Bag holding clip

Abstract

To provide a bag gripping clip and a stirring method which can prevent a decrease in cell recovery rate in the process of stirring a cell suspension.SOLUTION: The present invention provides a bag gripping clip, wherein, in a case where a bag 2 that forms an accommodation space between a first film and a second film facing the first film is gripped by the clip, a pressing force for bringing the first film and the second film into partially close contact with each other is applied to the bag, and a partition 30 that divides the accommodation space into a first space 31 and a second space 32 communicating with each other via a communication part 33 is formed. The bag gripping clip is attachable to and detachable from the bag. In the bag gripping clip, the partition includes a section extending in a cross direction crossing a first side A1 and a second side A2, facing the first side, of the bag between the first side and the second side; and the communication part is provided between a section extending from the first side of the partition toward the second side and a section extending from the second side of the partition toward the first side.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] The disclosed technology relates to a bag gripping clip, a container, and a stirring method. [Background technology]

[0002] The following technologies are known as technologies related to cell culture bags. For example, Japanese Patent Application Laid-Open No. 2011-239734 describes a technology in which a culture vessel is divided into two chambers, a culture section and an expandable section, by a partition member, and the volume of the culture section is expanded by moving the position of the partition member as the number of cells increases. Summary of the Invention [Problem to be solved by the invention]

[0003] In suspension culture, in which cells are cultured in a suspended state in a medium, it is necessary to distribute nutrients and gases necessary for cell growth to all cells, so it is preferable that the cells are uniformly dispersed in the medium.

[0004] A method for uniformly dispersing cells in a culture medium is to stir the cell suspension using a stirring blade, but cells are fragile and may be damaged by the physical stress caused by the stirring blade, resulting in a decrease in cell viability.

[0005] Another method for uniformly dispersing cells in a medium is known as rocking agitation, in which a bag containing cells and medium is rocked. However, rocking agitation causes more drastic changes in the flow state of the cell suspension than agitation using a stirring blade, making it difficult to establish operating conditions.

[0006] Another method for uniformly dispersing cells in a medium is to apply pressure to the cell suspension contained in a bag to cause it to flow and agitate. However, in this case, a structure such as a partition wall that can create a pressure difference within the bag is required. If a structure is provided inside the bag, the structure may hinder the cells from being discharged outside the bag, causing cells to remain in the bag and reducing the cell recovery rate.

[0007] One embodiment of the present invention suppresses a decrease in cell recovery rate during a stirring process of a cell suspension. [Means for solving the problem]

[0008] When the bag gripping clip according to the disclosed technology grips a bag that forms a storage space between a first film and a second film facing the first film, it applies a pressing force to the bag that partially seals the first film and the second film together, forming a partition that divides the storage space into multiple spaces that are connected to each other via communicating sections.

[0009] The partition may extend between a first side of the bag and a second side opposite the first side in a direction intersecting the first side and the second side, and a communication portion may be provided on the path along which the partition extends.

[0010] A communication portion may be provided between the partition wall and the first side or between the partition wall and the second side.

[0011] The bag gripping clip according to the disclosed technique is preferably detachable from the bag.

[0012] The partition wall may divide the storage space into a first space and a second space disposed vertically below the first space.

[0013] The partition wall may form a downward slope at the bottom of the first space, one end of which is connected to the lowest position of the bottom, and form a communication part that extends into the second space.

[0014] The bag gripping clip according to the disclosed technique may include a contact portion that includes an elastic body and that contacts the bag, and may further include a support member that supports the contact portion.

[0015] The pressing force that partially brings the first film and the second film into close contact with each other may be applied by magnetic force.

[0016] The bag gripping clip according to the disclosed technology may include a first abutment portion that abuts against the first film, a second abutment portion that abuts against a portion of the first film other than the portion that the first abutment portion abuts against, a third abutment portion that abuts against the second film and sandwiches the bag between the first abutment portion and the clip to form a portion of the partition separating the first space and the second space and a portion of the partition that forms the communicating portion, and a fourth abutment portion that abuts against the second film and sandwiches the bag between the second abutment portion and the clip to form another portion of the partition separating the first space and the second space and a portion of the partition that forms the communicating portion.

[0017] The bag gripping clip according to the disclosed technology may further include a first fixing portion that fixes the relative positions of the first abutment portion and the second abutment portion, and a second fixing portion that fixes the relative positions of the third abutment portion and the fourth abutment portion.

[0018] The bag gripping clip according to the disclosed technology may further include a first slide mechanism that changes the relative position of the first abutment portion and the second abutment portion by sliding at least one of the first abutment portion and the second abutment portion, and a second slide mechanism that changes the relative position of the first abutment portion and the second abutment portion by sliding at least one of the third abutment portion and the fourth abutment portion.

