medium and the gas are supplied together from the cell culture device side, the gas can be supplied to the containing space from the ventilation opening in the culture container, and as a result, the part for supplying the culture medium and the part for supplying the gas can be arranged at different positions of the nozzle. Thus, in order to mitigate the impact when the culture medium drips, the part of the nozzle for supplying the culture medium can be extended to the vicinity of the bottom of the culture container, and even when the part for supplying the culture medium is immersed in the liquid surface of the culture medium, the ventilation opening can be arranged above the liquid surface of the culture medium. In this way, according to this structure, in the culture container, a structure that can mitigate the impact when the culture medium drips can be made, and the bubbling of the culture medium caused by the culture medium supplying itself can be suppressed. Moreover, a structure that can supply gas above the liquid surface of the culture medium can be made, and the bubbling of the culture medium caused by the gas supply can be suppressed. Therefore, the bubbling of the culture medium in the culture container can be suppressed while achieving the simplification of the cell culture device.

[0016] Hereinafter, preferred embodiments of the present invention will be described. However, the scope of the present invention is not limited to the preferred embodiments described below.

[0017] As one technical solution, the container body may include a ceiling portion covering an upper portion of the housing space; and the ventilation opening may be formed to include a region above the ceiling portion.

[0018] As one technical solution, it can be mentioned that the ventilation opening is composed of a side wall opening, and the side wall opening is formed continuously or discontinuously from a front end opening provided at the front end of the nozzle.

[0019] As a technical solution, it can be cited that the nozzle has a rib protruding toward the axis of the nozzle on the inner surface of the side wall; the rib is formed across the boundary portion of the formation area where the ventilation opening is formed and the non-formation area where the ventilation opening is not formed in the axial direction of the nozzle.

[0020] As one technical solution, the cap includes an inner cap formed integrally with the nozzle, and an outer cap provided separately from the inner cap and pressing and fixing the inner cap to the container body.

[0021] As one technical solution, the container body may include a ceiling portion covering an upper portion of the housing space and a peripheral wall portion covering a periphery of the housing space.

[0022] As one technical solution, the container body may include a cylindrical portion that protrudes from the ceiling portion or the peripheral wall portion and whose protruding end forms the opening portion.

[0023] As one technical solution, it can be mentioned that the nozzle is provided so as to penetrate the center position of the cover.

[0024] As a technical solution, it can be cited that the aforementioned container body has a bottom covering the lower part of the aforementioned receiving space; a cell culture area is provided on the central side of the aforementioned bottom, and a culture medium guide portion inclined downward toward the aforementioned cell culture area is formed on the periphery of the aforementioned cell culture area.

[0025] As a technical solution, it can be cited that the aforementioned container body has a ceiling portion covering the upper part of the aforementioned containing space and a cylindrical portion protruding from the aforementioned ceiling portion and whose protruding end forms the aforementioned opening portion; the aforementioned cylindrical portion is formed to protrude upward from the aforementioned ceiling portion; when viewed in the up and down directions, the aforementioned cylindrical portion and the aforementioned nozzle are arranged at a position overlapping with the aforementioned culture medium guide portion.

[0026] Further features and advantages of the present invention will become more apparent from the following description of illustrative and non-limiting embodiments with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 is a cross-sectional view of a culture container.

[0028] Figure 2 It is a top-down view of the culture container.

[0029] Figure 3 It is an enlarged cross-sectional view of the main parts showing the periphery of the discharge opening.

[0030] Figure 4 It is an enlarged cross-sectional view of the main parts showing the periphery of the opening for supply.

[0031] Figure 5 It is an enlarged cross-sectional view of the main parts showing the periphery of the opening for supply.

[0032] Figure 6 yes Figure 5 VI-VI section view.

[0033] Figure 7 This is a schematic diagram of a cell culture apparatus.

[0034] Figure 8 This is an enlarged cross-sectional view of the main parts showing the periphery of the supply opening of the culture container according to the second embodiment.

[0035] Fig. 9 yes Figure 8 IX-IX section view.

[0036] Fig.10This is an enlarged cross-sectional view of a main part of a culture container according to another embodiment. DETAILED DESCRIPTION

[0037] [First embodiment]

[0038] A first embodiment of the culture container will be described with reference to the accompanying drawings. Figure 1 As shown, the culture container 100 has a storage space 1 inside, and is used to culture cells 99 in the storage space 1. The cells 99 and a culture medium 98 for providing nutrients to the cells 99 are stored in the storage space 1. The components of the culture medium 98 can be appropriately changed according to the purpose of the culture. In addition, a liquid culture medium (including a semi-fluidized culture medium, etc.) can be used as the culture medium 98. In this embodiment, a liquid culture medium with high fluidity is used as the culture medium 98.

[0039] The culture container 100 includes a container body 2 having an opening 24A communicating with a storage space 1 and a cover 8 that blocks the opening 24A. By blocking the opening 24A with the cover 8, the airtightness of the storage space 1 can be ensured. In this embodiment, the cover 8 includes an inner cover 3 that blocks the opening 24A and an outer cover 6 that is provided separately from the inner cover 3 and presses the inner cover 3 against the container body 2.

