Stirring device, stirring system, and successive cultivation method

Abstract

The present invention comprises: an external structure that has a hollow shape; an internal structure that is disposed inside the external structure and that defines an annular flow path which accommodates a culture solution and air between the internal structure and an inner peripheral surface the external structure; and a driving unit that applies an external force in the circumferential direction of the annular flow path to the culture solution in the annular flow path, wherein the external structure is provided with an air supply unit that supplies air downward from above, toward the liquid surface of the culture solution accommodated in the annular flow path.

Description

Stirring device, stirring system, and subculture method

[0003]

[0001] The present disclosure relates to a stirring device, a stirring system, and a subculture method. This application claims priority to Japanese Patent Application No. 2025-004353 filed in Japan on January 10, 2025, the content of which is incorporated herein by reference.

[0002] Patent Document 1 discloses a stirring device that fills a culture solution in an annular flow path between an outer cylinder and an inner cylinder and blows gas from below into the annular flow path. According to this stirring device, the culture solution is swirled in the annular flow path by blowing gas into the culture solution, thereby promoting the stirring of the culture solution.

[0003] Japanese Patent No. 4079877

[0004] However, in a stirring device that stirs by swirling a culture solution as in Patent Document 1, when performing suspension culture of animal cells, sufficient contact between the culture solution (cell suspension) and the gas in the gas phase above the liquid surface of the culture solution may not be obtained. Therefore, there is a possibility that the culture solution cannot be maintained at a desired hydrogen ion exponent (pH).

[0005] The present disclosure has been made in view of the above circumstances, and an object thereof is to provide a stirring device, a stirring system, and a subculture method capable of promoting gas-liquid contact between a liquid phase and a gas phase.

[0006] In order to solve the above problems, a stirring device according to the present disclosure includes a hollow external structure, an internal structure disposed within the external structure and partitioning an annular flow path for accommodating a culture solution and air between the internal peripheral surface of the external structure, and a driving unit that applies an external force in the circumferential direction of the annular flow path to the culture solution in the annular flow path. The external structure includes an air supply unit that supplies air from above downward toward the liquid surface of the culture solution accommodated in the annular flow path.

[0007] The subculture system according to this disclosure is a subculture system using the above-described stirring device, comprising a plurality of stirring devices: a first stirring device for performing subculture and a second stirring device for measuring the culture medium, wherein the second stirring device is equipped with a measuring liquid supply unit capable of supplying a measuring liquid for measurement to the annular flow path, and a plurality of liquid discharge units provided in one of the first stirring devices include a first liquid discharge unit that communicates with the liquid supply unit of an adjacent first stirring device and a second liquid discharge unit that communicates with the liquid supply unit of the second stirring device.

[0008] The subculture method according to this disclosure is a subculture method using the subculture system described above, comprising: a subculture step in which subculture is performed sequentially using a plurality of first stirring devices connected via the first liquid discharge unit; and a measurement step in parallel with the subculture step in which a portion of the culture medium of each generation cultured by the plurality of first stirring devices is supplied via the second liquid discharge unit to the annular channel of the second stirring device corresponding to each generation, and the culture medium is measured.

[0009] According to this disclosure, it is possible to promote gas-liquid contact between the liquid phase and the gas phase.

[0010] This is a cross-sectional view showing the schematic configuration of a stirring device in the first embodiment of the present disclosure. This is a cross-sectional view of the stirring device in Figure 1, viewed from the right. This is a cross-sectional view showing the state in which culture is being performed using the stirring device. This is a cross-sectional view showing the state in which culture is being removed from the stirring device. This is a cross-sectional view of a stirring device in the second embodiment of the present disclosure. This is a cross-sectional view showing the schematic configuration of a stirring device in the third embodiment of the present disclosure. This is a cross-sectional view of a stirring device in the fourth embodiment of the present disclosure. This is a cross-sectional view of the external structure body in a modified example of the fourth embodiment of the present disclosure. This is a cross-sectional view of the external structure body in Figure 8, viewed from the right. This is a cross-sectional view showing the schematic configuration of a subculture system in the fifth embodiment of the present disclosure.

[0011] <First Embodiment> The first embodiment of the present disclosure will now be described in detail with reference to Figures 1 to 4. Figure 1 is a cross-sectional view showing the schematic configuration of a stirring device in the first embodiment of the present disclosure. Figure 2 is a cross-sectional view of the stirring device of Figure 1 viewed from the right. Figure 3 is a cross-sectional view showing the state in which culture is being carried out using the stirring device. Figure 4 is a cross-sectional view showing the state in which the culture medium is being removed from the stirring device. <Stirring Device> The stirring device 1 of the first embodiment is a device used for cell culture. As shown in Figures 1 to 4, the stirring device 1 comprises an external structure 10, an internal structure 40, and a drive unit 50.

[0012] <External Structure> The external structure 10 is a component that forms the outer shape of the stirring device 1 and has a hollow interior. The external structure 10 has an external structure body 11 and a lid 12.

[0013] <External Structure Body> The external structure body 11 is a container capable of holding liquid inside and has an upper opening that opens upward. The external structure body 11 has a curved surface 20 and a pair of end plates 30. The curved surface 20 is formed in an arc shape centered on an axis O1 that extends in a first direction D1 which is horizontal. The outer circumferential surface of the curved surface 20 is an arc outer circumferential surface 21 that forms an arc shape centered on the axis O1. The inner circumferential surface of the curved surface 20 is an arc inner circumferential surface 22 that forms an arc shape centered on the axis O1. The curved surface 20 is formed in a plate shape with a constant thickness, with the radial direction being the thickness direction, centered on the axis O1.

[0014] The curved surface portion 20 has a first end 20ta and a second end 20tb in the circumferential direction centered on the axis O1. The first end 20ta and the second end 20tb form the upper end of the curved surface portion 20, which is the uppermost part of the curved surface portion 20. The first end 20ta and the second end 20tb are spaced apart in the second direction D2, which is a horizontal direction perpendicular to the first direction D1.