[0019] A container according to the disclosed technology includes the bag gripping clip described above and a bag gripped by the bag gripping clip.

[0020] The container according to the disclosed technology may further include a ventilation port connected to each of the plurality of spaces.

[0021] The stirring method of the disclosed technology is a stirring method for stirring a cell suspension using a container including a bag gripping clip that, when gripping a bag that forms a storage space between a first film and a second film facing the first film, applies a pressing force to the bag to partially seal the first film and the second film, forming a partition that divides the storage space into a first space and a second space that are connected to each other via a communicating part, and a bag gripped by the bag gripping clip, and the stirring method includes a first step of gripping the bag with the bag gripping clip, a second step of, after the first step, storing the cell suspension in the second space, and a third step of creating an air pressure difference between the first space and the second space and transferring the cell suspension stored in the second space to the first space via the communicating part.

[0022] The stirring method according to the disclosed technique may further include a fourth step of transferring the cell suspension contained in the first space to the second space via the communication part after the third step. Furthermore, the third step and the fourth step may be performed alternately and repeatedly.

[0023] The stirring method according to the disclosed technology may further include a fifth step of removing the bag gripping clip from the bag while the cell suspension is contained inside the bag, and a sixth step of discharging the cell suspension contained inside the bag to the outside of the bag after the fifth step.

[0024] The container according to the disclosed technology has an internal zip mechanism that can be opened and closed to partially seal a first film and a second film that face each other, and by closing the zip mechanism, a partition is formed that divides the storage space inside the container into multiple spaces that are connected to each other via communication sections. [Effects of the Invention]

[0025] According to one embodiment of the present invention, a decrease in the cell recovery rate can be suppressed during the stirring process of a cell suspension. [Brief explanation of the drawings]

[0026] [Figure 1] 1 is a perspective view schematically illustrating an example of the configuration of a bag gripping clip according to an embodiment of the disclosed technology. [Figure 2] FIG. 1 is a front view showing an example of a configuration of an agitation vessel according to an embodiment of the disclosed technology. [Figure 3] FIG. 3 is a cross-sectional view taken along line 3-3 in FIG. 2. [Figure 4] FIG. 1 is a diagram illustrating an example of an internal configuration of a stirring vessel according to an embodiment of the disclosed technology. [Figure 5A] 1A to 1C are diagrams illustrating an example of a stirring method according to an embodiment of the disclosed technology. [Figure 5B] 1A to 1C are diagrams illustrating an example of a stirring method according to an embodiment of the disclosed technology. [Figure 5C] 1A to 1C are diagrams illustrating an example of a stirring method according to an embodiment of the disclosed technology. [Figure 5D] 1A to 1C are diagrams illustrating an example of a stirring method according to an embodiment of the disclosed technology. [Figure 5E] 1A to 1C are diagrams illustrating an example of a stirring method according to an embodiment of the disclosed technology. [Figure 5F] 1A to 1C are diagrams illustrating an example of a stirring method according to an embodiment of the disclosed technology. [Figure 6A] FIG. 1 is a cross-sectional view showing an example of a configuration of a stirring vessel according to an embodiment of the disclosed technology. [Figure 6B] FIG. 1 is a cross-sectional view showing an example of a configuration of a stirring vessel according to an embodiment of the disclosed technology. [Figure 7] FIG. 10 is a cross-sectional view showing an example of the configuration of a stirring vessel according to another embodiment of the disclosed technology. [Figure 8] FIG. 10 is a diagram illustrating an example of an internal configuration of a stirring vessel according to another embodiment of the disclosed technology. [Figure 9A] 10A and 10B are diagrams illustrating an example of a stirring method according to another embodiment of the disclosed technology. [Figure 9B] 10A and 10B are diagrams illustrating an example of a stirring method according to another embodiment of the disclosed technology. [Figure 9C] 10A and 10B are diagrams illustrating an example of a stirring method according to another embodiment of the disclosed technology. [Figure 9D] 10A and 10B are diagrams illustrating an example of a stirring method according to another embodiment of the disclosed technology. [Figure 9E] 10A and 10B are diagrams illustrating an example of a stirring method according to another embodiment of the disclosed technology. [Figure 9F] 10A and 10B are diagrams illustrating an example of a stirring method according to another embodiment of the disclosed technology. [Figure 10] FIG. 10 is a diagram illustrating an example of an internal configuration of a stirring vessel according to another embodiment of the disclosed technology. [Figure 11A] FIG. 10 is a front view showing an example of the configuration of an agitation vessel according to another embodiment of the disclosed technology. [Figure 11B] FIG. 11B is a cross-sectional view taken along line 11B-11B in FIG. 11A. [Figure 12] FIG. 10 is a cross-sectional view showing an example of the configuration of a zip mechanism according to an embodiment of the disclosed technology. DETAILED DESCRIPTION OF THE INVENTION

[0027] Hereinafter, embodiments of the disclosed technology will be described with reference to the drawings. In each drawing, substantially the same or equivalent components or parts are designated by the same reference numerals.