[0040] like Figure 1 and Figure 2 As shown, the container body 2 has a ceiling portion 21 covering the upper part of the storage space 1, a plurality of peripheral wall portions 23 covering the periphery of the storage space 1, and a bottom 25 covering the lower part of the storage space 1. In this embodiment, the container body 2 is formed in a rectangular shape when viewed in the up and down direction, and has four peripheral wall portions 23. Moreover, the storage space 1 is surrounded and formed by the ceiling portion 21, the bottom 25 and the four peripheral wall portions 23. In addition, the material of the container body 2, for example, polyethylene or polypropylene can be applied.

[0041] The peripheral wall portion 23 is configured to be inclined inward as it goes downward. Therefore, the four peripheral wall portions 23 are configured as a whole to be narrowed inward as it goes downward. As a result, the lower area in the receiving space 1 can be narrowed, and even with a small amount of culture medium 98, the lower area in the receiving space 1 can be easily filled with the culture medium 98. Therefore, the culture medium 98 can be easily spread over the cells 99 arranged in the lower area of ​​the receiving space 1.

[0042] The bottom portion 25 is provided with a cell culture region 25A where cells 99 are arranged and a culture medium guide portion 25B that guides the culture medium 98 to the cell culture region 25A. In the example shown in the figure, the cell culture region 25A is arranged on the central side of the bottom portion 25. Furthermore, the culture medium guide portion 25B is formed at the periphery of the cell culture region 25A so as to be inclined downward toward the cell culture region 25A. Thus, the culture medium 98 can be appropriately guided relative to the cell culture region 25A arranged on the central side of the bottom portion 25. Figure 2 In the example shown, the culture medium guide 25B is formed so as to gradually expand toward the cell culture area 25A when viewed in the vertical direction. Specifically, the culture medium guide 25B is formed in a triangular shape (more specifically, an isosceles triangle shape) with the cell culture area 25A side as the base when viewed in the vertical direction. Thus, the culture medium 98 can be easily spread throughout the entire cell culture area 25A.

[0043] In the present embodiment, a portion of the bottom 25 corresponding to the cell culture region 25A is subjected to a surface treatment (e.g., a hydrophilic treatment). This allows the cells 99 to be easily adhered to the cell culture region 25A. Such a structure is preferred when so-called adhesion culture is performed. In the example shown in the figure, the cells 99 are arranged in the cell culture region 25A by being adhered to the cell culture region 25A. However, the culture container 100 can also be used when so-called suspension culture is performed. In this case, the cell culture region 25A is an area having a predetermined height, and the cells 99 are arranged in the cell culture region 25A in a state of being suspended in the cell culture region 25A.

[0044] The container body 2 has a cylindrical portion 24 protruding from the ceiling portion 21 or the peripheral wall portion 23 and having an opening 24A formed at its protruding end. 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 cylindrical portion 24 forms the opening 24A, and the lower end portion of the cylindrical portion 24 is connected to the ceiling portion 21. In addition, in the present example, the cylindrical portion 24 is formed in a cylindrical shape and has a male thread portion 24S on its outer circumferential surface. Since the opening 24A is formed at the protruding end of the cylindrical portion 24, the installation of the inner cover 3 that blocks the opening 24A becomes easy.

[0045] like Figure 1As shown, in the present embodiment, the container body 2 has a pair of openings 24A. Moreover, one of the pair of openings 24A is used to supply the culture medium 98 and gas into the receiving space 1, and the other is used to discharge the culture medium 98 and gas in the receiving space 1 from the receiving space 1. The culture container 100 can maintain the growth environment of the cells 99 in the receiving space 1 under prescribed conditions by supplying and discharging the culture medium 98 and gas through the pair of openings 24A. Therefore, the culture container 100 related to the present embodiment is preferably used in a case where the culture medium 98 is supplied and discharged while maintaining the various parts in a fixed state without being disassembled. In the following description, the opening 24A ( Figure 1 The opening 24A on the left side shown in the figure is referred to as the "supply opening 24A". The opening 24A ( Figure 1 The opening 24A on the right side shown is referred to as the "discharge opening 24A".

[0046] First, refer to Figure 1 to Figure 3 The structure around the discharge opening 24A will be described.

[0047] The inner cover 3 closes the opening 24A formed in the container body 2. Figure 3 As shown, in the present embodiment, the inner cover 3 has a fitting portion 31 that protrudes toward the inside of the container body 2 when the opening portion 24A is blocked. The fitting portion 31 is configured to abut the entire circumference of the cylindrical inner peripheral surface 24B formed on the inner surface of the cylindrical portion 24 when the inner cover 3 blocks the opening portion 24A. At the front end portion of the fitting portion 31, a tapered portion 31A is formed that narrows toward the inner cover center 3C side as it moves toward the front end side. The tapered portion 31A has a function of guiding the fitting portion 31 toward the opening portion 24A when the opening portion 24A is blocked by the inner cover 3. In addition, in the present embodiment, the inner cover center 3C refers to the center of the inner cover 3 as viewed in the up and down direction (refer to Figure 2 ).