[0015] The pair of end plates 30 are positioned to sandwich the curved portion 20 from both sides in the first direction D1. In other words, the pair of end plates 30 close both ends of the curved portion 20 in the first direction D1. The pair of end plates 30 and the curved portion 20 form a housing space within the main body 11 of the external structure. The upper edges of the pair of end plates 30 extend in the second direction D2, and together with the first end 20ta and the second end 20tb, they form an upper opening 13 that opens upward.

[0016] <Lid> The lid 12 is formed to open and close the upper opening 13 of the external structure body 11. The structure for opening and closing the lid 12 can be selected as appropriate, and examples include a screw-in type or a hinge type. The lid 12 of this embodiment is equipped with an air supply unit 14 and an air discharge unit 15, which will be described later.

[0017] <Internal Structure> The internal structure 40 is located inside the external structure 10. In this embodiment, the internal structure 40 is a rotating body 41 that can rotate around an eccentric axis O2. The rotating body 41 is cylindrical in shape and extends around the eccentric axis O2. The rotating body 41 extends in a first direction D1 within the external structure 10. The rotating body 41 has a cylindrical outer surface 43 centered on the eccentric axis O2. The outer diameter of the cylindrical outer surface 43 is smaller than the inner diameter of the curved surface portion 20 of the external structure 10.

[0018] The rotating body 41 is positioned within the external structure 10 with an offset to one side of the second direction D2 (the right side in Figures 1 and 3). That is, the eccentric axis O2 is positioned eccentrically so as to be shifted to one side of the second direction D2 relative to the central axis O1.

[0019] Because the rotating body 41 is positioned biased towards one side in the second direction D2, the portion of the internal structure 40 in the annular flow path on that side in the second direction D2 has a reduced flow path cross-sectional area. This region where the flow path cross-sectional area is reduced is called the reduced region R1 of the annular flow path. The reduced region R1 is the portion of the range of the rotating body 41 in the vertical direction D3 of the annular flow path that is on the side of the eccentric axis O2 in the second direction D2.

[0020] Because the rotating body 41 is positioned biased towards one side in the second direction D2, the portion of the internal structure 40 in the annular flow channel on the other side of the second direction D2 (the left side in Figures 1 and 3) has an enlarged flow channel cross-sectional area. This region where the flow channel cross-sectional area is enlarged is called the enlarged region R2 of the annular flow channel. The enlarged region R2 is the portion of the range of the rotating body 41 in the vertical direction D3 of the annular flow channel that is on one side of the eccentric axis O2 in the second direction D2.

[0021] <Drive Unit> The drive unit 50 applies an external force to the culture medium C in the annular channel in the circumferential direction of the annular channel. The drive unit 50 is an electric motor 51 that rotates the rotating body 41 around the eccentric axis O2. The drive unit 50 may also be configured to use a power source other than the electric motor 51. The electric motor 51 has an electric motor body 51a and an output shaft 51b. Here, an example of a power source other than the electric motor 51 is a configuration in which the rotating body 41 is rotated non-contact from outside the external structure 10 using magnetic force.

[0022] The motor body 51a is located outside the external structure 10. When the motor body 51a is driven, the output shaft 51b is rotationally driven. The output shaft 51b is a shaft that extends in the first direction D1 and passes through the end plate 30 and is connected to the rotating body 41. As a result, the rotating body 41 is rotationally driven around the eccentric axis O2 as the output shaft 51b rotates. Note that if the drive unit 50 rotates the rotating body 41 without contact as described above, the configuration in which the output shaft 51b passes through the end plate 30 can be omitted.

[0023] Here, the rotating body 41 is rotated in the stirring direction by the electric motor 51. That is, as shown in Figure 3, the cylindrical outer surface 43 of the rotating body 41 rotates in the stirring direction (clockwise in Figures 1 and 3), progressing sequentially from the expanding region R2, the top 44a of the rotating body 41, the contracting region R1, the bottom 44b of the rotating body 41, the expanding region R2, the top 44a, and so on. As a result, an external force is applied to the culture medium C in the annular channel in the circumferential direction of the annular channel, moving from the expanding region R2 to the contracting region R1 above the internal structure.

[0024] <Culture medium and air> Culture medium C and air are supplied into the annular channel. Furthermore, the culture medium C and air are discharged from within the annular channel. For this reason, the external structure 10 has an air supply section 14, an air discharge section 15, a liquid discharge section 23, and a gas discharge section 24.

[0025] <Air Supply Unit> The air supply unit 14 is formed to supply air to the annular channel. The air supply unit 14 supplies air from above downward toward the liquid surface LS of the culture medium C contained in the annular channel. The air supply unit 14 in this embodiment is a hole that penetrates the external structure 10 inward and outward, and is formed in the lid 12. The air supply unit 14 is connected to an air source (not shown) located outside the external structure 10. The air supplied from the air source contains, for example, more carbon dioxide than in the atmosphere. The air supplied from the air source may be humidified to prevent evaporation of the culture medium C.

[0026] The air supply unit 14 supplies air toward the liquid surface L2 of the expanded region R2. In this embodiment, the air supply unit 14 supplies air diagonally from above toward below so that it forms a counterflow Cf (see Figure 3) opposite to the direction of flow of the culture medium C at the liquid surface LS (liquid surface L2) of the culture medium C. In this embodiment, the air supply unit 14 is located on one side of the second direction D2 (right side in Figures 1 and 3). In other words, the air supply unit 14 in this embodiment is located vertically above the liquid surface L1 of the reduced region R1. The air supplied from the air supply unit 14 into the external structure 10 flows from above on one side of the second direction D2 where the air supply unit 14 is located toward the liquid surface L2 diagonally downward on the other side of the second direction D2 (left side in Figures 1 and 3), collides with the liquid surface L2, and then becomes a counterflow Cf that flows along the liquid surface L2.

[0027] <Air Discharge Section> The air discharge section 15 is capable of exhausting air from the annular flow path. The air discharge section 15 is a hole that penetrates the external structure 10 from the inside to the outside. In this embodiment, the air discharge section 15 is provided on the upper part of the external structure 10. The air discharge section 15 is provided so as to penetrate the annular flow path and the outside of the external structure 10 in the vertical direction D3. The air discharge section 15 is positioned above the rotating body 41, and in this embodiment, it is positioned at the very top of the lid 12 that forms the upper part of the external structure 10. The air discharge section 15 is formed at an upper position opposite the liquid surface L2 of the enlarged region R2. In other words, the air supply section 14 described above is formed in the lid 12, and the air discharge section 15 may be open to the atmosphere outside the external structure 10, or it may be connected to a suction device provided outside the external structure 10.