[0028] [First embodiment] Fig. 1 is a perspective view schematically illustrating an example of the configuration of a bag gripping clip 1 according to an embodiment of the disclosed technology. Fig. 2 is a front view illustrating an example of the configuration of a stirring vessel 3 according to an embodiment of the disclosed technology, which includes the bag gripping clip 1 and a bag 2 gripped by the bag gripping clip 1. Fig. 3 is a cross-sectional view taken along line 3-3 in Fig. 2.

[0029] The bag 2 can be used, for example, for cell culture applications, and is constructed by bonding a first film F1 and a second film F2, each of which has flexibility and gas permeability, together by a method such as heat sealing (thermocompression bonding). The bag 2 may also be constructed by folding back a single film, and in this case, the opposing portions of the film are also referred to as the first film F1 and the second film F2, respectively.

[0030] The first film F1 and the second film F2 can be made of a suitable material, such as a plastic, polyethylene, or polypropylene. The bag 2 forms a storage space between the first film F1 and the second film F2 for storing cells together with a culture medium.

[0031] The bag gripping clip 1 is configured to include a first portion 10A arranged on the first film F1 side and a second portion 10B arranged on the second film F2 side, and grips the bag 2 by sandwiching the bag 2 between the first portion 10A and the second portion 10B. That is, the bag gripping clip 1 grips the bag 2 by applying a pressing force from the first film F1 side and the second film F2 side of the bag 2 to the other side. The bag gripping clip 1 is detachable from the bag 2.

[0032] The first part 10A of the bag gripping clip 1 has a linear first contact portion 11A that contacts the first film F1, a linear second contact portion 12A that contacts a portion of the first film F1 different from the portion that contacts the first contact portion 11A, and a support plate 13A that constitutes a support member that supports the first contact portion 11A and the second contact portion 12A. The first contact portion 11A and the second contact portion 12A are attached to a gripping surface Sa (see FIG. 3), which is the surface of the support plate 13A on which the bag 2 is placed.

[0033] Similarly, the second part 10B of the bag gripping clip 1 has a linear third contact portion 11B that contacts the second film F2, a fourth contact portion 12B that contacts a portion of the second film F2 different from the portion contacted by the third contact portion 11B, and a support plate 13B that constitutes a support member that supports the third contact portion 11B and the fourth contact portion 12B. The third contact portion 11B and the fourth contact portion 12B are attached to a gripping surface Sb (see FIG. 3) of the support plate 13B, which is the surface on the side where the bag 2 is placed.

[0034] 3, when the bag gripping clip 1 grips a bag 2, the first contact portion 11A and the third contact portion 11B overlap, and the second contact portion 12A and the fourth contact portion 12B overlap. That is, the bag 2 is sandwiched between the first contact portion 11A and the third contact portion 11B and between the second contact portion 12A and the fourth contact portion 12B. By sandwiching the bag 2 between the first contact portion 11A and the third contact portion 11B, the first film F1 and the second film F2 are brought into partial (linear) contact in accordance with the shapes of the first contact portion 11A and the third contact portion 11B, thereby forming a linear partition wall inside the bag 2. Similarly, by sandwiching the bag 2 between the second contact portion 12A and the fourth contact portion 12B, the first film F1 and the second film F2 are brought into partial (linear) contact according to the shapes of the second contact portion 12A and the fourth contact portion 12B, thereby forming a linear partition wall inside the bag 2. The first contact portion 11A, the second contact portion 12A, the third contact portion 11B, and the fourth contact portion 12B may each be made of an elastic material such as rubber. With this configuration, when the bag 2 is gripped with the bag gripping clip 1, the adhesion between the first film F1 and the second film F2 can be increased at the portions where the contact portions abut, and the function as a partition wall can be effectively exhibited.

[0035] 3, the first contact portion 11A and the second contact portion 12A are each attached to the support plate 13A via a spacer 14A, and the third contact portion 11B and the fourth contact portion 12B are each attached to the support plate 13B via a spacer 14B. By providing the spacers 14A and 14B, it is possible to ensure a sufficient space between the support plate 13A and the support plate 13B to place the bag 2 containing the cells together with the culture medium.

[0036] The relative positions of the first portion 10A and the second portion 10B of the bag gripping clip 1 are fixed by a plurality of screws 15 that penetrate the support plates 13A and 13B. Furthermore, the plurality of screws 15 apply a pressing force that causes the first film F1 and the second film F2 to be partially (linearly) in close contact with each other at the locations where the abutting portions of the bag 2 abut. That is, a pressing force that causes the first film F1 and the second film F2 to be partially (linearly) in close contact with each other is applied by the tensile elastic force exerted by the plurality of screws 15. Incidentally, a concave-convex structure for determining the relative positions of the first portion 10A and the second portion 10B may be provided on the support plates 13A and 13B near the screws 15.