[0048] The inner cover 3 has a container side contact portion 32 that contacts the cylindrical portion 24 (container body 2) from above. The container side contact portion 32 is formed on the bottom surface portion of the inner cover 3 so as to surround the outer periphery of the fitting portion 31. In addition, the inner cover 3 has an outer cover side contact portion 33 that contacts the outer cover 6 described later from below. The outer cover side contact portion 33 is formed on the periphery of the upper surface portion of the inner cover 3.

[0049] The culture container 100 includes a discharge nozzle 7 for discharging gas that penetrates the cover 8 and extends to the housing space 1. In the present embodiment, the discharge nozzle 7 penetrates the inner cover 3 that blocks the discharge opening 24A and extends to the housing space 1. The inner cover 3 that blocks the discharge opening 24A and the discharge nozzle 7 are formed integrally.

[0050] The discharge nozzle 7 protrudes inward and outward from the container body 2 when the inner cover 3 blocks the discharge opening 24A. The portion of the discharge nozzle 7 protruding inward from the container body 2 is disposed in the storage space 1. The portion of the discharge nozzle 7 protruding outward from the container body 2 is connected to a liquid delivery tube 97. The liquid delivery tube 97 is a tube through which the culture medium 98 flows. In the present embodiment, the culture medium 98 discharged from the storage space 1 flows through the liquid delivery tube 97.

[0051] like Figure 2 As shown in FIG. 1 , in this embodiment, the discharge nozzle 7 is disposed at a position eccentric to the inner cap center 3C of the inner cap 3. The discharge nozzle 7 is eccentric to the center side of the container body 2 relative to the inner cap center 3C. Figure 1 , an example in which the discharge nozzle 7 and the supply nozzle 4 described later are configured to have the same length is shown. However, by eccentrically positioning the discharge nozzle 7 toward the center of the container body 2 relative to the inner cover center 3C as described above, for example, the discharge nozzle 7 can be extended further downward while avoiding contact with the inclined surface of the culture medium guide 25B. In other words, the discharge nozzle 7 can also be extended downward than the supply nozzle 4. Thus, the culture medium 98 accumulated at the bottom of the storage space 1 can be appropriately discharged.

[0052] like Figure 1 As shown, in the present embodiment, the inner cover 3 that blocks the opening 24A for discharge is provided with a vent hole 5 for gas discharge in a state that the inner cover 3 is penetrated. Thus, the gas from the accommodation space 1 can be discharged. In the example shown in the figure, the vent hole 5 is provided at a position eccentric to the inner cover center 3C of the inner cover 3. In this way, by eccentrically arranging the discharge nozzle 7 and the vent hole 5 relative to the inner cover center 3C, it is easy to separately arrange the flow path of the culture medium 98 and the gas in the inner cover 3. In addition, for example, it is also easy to provide a plurality of discharge nozzles 7 and vent holes 5 described later with respect to one inner cover 3.

[0053] In the present embodiment, the vent hole 5 has a tube connection portion 51 for connecting the air supply pipe 96. The tube connection portion 51 protrudes toward the outside of the container body 2 when the inner cover 3 blocks the opening 24A for discharge. As a result, it is easy to connect the air supply pipe 96 to the tube connection portion 51. The gas discharged from the accommodation space 1 flows through the air supply pipe 96. However, the vent hole 5 is not limited to the above-mentioned structure, and the vent hole 5 may not have the tube connection portion 51. In this case, the vent hole 5 and the air supply pipe 96 are directly connected.

[0054] As described above, the culture container 100 includes an outer cover 6 which is provided separately from the inner cover 3 and which presses and fixes the inner cover 3 relative to the container body 2. Thus, the airtightness of the storage space 1 can be improved. In the present embodiment, the outer cover 6 has a central opening portion 61 which penetrates in the up-down direction on its central side. The discharge nozzle 7 and the tube connection portion 51 protrude to the outside of the container body 2 through the central opening portion 61. When viewed in the up-down direction, the central opening portion 61 is formed to be smaller than the inner cover 3. Thus, the outer cover 6 can press and fix the inner cover 3 from above by means of other parts while making the discharge nozzle 7 and the tube connection portion 51 protrude to the outside of the container body 2 by means of the central opening portion 61.

[0055] like Figure 3 As shown, in this embodiment, the outer cover 6 has a pressing portion 62 for pressing and fixing the inner cover 3. The pressing portion 62 is configured to face downward when the outer cover 6 is attached to the cylindrical portion 24, and to abut against the inner cover 3 from above. Figure 3 In the illustrated example, the pressing portion 62 is configured to come into contact with the outer cover side contact portion 33 of the inner cover 3 .