[0028] Here, when the suction device connected to the air discharge section 15 sucks in gaseous air above the liquid level LS in the annular flow path, negative pressure is generated at the air discharge section 15. This negative pressure slightly raises the liquid level L2 in the expanded region R2 facing the air discharge section 15. As a result, the height difference between the liquid level L2 in the expanded region R2 and the liquid level L1 in the contracted region R1 can be increased, allowing for smoother overflow of the internal structure 40, which will be described later.

[0029] The air discharge section 15 or the connection point (not shown) to the air discharge section 15 is provided with a mechanism (not shown), such as an on / off valve, to stop the outflow of air from the air discharge section 15. For example, by stopping the outflow of air from the air discharge section 15 with the mechanism (not shown) and continuing the supply of air by the air supply section 14, it is possible to increase the pressure of the gas phase above the liquid level LS in the annular flow path.

[0030] Furthermore, the air discharge section 15 is equipped with a defoaming section 16 that breaks down bubbles generated in the culture medium C by coming into contact with them. The defoaming section 16 can be formed from wire mesh, perforated metal, or the like. The defoaming section 16 is positioned between the air discharge section 15 and the liquid surface L2, and is positioned to cover the holes of the air discharge section 15 from below.

[0031] <Liquid Discharge Section> The liquid discharge section 23 is capable of connecting the annular channel with the outside of the external structure 10. The liquid discharge section 23 forms a channel that can discharge the culture medium C contained in the annular channel to the outside of the external structure 10. In this embodiment, the liquid discharge section 23 is formed in a tubular shape that extends diagonally downward from the lower part of the curved surface section 20 and opens diagonally downward. The liquid discharge section 23 has a first opening / closing section 25 that opens and closes the channel within the liquid discharge section 23. As shown in Figure 4, this first opening / closing section 25 allows, for example, air A1 to be supplied to the gas phase above the liquid level LS to increase the pressure of the gas phase, and by opening the first opening / closing section 25 from a closed state, the culture medium C in the annular channel can be easily pushed out through the liquid discharge section 23 to a container 60 or the like placed outside the external structure 10. Examples of purposes for removing the culture medium C to a container 60 or the like include subculturing in subculture, cell count measurement, and glucose concentration measurement. Furthermore, when performing fed-batch culture, antibody measurement of cell suspension can be exemplified.

[0032] <Gas Release Section> The gas release section 24 is formed to release gas into the culture medium C in the annular channel. The gas release section 24 has a hole that penetrates the external structure body 11 both internally and externally. The gas release section 24 is provided to connect the inside of the curved section 20 with the outside of the curved section 20. The gas release section 24 is connected to a gas supply source (not shown) located outside the external structure 10. Culture oxygen from the gas supply source is supplied into the annular channel via the gas release section 24.

[0033] <Operation of the stirring device> As shown in Figure 3, when operating the stirring device 1 configured as described above, the electric motor 51 is driven to rotate the rotating body 41 in the stirring direction while the culture medium C, consisting of the culture medium and cells added to the culture medium, is contained in the annular channel.

[0034] As a result, an external force is applied to the culture medium C from the cylindrical outer surface 43 of the rotating body 41, causing the culture medium C to begin flowing in the stirring direction, similar to the rotating body 41. At this time, the annular flow path has a smaller flow path cross-sectional area in the narrowing region R1 on one side of the second direction D2 as viewed from the rotating body 41, and a larger flow path cross-sectional area in the expanding region R2 on the other side of the second direction D2. Therefore, a difference in the flow velocity of the culture medium C occurs between these narrowing region R1 and expanding region R2. That is, in the narrowing region R1, the movement speed of the culture medium C becomes relatively larger, resulting in a relatively smaller static pressure of the culture medium C. On the other hand, in the expanding region R2, the movement speed of the culture medium C becomes relatively smaller, resulting in a relatively larger static pressure.

[0035] As a result, the liquid level L1 in the contracted region R1 becomes relatively lower, while the liquid level L2 in the expanded region R2 becomes relatively higher, creating a difference in liquid level between the contracted region R1 and the expanded region R2.

[0036] Then, when the liquid level L2 in the expanded region R2 exceeds the height of the top 44a of the rotating body 41, the culture medium C in the expanded region R2 flows over the top 44a of the rotating body 41 toward the contracted region R1. At this time, since the liquid level L1 in the contracted region R1 is lower than the liquid level L2 in the expanded region R2, the culture medium C falls from the expanded region R2 toward the contracted region R1. While the electric motor 51 is running, the above liquid level difference is continuously maintained, and the flow of the culture medium C from the expanded region R2 toward the contracted region R1 also continues.

[0037] Furthermore, when the stirring device 1 is in operation, culture oxygen is supplied to the culture medium C from the gas discharge section 24. This culture oxygen and culture medium C move along the annular channel as the flow occurs, and during this movement process, the culture oxygen dissolves into the culture medium C.

[0038] Then, any culture oxygen that does not dissolve completely in the culture medium C escapes from the liquid surface L2 of the expanded region R2 into the upper space (gas phase) within the annular channel and is discharged to the outside of the external structure 10 via the air discharge section 15.

[0039] <Effects> The stirring device 1 of the first embodiment described above is equipped with an air supply unit 14 that supplies air from above to below toward the liquid surface LS of the culture medium C contained in the annular channel. This allows the air supplied by the air supply unit 14 to collide with the liquid surface LS. As a result, the liquid surface LS behaves in a wave-like manner, making it possible to dissolve oxygen from the gas phase, such as culture oxygen that has escaped into the gas phase without dissolving in the culture medium C, into the culture medium C from the liquid surface LS. Therefore, it becomes possible to promote gas-liquid contact between the liquid phase and the gas phase within the external structure 10.