[0037] 4 is a front view showing an example of the form of a partition wall 30 formed inside the bag 2 by gripping the bag 2 with the bag gripping clip 1, and is a diagram showing an example of the internal configuration of the stirring vessel 3. The partition wall 30 is formed in a portion of the bag 2 gripped by the bag gripping clip 1 where the first film F1 and the second film F2 are in close contact with each other.

[0038] The partition 30 divides the internal storage space of the bag 2 into a first space 31 and a second space 32. When a cell suspension in which cells are dispersed in a culture medium is stirred using the stirring vessel 3 including the bag gripping clip 1 and the bag 2, the second space 32 is located vertically below the first space 31. The partition 30 also forms a communication section 33 that connects the first space 31 and the second space 32. That is, the partition 30 has a portion extending in an intersecting direction between a side A1 that is an end of the rectangular bag 2 in a front view and a side A2 that is the other end of the bag 2 opposite to the side A1, and the communication section 33 is provided on an extension of the portion of the partition 30 that extends in the intersecting direction. More specifically, the partition wall 30 forms a downward slope at the bottom of the first space 31, and one end is connected to the lowest position of the sloped bottom of the first space 31, forming a communication part 33 that extends into the second space 32. The lowest position of the sloped bottom of the first space 31 is located in the horizontal center between sides A1 and A2 of the bag 2, and therefore the communication part 33 is also located in the horizontal center between sides A1 and A2.

[0039] The bag 2 has a ventilation port 41 connected to the first space 31 and a ventilation port 42 connected to the second space 32. Gas can be introduced into the interior of the bag 2 from the outside of the bag 2 via the ventilation ports 41 and 42. A liquid feed port 43 is provided at the bottom of the second space 32, and the cell suspension can be discharged from the interior of the bag 2 or injected into the interior of the bag 2 via the liquid feed port 43. The bag 2 may have multiple liquid feed ports, including a liquid feed port dedicated to injection for injecting the cell suspension into the interior of the bag 2 and a liquid feed port dedicated to discharge for discharging the cell suspension from the interior of the bag 2.

[0040] 5A to 5F, a stirring method according to an embodiment of the disclosed technology for stirring a cell suspension using a stirring vessel 3 will be described below. Note that in FIGS. 5B to 5E, a partition wall 30 formed inside the bag 2 by gripping the bag 2 with the bag gripping clip 1 is shown, and the bag gripping clip 1 is not shown.

[0041] First, a bag 2 containing no cell suspension is prepared (FIG. 5A). Next, the bag 2 is gripped with the bag gripping clip 1 (FIG. 5B). Specifically, the bag 2 is clamped between the first portion 10A and the second portion 10B of the bag gripping clip 1, and the screw 15 is tightened. This applies a pressing force that causes the first film F1 and the second film F2 of the bag 2 to be partially (linearly) adhered to each other by the first abutting portion 11A, the second abutting portion 12A, the third abutting portion 11B, and the fourth abutting portion 12B. By gripping the bag 2 with the bag gripping clip 1, a partition wall 30 is formed inside the bag 2. The partition wall 30 divides the storage space inside the bag 2 into a first space 31 and a second space 32 that are connected to each other via a communication portion 33. By gripping the bag 2 with the bag gripping clip 1, a stirring vessel 3 is formed.

[0042] Next, the orientation of the agitator 3 is fixed so that the second space 32 of the agitator 3 is positioned vertically below the first space 31, and the cell suspension 100 is injected into the bag 2 through the liquid supply port 43 (FIG. 5C). The cell suspension 100 is contained in the second space 32 of the agitator 3. Note that the amount of cell suspension 100 injected into the bag 2 is preferably smaller than the capacity of the second space 32 so that the second space 32 is not entirely filled with the cell suspension 100.

[0043] Next, while the ventilation port 41 connected to the first space 31 is left open, gas is introduced into the second space 32 through the ventilation port 42 connected to the second space 32. This causes the air pressure inside the second space 32 to become higher than the air pressure inside the first space 31. As a result, the liquid surface of the cell suspension 100 contained in the second space 32 is pressurized, and the cell suspension 100 is transferred to the first space 31 via the communication part 33. By transferring the cell suspension 100 to the first space 31 via the communication part 33, the cell suspension 100 is stirred (FIG. 5D).

[0044] The gas introduced into the bag 2 is preferably sterile to prevent contamination of the cells. A gas harmless to cells, such as oxygen or air, is preferably used as the gas introduced into the bag 2. While the above description illustrates a case in which the cell suspension 100 is transferred from the second space 32 to the first space 31 by pressurizing the interior of the second space 32, the present invention is not limited to this. Specifically, the cell suspension 100 may be transferred from the second space 32 to the first space 31 by reducing the pressure inside the first space 31, for example, by sucking the gas inside the first space 31 through the ventilation port 41.