[0056] In the present embodiment, the outer cover 6 is configured to be rotated while being mounted on the cylindrical portion 24, thereby pressing and fixing the inner cover 3. Figure 3 As shown, the outer cover 6 has a female thread portion 6S that meshes with the male thread portion 24S of the cylindrical portion 24, thereby being able to be threadedly engaged with the cylindrical portion 24. By rotating the outer cover 6 in the threaded engagement direction while the pressing portion 62 of the outer cover 6 is in contact with the outer cover side contact portion 33 of the inner cover 3, the pressing portion 62 can be more strongly contacted with the outer cover side contact portion 33, and the inner cover 3 can be more firmly fixed to the opening portion 24A (cylindrical portion 24). As a result, the airtightness of the accommodating space 1 can be further improved.

[0057] As described above, the culture container 100 has the inner cover 3 and the outer cover 6 that can appropriately seal the storage space 1 as separate bodies. Therefore, the inner cover 3 and the outer cover 6 can be formed separately, and the culture container 100 that is easy to manufacture as a whole can be realized.

[0058] Furthermore, since the discharge nozzle 7 is provided at a position eccentric to the inner cover center 3C of the inner cover 3 , it is easy to separately arrange the flow paths of the culture medium 98 and the gas in the inner cover 3 .

[0059] For example, when the discharge nozzle 7 is eccentrically arranged relative to the inner cover center 3C and the inner cover 3 and the outer cover 6 are integrated, the discharge nozzle 7 also rotates around the inner cover center 3C due to the rotation operation during the screw connection. In this case, the possibility of the rotating discharge nozzle 7 contacting the inclined surface of the culture medium guide portion 25B becomes high, and in the case of contact, the discharge nozzle 7 is damaged or the bottom 25 (culture medium guide portion 25B) of the container body 2 is cut off. However, in this embodiment, as described above, the inner cover 3 and the outer cover 6 formed with the discharge nozzle 7 are separate bodies, so the inner cover 3 will not rotate due to the rotation operation of the outer cover 6, and the inner cover 3 can be firmly fixed to the opening 24A (cylindrical portion 24). As a result, while the appropriate discharge of the culture medium 98 and the reliable sealing of the storage space 1 can be performed, the discharge nozzle 7 can be prevented from contacting the bottom 25 (culture medium guide portion 25B) of the container body 2 during the screw connection of the outer cover 6.

[0060] Next, refer to Figure 1 , Figure 2 and Figures 4 to 6 The structure around the supply opening 24A will be described. Since the structures of the cap 8, the cylindrical portion 24, etc. are substantially the same as the structure around the discharge opening 24A described above, the following description will focus on the differences.

[0061] The culture container 100 includes a supply nozzle 4 for supplying a culture medium 98 and a gas, which extends through the cover 8 to the storage space 1. The supply nozzle 4 is a common nozzle for supplying both the culture medium 98 and the gas. In the present embodiment, the supply nozzle 4 corresponds to a "nozzle".

[0062] In the present embodiment, the supply nozzle 4 passes through the inner cover 3 that blocks the opening 24A for supply and extends to the housing space 1. The inner cover 3 that blocks the opening 24A for supply and the supply nozzle 4 are formed integrally.

[0063] In the present embodiment, the supply nozzle 4 is provided so as to penetrate the center position of the cap 8 .

[0064] exist Figure 2 In the example shown, the supply nozzle 4 is provided to penetrate the inner cap center 3C of the inner cap 3. The supply nozzle 4 is formed in a cylindrical shape and arranged so that the axis AX of the supply nozzle 4 overlaps the inner cap center 3C when viewed in the vertical direction.

[0065] The supply nozzle 4 protrudes toward the inside and outside of the container body 2 when the inner cover 3 blocks the opening 24A. In the present embodiment, the supply nozzle 4 has an inner front end portion 41 (first front end portion) as the front end portion on the inner side of the container body 2 of the supply nozzle 4 and an outer front end portion 42 (second front end portion) as the front end portion on the outer side of the container body 2 of the supply nozzle 4. The inner front end portion 41 is arranged in the storage space 1, and the outer front end portion 42 is arranged outside the storage space 1. The liquid supply and gas supply pipe 95 is connected to the outer front end portion 42. The liquid supply and gas supply pipe 95 is a pipe through which the culture medium 98 and the gas flow. In the present embodiment, the culture medium 98 supplied to the storage space 1 from the culture medium supply portion not shown and the gas supplied to the storage space 1 from the gas supply portion not shown flow through the liquid supply and gas supply pipe 95 at the same time.

[0066] The supply nozzle 4 has a front end opening portion 41A which is open at its front end and communicates with the storage space 1. In the present embodiment, the front end opening portion 41A is formed at the inner front end portion 41 of the supply nozzle 4. The culture medium 98 flowing inside the supply nozzle 4 is supplied into the storage space 1 via the front end opening portion 41A. The front end opening portion 41A is a "liquid passage opening portion" which mainly serves as a movement path of the liquid culture medium.