[0040] Furthermore, in the stirring device 1 of the first embodiment described above, the external structure 10 comprises an external structure body 11 capable of containing the culture medium C, and a lid 12 that can open and close the upper opening 13 of the external structure body 11, with an air supply unit 14 provided on the lid 12. This makes it possible to divide the external structure 10 into upper and lower sections. Therefore, compared to the case where the external structure 10 is an indivisible container, the complexity of the manufacturing process of the external structure 10 can be suppressed.

[0041] Furthermore, in the stirring device 1 of the first embodiment described above, the culture medium C flows from the expanding region R2 to the contracting region R1 in the annular channel. This flow draws air from the upper part of the annular channel into the culture medium C. This allows bubbles to be diffused into the culture medium C and promotes the dissolution of air into the culture medium C. As a result, the oxygen concentration in the culture medium C can be increased, promoting cell growth and improving cell viability.

[0042] Furthermore, in the stirring device 1 of the first embodiment described above, the external structure 10 is positioned above the liquid surface L2 of the expanded region R2 and on the other side of the second direction D2 from the air supply unit 14, and is equipped with an air discharge unit 15 capable of exhausting air from the annular flow path. As a result, the air A1 flowing in from the air supply unit 14 flows from one side of the second direction D2 to the other side of the second direction D2 and can be exhausted from the air discharge unit 15. Therefore, since the flow is opposite to the flow of the culture medium C at the liquid surface L2, the shear force at the liquid surface L2 can be increased and gas-liquid contact can be promoted.

[0043] In addition, in the stirring device 1 of the first embodiment, air is supplied obliquely from above to below so as to form a counterflow Cf that opposes the flowing direction of the culture solution C at the liquid surface LS of the culture solution C. Thereby, while forming the counterflow Cf by the air A1 supplied by the air supply unit 14, the air A1 can be blown onto the liquid surface LS (liquid surface L2). Therefore, it is possible to further cause the behavior of the liquid surface LS (liquid surface L2), such as making the liquid surface LS (liquid surface L2) ripple.

[0044] Furthermore, in the stirring device 1 of the first embodiment, the air discharge unit 15 is provided with an antifoaming unit 16 that destroys bubbles by coming into contact with the bubbles generated in the culture solution C. Therefore, when the bubbles move along the flow of the air discharged from the air discharge unit 15, the bubbles can be destroyed and defoamed.

[0045] In addition, in the stirring device 1 of the first embodiment, the external structure 10 has a liquid discharge unit 23 that can discharge the culture solution C accommodated in the annular flow path to the outside of the external structure 10 by communicating the annular flow path with the outside of the external structure 10. Thereby, when moving the culture solution C to another container 60 or measuring the number of cells in the culture solution C, etc., there is no need to suck out the culture solution C with a pipette tip. Therefore, the troublesome operation of sucking out the culture solution with a pipette tip becomes unnecessary. Furthermore, the risk of so-called contamination can be reduced compared to the case of sucking out the culture solution with a pipette tip. Also, generally, since a pipette tip, which is a consumable, is not required, cost reduction can be achieved.

[0046] Furthermore, in the stirring device 1 of the first embodiment, a gas release unit 24 capable of releasing gas into the culture solution C in the annular flow path is provided. Thereby, oxygen can be easily supplied into the culture solution C even when the annular flow path is swirling.

[0047] <Second Embodiment> Next, the second embodiment of the present disclosure will be described based on FIG. 5. The stirring device of this second embodiment is provided with a liquid supply section with respect to the stirring device 1 of the above-described first embodiment. Therefore, the same parts as those in the above-described first embodiment will be described with the same reference numerals, and redundant descriptions will be omitted. FIG. 5 is a cross-sectional view of the stirring device in the second embodiment of the present disclosure. In FIG. 5, for reasons of illustration, the illustration of the air supply section 14 and the air discharge section 15 provided in the lid body 12 is omitted.

[0048] <Stirring Device> As shown in FIG. 5, in the second embodiment, the case of performing subculture by two stirring devices 1A and 1B will be described as an example. The configurations of the two stirring devices 1A and 1B in the second embodiment are the same. The stirring devices 1A and 1B include an external structure 10, an internal structure 40, and a drive section 50, similarly to the stirring device 1 of the above-described first embodiment. The external structure 10 includes an external structure main body 11 and a lid body 12. Further, the external structure 10 includes an air supply section 14, an air discharge section 15, and a plurality of liquid supply sections (liquid supply sections 33 to 35). Note that the number of liquid supply sections is not limited to three, and any necessary number may be provided.

[0049] The stirring devices 1A and 1B of this second embodiment include a compressed air supply section 32 capable of supplying compressed air A2 to the gas phase in the external structure 10. It is possible to increase the pressure of the gas phase by the compressed air A2 supplied by the compressed air supply section 32, and when transferring the culture solution C from the stirring device 1A to the stirring device 1B for subculture, it is possible to push out the culture solution C from the liquid discharge section 23. Note that the air for increasing the pressure of the gas phase may be supplied by the above-described air supply section 14. In this case, the compressed air supply section 32 can be omitted.

[0050] <Liquid Supply Section> The liquid supply sections 33 to 35 are formed so as to be able to supply liquid to the annular flow path. In this second embodiment, the liquid supply sections 33 to 35 are provided in the lid body 12. These liquid supply sections 33 to 35 are formed so as to be able to be closed so that the annular flow path and the external space of the external structure 10 are not communicated when not in use. Note that in FIG. 5, the case where the liquid supply sections 33 to 35 are tubular is illustrated, but the liquid supply sections 33 to 35 are not limited to being tubular.

[0051] In the subculture performed using the stirring devices 1A and 1B of this second embodiment, the first generation of culture is performed by stirring device 1A, and the second generation of culture is performed by stirring device 1B. When performing the second generation of culture with stirring device 1B, a portion of the culture medium C contained in the annular channel of stirring device 1A is transferred to the annular channel of stirring device 1B. Specifically, a portion of the culture medium C pushed out from the liquid discharge section 23 of stirring device 1A is supplied to the annular channel of stirring device 1B via the liquid supply section 35 of stirring device 1B. In addition, fresh culture medium Cn is supplied to the annular channel of stirring device 1B from the outside via the liquid supply section 34 of stirring device 1B. In addition, glucose G is supplied to the annular channel of stirring device 1B from the outside via the liquid supply section 33 of stirring device 1B. Subsequently, cell culture is performed while stirring the culture medium C in the annular channel of stirring device 1B.