[0045] Next, the ventilation port 42 connected to the second space 32 is opened. As a result, the cell suspension 100 contained in the first space 31 is transferred by gravity to the second space 32 via the communication part 33 (FIG. 5E). The partition 30 forms a downward slope at the bottom of the first space 31, and the communication part 33 is formed at the lowest position of the sloped bottom. This allows the cell suspension 100 contained in the first space 31 to be guided to the communication part 33 by gravity. The cell suspension 100 is transferred to the second space 32 via the communication part 33, and the cell suspension 100 is agitated. Note that the transfer of the cell suspension 100 from the first space 31 to the second space 32 may be promoted by introducing gas into the first space 31 via the ventilation port 41 connected to the first space 31.

[0046] Thereafter, if necessary, the transfer of the cell suspension 100 from the second space 32 to the first space 31 and the transfer of the cell suspension 100 from the first space 31 to the second space 32 may be repeated multiple times. The transfer of the cell suspension 100 is preferably repeated until the cells are dispersed approximately uniformly in the medium.

[0047] Next, the bag gripping clip 1 is removed from the bag 2. This removes the partition wall 30 separating the first space 31 and the second space 32. Thereafter, the cell suspension 100 contained inside the bag 2 is discharged to the outside of the bag 2 through the liquid delivery port 43 (FIG. 5F). Note that after the bag gripping clip 1 is removed from the bag 2, cell culture may be performed in the bag 2.

[0048] As described above, when the bag gripping clip 1 according to the embodiment of the disclosed technology grips the bag 2, it applies a pressure that partially (linearly) brings the first film F1 and the second film F2 of the bag 2 into close contact, forming a partition wall 30 that divides the storage space of the bag 2 into a first space 31 and a second space 32 that are connected to each other via a communication section 33. Furthermore, a stirring method according to the embodiment of the disclosed technology grips the bag 2 with the bag gripping clip 1, introduces a cell suspension into the bag 2, and uses the pressure difference between the first space 31 and the second space 32 or gravity to cause the cell suspension to flow between the first space 31 and the second space 32, thereby stirring the cell suspension. This method of stirring the cell suspension by using a pressure difference or gravity to cause the cell suspension to flow can reduce damage to cells compared to a method using a stirring blade. To more reliably suppress cell damage, it is preferable to strictly control the pressure applied to the liquid surface using a pump or the like.

[0049] Furthermore, the stirring method according to the embodiment of the disclosed technology can uniformly disperse cells in the culture medium, similar to the method using a stirring blade. After the stirring process, the bag gripping clip 1 is removed from the bag 2, thereby removing the partition wall 30 inside the bag 2. This eliminates any structures that may hinder the cell suspension from being discharged outside the bag 2, thereby preventing cells from remaining inside the bag 2. This prevents a decrease in the cell recovery rate.

[0050] The stirring method according to this embodiment can also be used for the purpose of mixing cells and medium at predetermined times during the culture period, for example, in static culture in which cells are left stationary in a suspended state in the medium. This improves the dispersion state of cells dispersed in the medium, and creates a favorable culture environment in which nutrients and gases are distributed evenly to all cells.

[0051] Here, when attaching the bag gripping clip 1 to the bag 2, the following operation may be performed to promote the formation of a space through which the cell suspension flows in the communication part 33. Note that Figures 6A and 6B referred to in the following description are cross-sectional views showing an example of the configuration of the stirring vessel 3, and show a cross section corresponding to the cross section shown in Figure 3.

[0052] 6A, with the bag 2 sandwiched between the first portion 10A and the second portion 10B of the bag gripping clip 1, the screws 15 are temporarily tightened. The bag 2 is sandwiched between the first abutting portion 11A and the third abutting portion 11B and between the second abutting portion 12A and the fourth abutting portion 12B.

[0053] Next, with the bag 2 sandwiched between the first abutment portion 11A and the third abutment portion 11B, and between the second abutment portion 12A and the fourth abutment portion 12B, the first abutment portion 11A and the second abutment portion 12A are slid in a direction in which they approach each other, and the third abutment portion 11B and the fourth abutment portion 12B are slid in a direction in which they approach each other.

[0054] 6B, in the region of the bag 2 between the portion sandwiched between the first contact portion 11A and the third contact portion 11B and the portion sandwiched between the second contact portion 12A and the fourth contact portion 12B, the first film F1 and the second film F2 are each deflected so as to bulge outward from the bag 2. As a result, a space B through which the cell suspension flows is formed in the communication portion 33. Alternatively, one of the first contact portion 11A and the second contact portion 12A may be slid toward the other, and one of the third contact portion 11B and the fourth contact portion 12B, which faces the slid first contact portion 11A or the second contact portion 12A, may be slid toward the other.