[0067] like Figure 2 As shown, in this embodiment, the cylindrical portion 24 and the supply nozzle 4 are arranged at a position overlapping with the culture medium guide portion 25B when viewed in the vertical direction. Thus, the culture medium 98 can be appropriately supplied to the culture medium guide portion 25B, and the culture medium 98 can be appropriately guided relative to the cell culture area 25A using the inclined surface of the culture medium guide portion 25B. In addition, the culture medium 98 supplied from the supply nozzle 4 can be prevented from directly acting on the cells 99 arranged in the cell culture area 25A, and damage to the cells 99 can be suppressed.

[0068] like Figure 5 and Figure 6 As shown, the supply nozzle 4 has a ventilation opening portion (side wall opening portion) 43 formed in a manner of opening on the side wall 45 of the supply nozzle 4. The ventilation opening portion 43 is a "side opening portion" formed on the side wall 45 of the supply nozzle 4 in a manner of facing to the side. The ventilation opening portion 43 becomes a movement path for the gas from the inside of the supply nozzle 4 to the storage space 1. Thus, the part for supplying the culture medium 98 and the part for supplying the gas can be arranged at different parts of the supply nozzle 4. In the present embodiment, the culture medium 98 is supplied from the front end opening portion 41A (liquid passage opening portion) of the supply nozzle 4, and the gas is supplied from the side opening portion (ventilation opening portion 43) of the supply nozzle 4.

[0069] like Figure 5As shown, in the present embodiment, the ventilation opening 43 is formed to include an area above the ceiling 21. In other words, the upper end of the ventilation opening 43 is arranged above the ceiling 21. Thus, for example, even when the storage space 1 is filled with the culture medium 98 up to the vicinity of the ceiling 21, gas can be supplied above the liquid surface of the culture medium 98.

[0070] In the present embodiment, the ventilation opening 43 is formed by a slit formed continuously from the front end opening 41A provided at the inner front end portion 41 of the supply nozzle 4. The ventilation opening 43 formed by the slit is formed along the axis AX of the supply nozzle 4. According to this structure, when manufacturing the culture container 100, it is easy to form the ventilation opening 43 on the side wall 45 of the supply nozzle 4, which can make the manufacturing easy. The ventilation opening 43 is preferably formed over a range of 1 / 3 to 2 / 3 of the axial length of the supply nozzle 4. According to this structure, while the entire ventilation opening 43 is not immersed in the liquid surface of the culture medium 98, it is easy to ensure the durability of the supply nozzle 4.

[0071] like Figure 6 As shown, in the present embodiment, the ventilation opening 43 is formed in a manner of opening on both sides of the axis AX of the supply nozzle 4. Thus, the movement path of the gas to the storage space 1 can be ensured to be larger, and the gas flowing inside the supply nozzle 4 can be supplied to the storage space 1 before reaching the front end opening 41A. However, it is not limited to such a structure, and the ventilation opening 43 can also be formed at one place in the circumferential direction of the supply nozzle 4 when viewed from the axial direction.

[0072] As described above, the supply nozzle 4 has a ventilation opening 43 formed in a manner of opening in the side wall 45 of the supply nozzle 4 and serving as a moving path for gas from the inside of the supply nozzle 4 to the storage space 1. Therefore, the part for supplying the culture medium 98 and the part for supplying the gas can be arranged at different parts of the supply nozzle 4.

[0073] Furthermore, the inner front end portion 41 for supplying the culture medium 98 is formed at the front end of the supply nozzle 4, and the ventilation opening portion 43 for supplying the gas is formed to include an area above the ceiling portion 21. Therefore, the impact of the culture medium 98 when dripping can be mitigated, and the gas can be supplied above the liquid surface of the culture medium 98. Therefore, the bubbling of the culture medium 98 when the culture medium 98 and the gas are supplied can be suppressed.

[0074] In this embodiment, as described above, the supply nozzle 4 supplies the culture medium 98 and the gas together from one opening of the storage space 1. The culture medium 98 and the gas are discharged from the storage space 1 independently by the gas vent 5 and the liquid discharge nozzle 7. Figure 71 is a diagram showing a structural example of a cell culture apparatus for automatically performing cell culture using the culture container 100. The liquid supply unit 71 is a liquid supply pump for supplying liquid to the storage space 1, and is connected to a liquid bottle (not shown) at its upstream, and is connected to a connection unit 73 at its downstream via a liquid supply control valve 72. The gas supply unit 74 is a gas supply source for supplying gas to the storage space 1, and is connected to the connection unit 73 via a gas supply control valve 75, and the connection unit 73 is connected to the supply nozzle 4 via a liquid and gas supply pipe 95.

[0075] The liquid discharge unit 77 is a liquid discharge pump for discharging liquid from the storage space 1. The upstream connected by the liquid supply pipe 97 is connected to the discharge nozzle 7 via the liquid discharge control valve 79, and the downstream is connected to the liquid discharge bottle (not shown). The air supply pipe 96 is an exhaust pipe for discharging gas connected to the vent hole 5 of the culture container 100, and is connected to the ventilation filter 82 via the exhaust control valve 81, and the downstream is opened to the atmosphere.