[0052] In other words, the liquid supply unit 35 functions as a culture medium supply unit for supplying culture medium C, the liquid supply unit 34 functions as a culture medium supply unit for supplying the culture medium necessary for culture performed by the stirring device 1B, and the liquid supply unit 33 functions as a glucose supply unit for supplying the glucose G necessary for culture performed by the stirring device 1B. Although the culture medium supply unit and glucose supply unit have been described, a liquid supply unit (cell suspension supply unit) capable of supplying cell suspension may also be provided. Furthermore, in the second embodiment, two generations of subculturing were performed using two stirring devices 1A and 1B as an example, but three generations or more of subculturing may be performed using three stirring devices.

[0053] <Effects> According to the stirring devices 1A and 1B of the second embodiment described above, by providing multiple liquid supply sections 33 to 35, subculturing can be performed smoothly without using pipette tips. Therefore, the risk of so-called contamination during subculturing can be reduced.

[0054] <Third Embodiment> Next, a third embodiment of the present disclosure will be described with reference to Figure 6. The stirring device of this third embodiment differs from the stirring device 1 of the first embodiment described above in the configuration of the liquid discharge section 23. For this reason, the same reference numerals are used for the same parts as in the first and second embodiments described above, and redundant explanations are omitted. Also, in Figure 6, for illustrative purposes, the air supply section 14 and air discharge section 15 provided on the lid 12 are not shown.

[0055] Figure 6 is a cross-sectional view showing the schematic configuration of a stirring device in the third embodiment of this disclosure. As shown in Figure 6, the stirring device 1C of this third embodiment includes an external structure 10, an internal structure 40, and a drive unit 50, similar to the stirring devices 1, 1A, and 1B of the first and second embodiments described above. The external structure 10 includes an external structure body 11 and a lid 12. The external structure 10 also includes an air supply unit 14, an air discharge unit 15, a plurality of liquid supply units (liquid supply units 33 to 35), and a compressed air supply unit 32.

[0056] The external structure body 11 has a liquid discharge section 123 that can discharge the culture medium C from within the annular channel. The liquid discharge section 123 is tubular in shape and forms a channel for the culture medium C to be discharged to the outside of the external structure 10. The liquid discharge section 123 is also made of a material that can transmit the measurement light of the measuring device 70 in order to measure the culture medium C. Transparent synthetic resins or glass can be used as materials that can transmit the measurement light. Examples of measurements using measurement light include cell count measurement in subculture and glucose concentration measurement. Furthermore, examples of measurement methods using measurement light include semiconductor microscopy, Raman spectroscopy, and infrared light measurement. In this third embodiment, measurement is performed by Raman spectroscopy, and the liquid discharge section 123 has a tapered shape.

[0057] <Effects> According to the stirring device 1C of the third embodiment described above, the culture medium C that is pushed out from the external structure 10 and flows through the liquid discharge section 123 can be measured using measuring light. Therefore, it is possible to easily measure the culture medium C without contact, without using pipette tips or the like.

[0058] <Fourth Embodiment> Next, the fourth embodiment of the present disclosure will be described with reference to Figure 7. The stirring device of this fourth embodiment differs from that of the second embodiment described above in its purpose of use. For this reason, the same reference numerals are used for the same parts as in the first and second embodiments described above, and redundant explanations are omitted. Figure 7 is a cross-sectional view of the stirring device in the fourth embodiment of the present disclosure. In Figure 7, for illustrative purposes, the air supply unit 14 and the air discharge unit 15 provided on the lid 12 are omitted from the illustration.

[0059] <Agitation Device> As shown in Figure 7, in the fourth embodiment, we will explain an example in which culture is performed using agitation device 1A while measurement is performed using agitation device 1D. The configurations of the two agitation devices 1A and 1D in the fourth embodiment are the same. Similar to the agitation device 1 of the first embodiment described above, agitation devices 1A and 1D are equipped with an external structure 10, an internal structure 40, and a drive unit 50. The external structure 10 is equipped with an external structure body 11 and a lid 12. The external structure 10 is also equipped with an air supply unit 14, an air discharge unit 15, and a plurality of liquid supply units (liquid supply units 33 to 35). Note that the number of liquid supply units is not limited to three, and as many as necessary may be provided.

[0060] The stirring device 1A of this fourth embodiment is equipped with a compressed air supply unit 32 capable of supplying compressed air A2 to the gas phase within the external structure 10. The compressed air A2 supplied by the compressed air supply unit 32 can increase the pressure of the gas phase. When transferring the culture medium C from the stirring device 1A to the stirring device 1D, the culture medium C can be pushed out from the liquid discharge unit 23. The air used to increase the pressure of the gas phase may also be supplied by the air supply unit 14 described above. In this case, the compressed air supply unit 32 can be omitted (the same applies to the fifth embodiment below).

[0061] <Liquid Supply Section> The liquid supply sections 33 to 35 are formed to supply liquid to the annular channel. In this fourth embodiment of the stirring device 1A and 1D, the liquid supply sections 33 to 35 are provided on the lid 12, similar to the second embodiment. These liquid supply sections 33 to 35 are formed to be able to be closed when not in use so that the annular channel and the external space of the external structure 10 are not connected. Although Figure 7 illustrates the case where the liquid supply sections 33 to 35 are tubular, the liquid supply sections 33 to 35 are not limited to being tubular.

[0062] In the fourth embodiment, while the stirring device 1A performs cell culture, the stirring device 1D is used as a container for measuring the culture medium C, for example, for measuring the number of cells to determine whether the cells are viable or not. First, when performing the measurement, a portion of the culture medium C contained in the annular channel of the stirring device 1A is transferred to the annular channel of the stirring device 1D. At this time, a portion of the culture medium C pushed out from the liquid discharge section 23 of the stirring device 1A is supplied to the annular channel of the stirring device 1D via the liquid supply section 35 of the stirring device 1D.