[0055] After forming the space B in the communication portion 33, the positions of the first contact portion 11A and the second contact portion 12A are fixed with the screws 16. This fixes the relative positions of the first contact portion 11A and the second contact portion 12A. Similarly, the positions of the third contact portion 11B and the fourth contact portion 12B are fixed with the screws 16. This fixes the relative positions of the third contact portion 11B and the fourth contact portion 12B. Thereafter, the screws 15 are fully tightened.

[0056] When the space B is formed in the communication portion 33 in the above manner, the bag gripping clip 1 includes a first slide mechanism that changes the relative position between the first contact portion 11A and the second contact portion 12A, a second slide mechanism that changes the relative position between the third contact portion 11B and the fourth contact portion 12B, a first fixing portion that fixes the relative position between the first contact portion 11A and the second contact portion 12A, and a second fixing portion that fixes the relative position between the third contact portion 11B and the fourth contact portion 12B. Note that the first and second slide mechanisms may be configured to slide the spacers 14A and 14B, to which the respective contact portions are joined, integrally with the respective contact portions. The first and second fixing portions may be configured to include screws 16 that fix the spacers 14A and 14B, to which the contact portions are joined, to the support plates 13A and 13B, respectively.

[0057] Furthermore, when space B is formed in communication portion 33 in the manner described above, it is necessary to either secure a volume of gas or liquid equivalent to the volume of space B inside bag 2 in advance, or to introduce it from the outside of bag 2 when space B is formed. For example, when space B is formed, one of ventilation ports 41 and 42 may be left open, and outside air may be introduced into bag 2 through the open ventilation port. The gas introduced into bag 2 is preferably in a sterile state to prevent contamination. Furthermore, when bag 2 is washed in advance, the cleaning liquid used for washing may be left inside bag 2.

[0058] [Second embodiment] 7 is a cross-sectional view showing an example of the configuration of an agitation vessel 3A including a bag gripping clip 1A according to a second embodiment of the disclosed technology and a bag 2 gripped by the bag gripping clip 1 A. Fig. 7 shows a cross section corresponding to the cross section shown in Fig. 3.

[0059] The bag gripping clip 1 according to the first embodiment described above applies a pressing force that brings the first film F1 and the second film F2 into partial (linear) contact with each other by using a tensile elastic force exerted by a plurality of screws 15. In contrast, the bag gripping clip 1A according to the second embodiment applies a pressing force that brings the first film F1 and the second film F2 into partial (linear) contact with each other by using a magnetic force exerted by a magnet 40A attached to a support plate 13A and a magnet 40B attached to a support plate 13B. The magnets 40A and 40B may be permanent magnets or electromagnets.

[0060] The bag gripping clip 1A according to the second embodiment reduces the workload required to attach the bag gripping clip 1A to the bag 2 compared to the bag gripping clip 1 according to the first embodiment. Furthermore, by configuring the magnets 40A and 40B as electromagnets, it becomes possible to electrically control the generation of magnetic force. For example, when removing the bag gripping clip 1A from the bag 2, stopping the generation of magnetic force makes it easier to remove the bag gripping clip 1A. The first contact portion 11A, the second contact portion 12A, the third contact portion 11B, and the fourth contact portion 12B may be configured with magnets. Furthermore, each of the spacers 14A and 14B may be configured with a magnet.

[0061] [Third embodiment] 8 is a plan view showing another form of the partition wall 30 formed inside the bag 2 by gripping the bag 2 with a bag gripping clip (not shown), and is a diagram showing an example of the internal configuration of a stirring vessel 3B according to a third embodiment of the disclosed technology. The bag gripping clip according to this embodiment (not shown) includes a contact portion that contacts the bag 2 in the portion where the partition wall 30 shown in FIG. 8 is formed, and a support plate that supports the contact portion. The partition wall 30 is formed in the portion of the bag 2 gripped by the bag gripping clip where the first film F1 and the second film F2 are in close contact with each other.

[0062] The partition 30 divides the storage space inside the bag 2 into a first space 31 and a second space 32. When a cell suspension is stirred using the stirring vessel 3B, the first space 31 and the second space 32 are arranged side by side in the horizontal direction. The partition 30 also forms a communication section 33 that connects the first space 31 and the second space 32. That is, in the stirring vessel 3B, the partition 30 has a portion extending in a transverse direction that intersects with the sides A3 and A4 between a side A3 that is an end of the rectangular bag 2 when viewed from the front and a side A4 that is the other end of the bag 2 opposite the side A3. The communication section 33 is provided on an extension of the portion of the partition 30 extending in the transverse direction. More specifically, the partition 30 has a portion that extends linearly from the side A3 to the side A4 and a portion that extends linearly from the side A4 to the side A3. The gap between the two portions forms the communication section 33.