[0076] In this embodiment, when air is supplied to the storage space 1, the air supply control valve 75 and the exhaust control valve 81 are opened, the liquid supply control valve 72 and the drain control valve 79 are kept closed, and air is supplied at a predetermined air supply amount from the air supply portion 74. Then, the supplied gas passes through the air supply control valve 75, is introduced from the supply nozzle 4 to the storage space 1 via the ventilation opening 43, and is released from the air supply pipe 96 through the exhaust control valve 81 and the ventilation filter 82 to the atmosphere.

[0077] When liquid is supplied to the storage space 1, the liquid supply control valve 72 and the exhaust control valve 81 are opened, and a predetermined liquid supply process is performed from the liquid supply section 71. Thus, the gas initially held inside the liquid supply and air supply pipe 95 is introduced into the storage space 1 from the supply nozzle 4 through the ventilation opening 43, and then the desired liquid is introduced into the storage space 1 from the liquid bottle not shown. Then, the remaining gas is pressed out, and the desired liquid is delivered by means of the liquid supply section 71. At this time, as described above, since the exhaust control valve 81 is opened, the gas equivalent to the sum of the introduced gas and liquid is exhausted from the ventilation filter 82.

[0078] When draining liquid from the storage space 1, the gas supply control valve 75 and the liquid discharge control valve 79 are opened, and a predetermined liquid discharge process is performed from the liquid discharge section 77, whereby the gas initially held inside the liquid supply pipe 97 is exhausted to a liquid discharge bottle (not shown) through the liquid discharge control valve 79. Then, the culture medium 98 is sucked from the discharge nozzle 7, flows through the liquid supply pipe 97, passes through the liquid discharge control valve 79, and is discharged to the liquid discharge bottle. On the other hand, at this time, the gas supply control valve 75 is opened, and gas can be supplied from the gas supply section 74, and gas equivalent to the sum of the exhausted gas and liquid is supplied from the gas supply section 74, so that the inside of the storage space 1 can be filled.

[0079] The cell culture device of this embodiment includes a heat preservation part (not shown) for keeping the culture container 100 warm. The heat preservation part is preferably composed of a constant temperature part (constant temperature box (incubator)) for keeping the culture container 100 at a certain temperature. By keeping the culture container 100 at a temperature suitable for cell culture by the heat preservation part (constant temperature box), cell culture can be performed. In this way, the cell culture device includes a liquid supply part 71 for supplying liquid such as a culture medium 98 to the culture container 100, a liquid discharge part 77 for discharging liquid such as the culture medium 98, a gas supply part 74 for supplying gas for ventilation, and a gas exhaust part (gas supply pipe 96) for exhausting gas. By providing a culture container 100 that suppresses the bubbling of the culture medium 98 when the culture medium 98 and gas are supplied together, and keeping the culture container 100 warm by the heat preservation part (constant temperature box), cell culture can be automatically performed.

[0080] [Second embodiment]

[0081] Next, refer to Figure 8 and Fig. 9 A second embodiment of the culture container 100 will be described. Hereinafter, differences from the first embodiment will be mainly described. Points not particularly described are the same as those of the first embodiment.

[0082] like Figure 8 and Fig. 9 As shown in FIG. 1 , the supply nozzle 4 has a rib 44 protruding toward the axis AX of the supply nozzle 4 on the inner surface of the side wall 45 of the supply nozzle 4. The culture medium 98 flowing down the inside of the supply nozzle 4 flows along the rib 44 by the action of the surface tension generated between the culture medium 98 and the rib 44. With the help of the rib 44, the culture medium 98 can flow inside the supply nozzle 4 in a manner avoiding the ventilation opening 43, and the culture medium 98 can smoothly reach the front end opening 41A. In addition, since the ventilation opening 43 becomes less likely to be blocked by the culture medium 98, the gas supply from the ventilation opening 43 can be smoothly performed.

[0083] like Fig. 9 As shown, the rib 44 and the ventilation opening 43 are formed at different positions in the circumferential direction based on the axis AX of the supply nozzle 4. In the example shown in the figure, the rib 44 is formed in a pair on both sides of the axis AX at a position that is 90° phase-shifted relative to the ventilation opening 43 formed in a manner of opening on both sides of the axis AX. However, it is not limited to such a structure, and the rib 44 can also be formed at one position in the circumferential direction of the supply nozzle 4 when viewed from the axial direction.

[0084] like Figure 8As shown, the rib 44 is formed across a boundary portion 43A between a formation area where the ventilation opening portion 43 is formed and a non-formation area where the ventilation opening portion 43 is not formed in the axial direction of the supply nozzle 4. In other words, the rib 44 is formed across the upstream end portion of the ventilation opening portion 43 in the gas flow direction. With such a structure, the culture medium 98 can be properly guided toward the front end opening portion 41A while allowing the gas to move from the ventilation opening portion 43 into the storage space 1. In the present embodiment, the rib 44 is formed from the non-formation area where the ventilation opening portion 43 is not formed to the front end opening portion 41A. As a result, the culture medium 98 can be guided toward the front end opening portion 41A with high reliability.