[0063] Furthermore, a measurement solution T, such as trypan blue, a biological staining agent, is supplied from the outside to the annular channel of the stirring device 1D via the liquid supply section 34 of the stirring device 1D. Also, a measurement solution D, such as a diluent, is supplied from the outside to the annular channel of the stirring device 1D via the liquid supply section 33 of the stirring device 1D. In other words, the liquid supply sections 33 and 34 function as measurement solution supply sections for supplying the measurement solution T and measurement solution D necessary for the measurement performed by the stirring device 1D. With a portion of the culture medium C, the measurement solution T, and the measurement solution D contained in the annular channel of the stirring device 1D, when the drive unit 50 is driven in the same manner as the stirring device 1 of the first embodiment, the liquid contained in the annular channel is stirred by swirling around the rotating body 41. As a result, the culture medium C, the measurement solution T, and the measurement solution D, which are the liquids contained in the stirring device 1D, are uniformly mixed. Then, in this fourth embodiment, measurements such as the measurement of cell count are performed using the mixed liquid.

[0064] <Effects> According to the stirring devices 1A and 1D of the fourth embodiment described above, by providing multiple liquid supply sections 33 to 35, measurement work on the culture medium C, such as measuring the number of cells, can be performed smoothly without using pipette tips. Therefore, the burden on the worker involved in the measurement work of the culture medium C used for cell culture can be reduced.

[0065] <Modification of the Fourth Embodiment> Figure 8 is a cross-sectional view of the external structure body in a modification of the fourth embodiment of the present disclosure. Figure 9 is a cross-sectional view of the external structure body of Figure 8, viewed from the right. In the stirring device 1D of the fourth embodiment described above, at least a part of the external structure body 11 may be made of a transparent material so that the liquid contained inside the external structure body 11 can be imaged from outside the external structure body 11.

[0066] Furthermore, as shown in the modified fourth embodiment in Figures 8 and 9, an imaging area 80 for imaging the liquid inside the external structure body 11 may be set on the end plate 30, and an imaging recess 81 that is recessed in a first direction D1 according to the focal length of the imaging equipment may be provided in this imaging area 80. By forming such an imaging recess 81, the quality of imaging in the imaging area 80 can be improved.

[0067] <Fifth Embodiment> Next, the stirring device and subculture system in the fifth embodiment of the present disclosure will be described with reference to Figure 10. The stirring device of this fifth embodiment differs from the configuration of the stirring devices 1A, 1B, and 1D described above in that it is provided with a plurality of liquid discharge parts. For this reason, the same reference numerals are used for the same parts as in the first, second, and third embodiments described above, and redundant explanations are omitted. Figure 10 is a cross-sectional view showing the schematic configuration of the subculture system in the fifth embodiment of the present disclosure.

[0068] <Stirring device> As shown in Figure 10, in the fifth embodiment, the subculture system 100 includes multiple stirring devices 101A and 101B which perform subculture, and multiple stirring devices 1D which perform measurement. The configurations of stirring devices 101A, 101B and stirring device 1D are the same.

[0069] In this fifth embodiment, two stirring devices 101A and 101B for subculturing are illustrated, but the number of stirring devices for subculturing can be set according to the required number of generations of subculturing, and may be three or more (for example, stirring devices 101A, 101B..., 101N).

[0070] The stirring devices 101A and 101B, like the stirring device 1 of the first embodiment described above, are equipped with an external structure 10, an internal structure 40, and a drive unit 50. The external structure 10 comprises an external structure body 11 and a lid 12. The external structure 10 also includes an air supply unit 14, an air discharge unit 15, a plurality of liquid supply units (liquid supply units 33 to 35), a first liquid discharge unit 23A, a second liquid discharge unit 23B, and a gas discharge unit 24. In Figure 10, for illustrative purposes, the air supply unit 14, air discharge unit 15, and gas discharge unit 24 provided on the lid 12 are omitted from the illustration.

[0071] <Liquid Discharge Sections> The first liquid discharge section 23A and the second liquid discharge section 23B are configured to communicate the annular flow path with the outside of the external structure 10, similar to the liquid discharge section 23 of the first embodiment. The first liquid discharge section 23A and the second liquid discharge section 23B form a flow path that can discharge the culture medium C contained in the annular flow path to the outside of the external structure 10. In the fifth embodiment, the first liquid discharge section 23A is formed in the shape of a tube that extends diagonally downward from the lower part of the curved surface section 20 and opens diagonally downward. The second liquid discharge section 23B is formed in the shape of a tube that extends downward from the lower part of the curved surface section 20 and opens.

[0072] The first liquid discharge section 23A and the second liquid discharge section 23B are each provided with a first opening / closing section 25 that opens and closes the flow path within the liquid discharge section. For example, by supplying air A1 to the gas phase above the liquid level LS to increase the pressure of the gas phase and opening the first opening / closing section 25 of the first liquid discharge section 23A from a closed state, the culture medium C in the annular flow path can be pushed out to the outside of the external structure 10 via the first liquid discharge section 23A. Similarly, by increasing the pressure of the gas phase and opening the first opening / closing section 25 of the second liquid discharge section 23B from a closed state, the culture medium C in the annular flow path can be pushed out to the outside of the external structure 10 via the second liquid discharge section 23B.

[0073] Similar to the fourth embodiment, the stirring device 1D is used as a container for measuring the number of cells in each culture medium C that has been cultured by stirring devices 101A and 101B. One stirring device 1D is provided for each of the multiple stirring devices 101A and 101B that perform subculturing as described above. In other words, the multiple stirring devices 1D are provided in pairs with the stirring devices 101A and 101B that perform subculturing.

[0074] <Subculture Method> In the subculture method using the subculture system 100 of the fifth embodiment, the first generation of culture is performed by the stirring device 101A, and the second generation of culture is performed by the stirring device 101B. That is, a portion of the culture medium C contained in the annular channel of the stirring device 101A is transferred to the annular channel of the stirring device 101B. At this time, a portion of the culture medium C pushed out from the first liquid discharge section 23A of the stirring device 101A is supplied to the annular channel of the stirring device 101B via the liquid supply section 35 of the stirring device 101B. Then, as in the second embodiment, fresh culture medium Cn is supplied to the annular channel of the stirring device 101B from the outside via the liquid supply section 34 of the stirring device 101B, and glucose G is supplied to the annular channel of the stirring device 101B from the outside via the liquid supply section 33. The subculture process is carried out in this manner.