[0063] The bag 2 has a ventilation port 41 connected to the first space 31 and a ventilation port 42 connected to the second space 32. Gas can be introduced into the interior of the bag 2 from the outside of the bag 2 via the ventilation ports 41 and 42. In addition, a liquid supply port 43 is provided at the bottom of the bag 2, and a cell suspension can be discharged from the interior of the bag 2 or injected into the interior of the bag 2 via the liquid supply port 43.

[0064] Hereinafter, a stirring method according to an embodiment of the disclosed technology, in which a cell suspension is stirred using the stirring vessel 3B, will be described with reference to FIGS. 9A to 9F.

[0065] First, a bag 2 is prepared without containing a cell suspension (FIG. 9A). Next, the bag 2 is gripped with a bag gripping clip (FIG. 9B). This forms a partition wall 30 inside the bag 2, and the internal storage space of the bag 2 is divided by the partition wall 30 into a first space 31 and a second space 32 that communicate with each other via a communication portion 33. By gripping the bag 2 with the bag gripping clip, a stirring vessel 3B is formed.

[0066] Next, the orientation of the agitator 3B is fixed so that the first space 31 and the second space 32 of the agitator 3B are arranged horizontally side by side, and the cell suspension 100 is injected into the agitator 3B through the liquid supply port 43 (FIG. 9C). The cell suspension 100 is evenly distributed into the first space 31 and the second space 32.

[0067] Next, while the ventilation port 42 connected to the second space 32 is left open, gas is introduced into the first space 31 through the ventilation port 41 connected to the first space 31. This causes the air pressure inside the first space 31 to become higher than the air pressure inside the second space 32. As a result, the liquid surface of the cell suspension 100 contained in the first space 31 is pressurized, and a portion of the liquid is transferred to the second space 32 via the communication part 33. By transferring the cell suspension 100 to the second space 32 via the communication part 33, the cell suspension 100 is stirred (FIG. 9D).

[0068] Next, while the ventilation port 41 connected to the first space 31 is left open, gas is introduced into the second space 32 through the ventilation port 42 connected to the second space 32. This causes the air pressure inside the second space 32 to become higher than the air pressure inside the first space 31. As a result, the liquid surface of the cell suspension 100 contained in the second space 32 is pressurized, and a portion of the liquid is transferred to the first space 31 via the communication part 33. By transferring the cell suspension 100 to the first space 31 via the communication part 33, the cell suspension 100 is stirred (FIG. 9E).

[0069] Thereafter, if necessary, the transfer of the cell suspension 100 from the first space 31 to the second space 32 and the transfer of the cell suspension 100 from the second space 32 to the first space 31 may be repeated multiple times. The transfer of the cell suspension 100 is preferably repeated until the cells are dispersed approximately uniformly in the medium.

[0070] Next, the bag gripping clip is removed from the bag 2. This removes the partition wall 30 separating the first space 31 and the second space 32. Thereafter, the cell suspension 100 contained inside the bag 2 is discharged to the outside of the bag 2 through the liquid delivery port 43. Note that after the bag gripping clip is removed from the bag 2, cell culture may be performed in the bag 2.

[0071] As described above, even in the agitation vessel 3B having the partition wall 30 in the form shown in Fig. 8, it is possible to agitate the cell suspension in the same manner as the agitation method using the agitation vessel 3 according to the first embodiment. Therefore, in the agitation process of the cell suspension, damage to the cells can be reduced and a decrease in the cell recovery rate can be suppressed.

[0072] [Fourth embodiment] 10 is a plan view showing another form of partition wall 30 formed inside bag 2 by gripping bag 2 with a bag gripping clip (not shown), and is a diagram showing an example of the internal configuration of a stirring vessel 3C according to a fourth embodiment of the disclosed technology. The bag gripping clip according to this embodiment (not shown) includes a contact portion that contacts bag 2 in the portion where partition wall 30 shown in FIG. 10 is formed, and a support plate that supports the contact portion. Partition wall 30 is formed in the portion of bag 2 gripped by bag gripping clip 1 where first film F1 and second film F2 are in close contact with each other.

[0073] The partition 30 divides the storage space inside the bag 2 into a first space 31 and a second space 32. When a cell suspension is stirred using the stirring vessel 3C, the first space 31 and the second space 32 are arranged side by side in the horizontal direction. The partition 30 also forms a communication section 33 that connects the first space 31 and the second space 32. That is, in the stirring vessel 3C, the partition 30 has a portion extending in a cross direction intersecting the sides A3 and A4 between a side A3 that is an end of the rectangular bag 2 in a front view and a side A4 that is the other end of the bag 2 opposite the side A3, and the communication section 33 is provided on an extension of the portion of the partition 30 extending in the cross direction. More specifically, the partition 30 has a portion that extends linearly from the side A3 to the side A4, and the communication section 33 is provided between the partition 30 and the side A4. The partition wall 30 may have a portion that extends linearly from the side A4 toward the side A3, and a communication portion 33 may be provided between the partition wall 30 and the side A3.