[0085] [Other implementation methods]

[0086] (1) In the above-mentioned embodiment, an example is described in which the ventilation opening portion 43 is composed of a slit formed continuously from the front end opening portion 41A provided at the inner front end portion 41 of the supply nozzle 4. However, it is not limited to such a structure, and the ventilation opening portion 43 may be formed, for example, over a range of more than 2 / 3 of the axial length, or the supply nozzle 4 may be formed at more than 3 locations in the circumferential direction when viewed from the axial direction. Furthermore, the ventilation opening portion 43 may be formed at a position away from the front end opening portion 41A (in other words, as an opening portion independent of the front end opening portion 41A). For example, the ventilation opening portion 43 may also be composed of a hole portion that penetrates the side wall 45 of the supply nozzle 4. In this case, the ventilation opening portion 43 may also be composed of a plurality of hole portions. In addition, the rib portion 44 is not limited to the structure of the above-mentioned embodiment, and for example, the supply nozzle 4 may be formed at more than 3 locations in the circumferential direction when viewed from the axial direction.

[0087] (2) In the above-mentioned 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 is described. However, the present invention is not limited to such a structure, and the opening 24A may be formed at the protruding end of the cylindrical portion 24 protruding from the ceiling portion 21 of the container body 2. Fig.10 As shown, the opening 24A is directly formed on the ceiling 21 of the container body 2, and the container body 2 has a structure without a cylindrical portion. In this case, a locking claw 26 for locking the outer cover 6 can be formed around the opening 24A, and a locking portion 63 for locking with the locking claw 26 can be provided on the outer cover 6. The outer cover 6 locked by the locking claw 26 abuts against the inner cover 3 from above, and the inner cover 3 is pressed and fixed.

[0088] (3) In the above embodiment, the discharge nozzle 7 and the vent hole 5 are arranged eccentrically with respect to the inner cap center 3C. However, the present invention is not limited to such a structure, and either the discharge nozzle 7 or the vent hole 5 may be arranged so as to be located at the inner cap center 3C.

[0089] (4) In the above-described embodiment, an example is described in which the cap 8 that blocks the opening 24A for supply includes the inner cap 3 that integrally includes the supply nozzle 4 and the outer cap 6 that is separate from the inner cap 3. However, the present invention is not limited to such a structure, and the cap 8 may be configured as an integral body that includes the supply nozzle 4. The same can be considered with respect to the cap 8 that blocks the opening 24A for discharge.

[0090] (5) In the above embodiment, the container body 2 is described as being rectangular when viewed in the vertical direction. However, the present invention is not limited to such a configuration, and the container body 2 may be polygonal or circular when viewed in the vertical direction.

[0091] (6) The structures disclosed in the above-mentioned embodiments (including the above-mentioned embodiments and other embodiments; the same applies hereinafter) may be combined and applied with structures disclosed in other embodiments as long as no contradiction occurs. Regarding other structures, the embodiments disclosed in this specification are illustrative in all points and can be appropriately changed within the scope of the present disclosure.

[0092] Description of Reference Numerals

[0093] 100: Culture container

[0094] 1: Containment space

[0095] 2: Container body

[0096] 3: Inner cover

[0097] 3C: Inner cover center

[0098] 4: Supply nozzle (nozzle)

[0099] 6: Outer cover

[0100] 8: Cover

[0101] 21: Ceiling

[0102] 23: Peripheral wall

[0103] 24: Cylindrical part

[0104] 24A: Opening

[0105] 25: Bottom

[0106] 25A: Cell culture area

[0107] 25B: Culture medium guide

[0108] 41: Inner front end (front end of the nozzle)

[0109] 41A: Front opening

[0110] 43: Ventilation opening

[0111] 44: Ribs

[0112] 45: Sidewall

[0113] 98: Culture medium

[0114] 99: Cells

[0115] AX: axis.

Claims

1. A culture container having a receiving space inside, used for culturing cells in the receiving space, characterized in that: have: A container body having an opening communicating with the aforementioned receiving space; A cover to seal the opening; and A nozzle for supplying both the liquid culture medium and the gas, penetrating the cover and extending to the housing space; The nozzle has a front end opening and a ventilation opening. The front end opening is opened at the front end of the nozzle and communicates with the housing space. The ventilation opening is formed in a manner of opening on the side wall of the nozzle to form a movement path for the gas from the inside of the nozzle to the housing space. The liquid culture medium is supplied to the housing space via the front end opening. The front end opening is extended to the vicinity of the bottom of the culture container. The ventilation opening is arranged above the liquid surface of the liquid culture medium.

2. The culture container according to claim 1, characterized in that The container body has a ceiling portion covering the upper portion of the accommodation space; The ventilation opening is formed to include a region above the ceiling.