[0075] A portion of the culture medium C contained in the annular channel of the agitator 101A is transferred to the annular channel of the agitator 1D, which is paired with the agitator 101A, similar to the fourth embodiment. That is, a portion of the culture medium C is supplied to the annular channel of the agitator 1D via the second liquid discharge section 23B of the agitator 101A and the liquid supply section 32 of the agitator 1D. A portion of the culture medium C contained in the annular channel of the agitator 101B is also transferred to the annular channel of the agitator 1D, which is paired with the agitator 101B, similar to the case of the agitator 101A.

[0076] Then, in the annular channel of each stirring device 1D to which some of the culture medium C has been transferred, a measurement solution T, such as trypan blue, a biological staining agent, is supplied from the outside via the liquid supply section 34 of the stirring device 1D, similar to the fourth embodiment. In addition, a measurement solution D, such as a diluent, is supplied from the outside to the annular channel of the stirring device 1D via the liquid supply section 33 of the stirring device 1D. In other words, the liquid supply sections 33 and 34 function as measurement solution supply sections for supplying the measurement solution T and measurement solution D necessary for the measurement performed by the stirring device 1D. Subsequently, measurements such as cell count are performed using a mixture of measurement solution T, measurement solution D and culture medium C. In this way, the measurement process is performed in parallel with the subculturing process. Note that although the stirring device 1D in this fifth embodiment is shown as having the same configuration as the stirring devices 101A and 101B, it is not limited to having the same configuration as the stirring devices 101A and 101B. For example, it may have the same configuration as the stirring device 1D in the fourth embodiment or a modified version of the fourth embodiment.

[0077] <Effects> The stirring devices 101A and 101B and the subculture system 100 of the fifth embodiment described above have multiple liquid discharge sections, including a first liquid discharge section 23A and a second liquid discharge section 23B. With a subculture method using such a subculture system 100, it is possible to measure the number of cells, etc., in parallel with subculture without using pipette tips, etc. Therefore, the risk of contamination of the culture medium C can be reduced, and the burden on the operator can be reduced.

[0078] <Modification of the Fifth Embodiment> In the fifth embodiment described above, the case in which a stirring device 1D is used as the container for measurement was explained as an example, but for example, the container 60 of the first embodiment described above may be used. In this configuration as well, it is possible to measure the number of cells, etc. in parallel with subculturing, similar to the fifth embodiment.

[0079] (Other Embodiments) Although embodiments of the present disclosure have been described in detail above with reference to the drawings, the specific configuration is not limited to these embodiments and may include design changes and the like that do not depart from the gist of the present disclosure.

[0080] For example, the case described is one in which the air supply unit 14 is located vertically above the liquid surface L1 and supplies air diagonally downward. However, the arrangement of the air sharing unit 14 is not limited to vertically above the liquid surface L1, and the direction in which the air supply unit 14 supplies air is not limited to diagonal.

[0081] Furthermore, an air pump or the like may be used as the drive unit 50 to apply an external force to the culture medium C in the annular flow path using lifting air. This allows the culture medium C to flow in the stirring direction without rotating the internal structure 40, or while the internal structure 40 is rotating.

[0082] Furthermore, the cylindrical outer surface 43 of the internal structure 40 does not have to be uniform in the direction of the eccentric axis O2. For example, it may be configured such that large diameter sections and small diameter sections are alternately arranged in the direction of the eccentric axis O2. Also, spiral grooves or protrusions that twist around the eccentric axis O2 as they are directed toward the direction of the eccentric axis O2 may be formed on the cylindrical outer surface 43 of the internal structure 40.

[0083] The cylindrical outer surface 43 of the internal structure 40 does not have to be perfectly circular in shape perpendicular to the eccentric axis O2, but may be elliptical, polygonal, or the like. The inner surface of the external structure 10 does not have to be perfectly circular in shape perpendicular to the axis, but may be elliptical, polygonal, or the like.

[0084] Even in these cases, by positioning the internal structure 40 so as to be offset horizontally to one side relative to the external structure 10, a reduced region R1 and an expanded region R2 can be formed in the annular channel. Furthermore, by positioning the internal structure 40 so as to be offset upward relative to the external structure 10, the capacity of the culture medium C within the annular channel can be increased.

[0085] The internal structure 40 does not need to be positioned offset upward relative to the external structure 10; in other words, the eccentric axis O2 may be at the same height as the central axis O1.

[0086] Furthermore, although the example given shows the air supply unit 14 as a hole, the air supply unit 14 may also be tubular in shape, penetrating the external structure 10 from the inside to the outside.

[0087] Furthermore, a case was described in which a counterflow Cf opposite to the flow of the culture medium C at the liquid surface LS(L2) is generated by air supplied by the air supply unit 14. However, it is not limited to a counterflow Cf, and a parallel flow along the flow of the culture medium C at the liquid surface LS(L2) may also be used. In this case, the functions of the air supply unit 14 and the air discharge unit 15 may be interchangeable. That is, air may be supplied from the air discharge unit 15 and air may be discharged from the air supply unit 14.

[0088] Furthermore, in the above embodiment, the external structure 10 is described as being composed of an external structure body 11 and a lid 12, and the external structure body 11 can be opened and closed by the lid 12. However, the external structure body 11 and the lid 12 may be integrated and configured so that they cannot be opened and closed.

[0089] In the fifth embodiment, the stirring devices 101A and 101B were provided with two liquid discharge sections 23 as a plurality of liquid discharge sections 23. However, the number of liquid discharge sections 23 can be provided according to the number of destinations for the liquid discharge. For example, three or more liquid discharge sections 23 may be provided for a single stirring device.

[0090] <Note> The stirring device 1 and subculture system 100 described in each embodiment can be understood, for example, as follows.

[0091] (1) According to the first embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, and 101B each include a hollow external structure 10, an internal structure 40 disposed within the external structure 10 and defining an annular channel containing culture medium C and air between itself and the inner circumferential surface 22 of the external structure 10, and a drive unit 50 that applies an external force to the culture medium C in the annular channel in the circumferential direction of the annular channel, wherein the external structure 10 includes an air supply unit 14 that supplies air from above downward toward the liquid surface LS of the culture medium C contained in the annular channel.