[0074] The bag 2 has a ventilation port 41 connected to the first space 31 and a ventilation port 42 connected to the second space 32. Gas can be introduced into the interior of the bag 2 from the outside of the bag 2 via the ventilation ports 41 and 42. In addition, a liquid supply port 43 is provided at the bottom of the bag 2, and a cell suspension can be discharged from the interior of the bag 2 or injected into the interior of the bag 2 via the liquid supply port 43.

[0075] The method for stirring the cell suspension using the stirring vessel 3C is the same as that for using the stirring vessel 3B according to the third embodiment described above, and therefore a description thereof will be omitted.

[0076] [Fifth embodiment] Figure 11A is a front view showing an example of the configuration of a stirring vessel 3D according to a fifth embodiment of the disclosed technology, Figure 11B is a cross-sectional view taken along line 11B-11B in Figure 11A, and Figure 12 is an enlarged view of the area surrounded by the dashed line in Figure 11B, and is a cross-sectional view showing an example of the configuration of a zip mechanism 50.

[0077] As shown in FIG. 11A, the agitator 3D has a partition wall 30 therein, similar in configuration to the agitator 3 according to the first embodiment (see FIG. 4). In the agitator 3 according to the first embodiment, the partition wall 30 is formed by gripping the bag 2 with the bag gripping clip 1. In contrast, the agitator 3D according to this embodiment has an internal zip mechanism 50 that can be opened and closed to linearly bring the opposing first film F1 and second film F2 into close contact, and the partition wall 30 is formed by closing the zip mechanism 50. The partition wall 30 divides the storage space inside the agitator 3D into a first space 31 and a second space 32. The partition wall 30 also forms a communication section 33 that connects the first space 31 and the second space 32.

[0078] 12, the zip mechanism 50 includes a protrusion 51 formed on the inner surface of the first film F1 and a recess 52 disposed opposite the protrusion 51 on the inner surface of the second film F2. By fitting the protrusion 51 into the recess 52, the zip mechanism 50 is closed, and the first film F1 and the second film F2 come into close contact at the formation site of the zip mechanism 50, forming a partition wall 30 inside the agitator vessel 3D. Furthermore, by releasing the fitting between the protrusion 51 and the recess 52, the zip mechanism 30 is opened, and the partition wall 30 is removed from inside the agitator vessel 3D.

[0079] The agitator 3D is equipped with a ventilation port 41 connected to the first space 31 and a ventilation port 42 connected to the second space 32. Gas can be introduced into the interior of the agitator 3D from the outside of the agitator 3D via the ventilation ports 41 and 42. In addition, a liquid feed port 43 is provided at the bottom of the second space 32, and via the liquid feed port 43, a cell suspension can be discharged from the interior of the agitator 3D or injected into the interior of the agitator 3D.

[0080] The method for stirring the cell suspension using the agitator 3D is the same as when using the agitator 3 according to the first embodiment described above. With the agitator 3D, after the stirring process, the zip mechanism 50 is opened to remove the partition 30 inside the agitator 3D, and when the cell suspension is discharged to the outside of the agitator 3D, there is no structure that would hinder discharge, so it is possible to prevent cells from remaining inside the agitator 3D. This makes it possible to prevent a decrease in the cell recovery rate.

[0081] In the stirring vessel 3D, the shape of the partition wall 30 may be the same as the shape of the partition wall 30 of the stirring vessel 3B shown in FIG. 8, or may be the same as the shape of the partition wall 30 of the stirring vessel 3C shown in FIG.

[0082] This application claims priority from Japanese Patent Application No. 2017-184952, filed September 26, 2017, the entire contents of which are incorporated herein by reference. In addition, all publications, patent applications, and technical standards described herein are incorporated herein by reference to the same extent as if each individual publication, patent application, and technical standard was specifically and individually indicated to be incorporated by reference.

Claims

[Claim 1] A bag gripping clip that, when gripping a bag that forms a storage space between a first film and a second film facing the first film, applies a pressing force to the bag that partially brings the first film and the second film into close contact with each other, thereby forming a partition that divides the storage space into a first space and a second space that are communicated with each other via a communication part, It is detachable from the bag, the partition wall has a portion extending in a cross direction intersecting the first side and the second side of the bag between the first side and the second side opposite to the first side, The communication portion is provided between a portion of the partition wall extending from the first side toward the second side and a portion of the partition wall extending from the second side toward the first side. Clip for holding bags.

Images