3. The culture container according to claim 1 or 2, characterized in that: The ventilation opening is composed of a side wall opening formed continuously or discontinuously from a front end opening provided at a front end portion of the nozzle.

4. The culture container according to any one of claims 1 to 3, characterized in that The nozzle has a rib protruding toward the axis of the nozzle on the inner surface of the side wall; The rib portion is formed across a boundary portion between a formation region where the ventilation opening portion is formed and a non-formation region where the ventilation opening portion is not formed in the axial direction of the nozzle.

5. The culture container according to any one of claims 1 to 4, characterized in that The cap includes an inner cap formed integrally with the nozzle and an outer cap provided separately from the inner cap and pressing and fixing the inner cap to the container body.

6. The culture container according to any one of claims 1 to 5, characterized in that The container body includes a ceiling portion covering the upper side of the housing space and a peripheral wall portion covering the periphery of the housing space.

7. The culture container according to claim 6, characterized in that The container body has a cylindrical portion that protrudes from the ceiling portion or the peripheral wall portion and whose protruding end forms the opening portion.

8. The culture container according to any one of claims 1 to 7, wherein The nozzle is provided so as to penetrate the center position of the cover.

9. The culture container according to any one of claims 1 to 8, wherein The container body has a bottom covering the lower part of the receiving space; A cell culture area is provided at the center of the bottom portion, and a culture medium guide portion is formed at the periphery of the cell culture area and is inclined downward toward the cell culture area.

10. The culture container according to claim 9, characterized in that The container body comprises a ceiling portion covering the upper side of the storage space and a cylindrical portion protruding from the ceiling portion and having a protruding end forming the opening; The cylindrical portion is formed to protrude upward from the ceiling portion; The cylindrical portion and the nozzle are arranged at a position overlapping with the culture medium guide portion when viewed in the up-down direction.

11. A cell culture device comprising a liquid supply unit for supplying a liquid culture medium to the storage space of a culture container for culturing cells in the storage space, a liquid discharge unit for discharging the liquid culture medium from the storage space, a gas supply unit for supplying a gas to the storage space, a gas exhaust unit for discharging the gas from the storage space, and a heat preservation unit for keeping the culture container warm, characterized in that: The liquid culture medium supplied from the liquid supply unit and the gas supplied from the gas supply unit merge and are supplied together; The culture container comprises a container body having an opening communicating with the housing space, a cover for closing the opening, and a supply nozzle penetrating the cover and extending to the housing space; The nozzle has a front end opening and a ventilation opening. The front end opening is opened at the front end of the nozzle and communicates with the housing space. The ventilation opening is formed in a manner of opening on the side wall of the nozzle to form a movement path for the gas from the inside of the nozzle to the housing space. The liquid culture medium is supplied to the housing space via the front end opening. The front end opening is extended to the vicinity of the bottom of the culture container. The ventilation opening is arranged above the liquid surface of the liquid culture medium.

12. The cell culture device according to claim 11, characterized in that: The container body has a ceiling portion covering the upper portion of the accommodation space; The ventilation opening is formed to include a region above the ceiling.

13. The cell culture device according to claim 11 or 12, characterized in that: The ventilation opening is composed of a side wall opening formed continuously or discontinuously from a front end opening provided at a front end portion of the nozzle.

14. The cell culture device according to any one of claims 11 to 13, wherein: The nozzle has a rib protruding toward the axis of the nozzle on the inner surface of the side wall; The rib portion is formed across a boundary portion between a formation region where the ventilation opening portion is formed and a non-formation region where the ventilation opening portion is not formed in the axial direction of the nozzle.

15. The cell culture device according to any one of claims 11 to 14, wherein: The cap includes an inner cap formed integrally with the nozzle and an outer cap provided separately from the inner cap and pressing and fixing the inner cap to the container body.

16. The cell culture device according to any one of claims 11 to 15, wherein: The container body includes a ceiling portion covering the upper side of the housing space and a peripheral wall portion covering the periphery of the housing space.

17. The cell culture device according to claim 16, wherein: The container body has a cylindrical portion that protrudes from the ceiling portion or the peripheral wall portion and whose protruding end forms the opening portion.

18. The cell culture device according to any one of claims 11 to 17, wherein: The nozzle is provided so as to penetrate the center position of the cover.

19. The cell culture device according to any one of claims 11 to 18, wherein: The container body has a bottom covering the lower part of the receiving space; A cell culture area is provided at the center of the bottom portion, and a culture medium guide portion is formed at the periphery of the cell culture area and is inclined downward toward the cell culture area.

20. The cell culture device according to claim 19, wherein: The container body comprises a ceiling portion covering the upper side of the storage space and a cylindrical portion protruding from the ceiling portion and having a protruding end forming the opening; The cylindrical portion is formed to protrude upward from the ceiling portion; The cylindrical portion and the nozzle are arranged at a position overlapping with the culture medium guide portion when viewed in the up-down direction.

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