[0092] This allows the air supplied by the air supply unit 14 to collide with the liquid surface LS. This causes the liquid surface LS to ripple or exhibit other behaviors, making it possible to dissolve oxygen from the gas phase, such as culture oxygen that escaped into the gas phase without dissolving in the culture medium C, into the culture medium C from the liquid surface LS. Therefore, it becomes possible to promote gas-liquid contact between the liquid phase and the gas phase within the external structure 10.

[0093] (2) According to the second embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B of (1), wherein the external structure 10 comprises an external structure body 11 capable of containing at least the culture medium C and having an upper opening 13 that opens upward, and a lid 12 that can open and close the upper opening 13 of the external structure body 11, and the air supply unit 14 is provided on the lid 12.

[0094] This makes it possible to divide the external structure 10 into upper and lower sections. Therefore, compared to the case where the external structure 10 is an indivisible container, the complexity of the manufacturing process for the external structure 10 can be minimized.

[0095] (3) According to the third embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B of (1), wherein the annular flow path has a reduced region R1 which is the region on one side of the second direction D2 which is the horizontal direction perpendicular to the first direction D1 in which the internal structure 40 extends, and an expanded region R2 which is the region on the other side of the second direction D2 of the internal structure 40 and has a larger flow path cross-sectional area than the reduced region R1, the drive unit 50 applies the circumferential external force to the culture medium C in the annular flow path from the expanded region R2 toward the reduced region R1 above the internal structure 40, and the air supply unit 14 supplies the air toward the liquid surface LS of the expanded region R2.

[0096] This allows bubbles to diffuse into the culture medium C and promotes the dissolution of air into the culture medium C. As a result, the oxygen concentration in the culture medium C can be increased, promoting cell growth and improving cell viability.

[0097] (4) According to the fourth embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are the stirring device 1 of (1), wherein the external structure 10 further includes an air discharge unit 15 capable of exhausting air from the annular flow path, located above the liquid surface LS of the expanded region R2 and on the other side of the air supply unit 14 in the second direction D2.

[0098] As a result, the air A1 flowing in from the air supply unit 14 flows from one side of the second direction D2 to the other side of the second direction D2 and can be exhausted from the air discharge unit 15. Therefore, since the flow is opposite to the flow of the culture medium C at the liquid surface L2, the shear force at the liquid surface L2 can be increased and gas-liquid contact can be promoted.

[0099] (5) According to the fifth embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B of (1), and the air supply unit 14 supplies the air diagonally from above the reduced area onto the liquid surface of the expanded area.

[0100] This allows the air A1 supplied by the air supply unit 14 to create a counterflow Cf that is opposed to the flow of liquid surfaces LS and L2, while simultaneously blowing the air A1 onto the liquid surfaces LS and L2. Therefore, it becomes possible to create even more behavior in the liquid surfaces LS and L2, such as causing waves in them.

[0101] (6) According to the sixth embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B of (1) to (5), and the air discharge unit 15 is equipped with a defoaming unit 16 for destroying foam.

[0102] As a result, when bubbles move along the airflow discharged from the air discharge section 15, the bubbles can be destroyed and defoamed.

[0103] (7) According to the seventh embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B of (1), wherein the external structure 10 further comprises liquid discharge sections 23, 23A, 23B that connect the annular flow path to the outside of the external structure 10 and allow the culture medium C contained in the annular flow path to be discharged to the outside of the external structure 10.

[0104] This eliminates the need to pipette the culture medium C when transferring it to another container 60 or when measuring the number of cells in the culture medium C. Therefore, the cumbersome process of pipetteting the culture medium C is eliminated. Furthermore, it reduces the risk of contamination compared to when pipetteting the culture medium C. In addition, since it does not require pipette tips, which are generally consumables, it is possible to reduce costs.

[0105] (8) According to the eighth aspect, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B of (1), further comprising a first opening / closing section 25 for opening and closing the liquid discharge sections 23, 23A, 23B.

[0106] This allows air A1 to be supplied to the gas phase above the liquid level LS, increasing the pressure of the gas phase, and by opening the first opening / closing section 25 from its closed state, the culture medium C in the annular channel can be easily pushed out through the liquid discharge sections 23, 23A, and 23B to a container 60 or the like located outside the external structure 10.

[0107] (9) According to the ninth embodiment, the stirring device 1, 1A, 1B, 1C, 1D, 101A, 101B is any one of the stirring devices 1 from (1) to (8), and the external structure 10 further comprises a gas release section 24 capable of releasing gas into the culture medium C of the annular flow path.

[0108] This allows oxygen to be easily supplied to the culture medium C even when the annular channel is swirling.

[0109] (10) According to the tenth embodiment, the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B are any one of the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B from (1) to (9), and the liquid discharge sections 23, 23A, 23B, 123 are tubular in shape that form a flow path for the culture medium C discharged to the outside of the external structure 10, and are made of a material that can transmit measuring light for measuring the culture medium C.

[0110] This makes it possible to easily measure the culture medium C in a non-contact manner without using pipette tips or the like.

[0111] (11) According to the eleventh embodiment, the stirring devices 101A, 101B are any one of the stirring devices 101A, 101B from (1) to (10), and the external structure 10 has a plurality of liquid discharge sections 23A, 23B.

[0112] This allows the liquid in the annular channel to be discharged to multiple outlets without the need for pipette tips or the like. Furthermore, cell counts and other parameters can be measured in parallel with subculturing using the stirring device 101A or stirring device 101B.

[0113] (12) According to the twelfth embodiment, the stirring device 1C is any one of the stirring devices 1C from (1) to (11), and the liquid discharge section 123 has a tapered shape.

[0114] This makes it possible to easily measure the culture medium C in a non-contact manner without using pipette tips or the like.

[0115] (13) According to the 13th embodiment, the stirring device 1A, 1B, 1C, 1D, 101A, 101B is any one of the stirring devices 1 of (1) to (12), and the external structure 10 further includes at least one of the following as liquid supply units 33, 34, 35 for supplying liquid to the annular channel: a culture medium supply unit for supplying culture medium to the annular channel, a glucose supply unit for supplying glucose to the annular channel, a cell suspension supply unit for supplying cell suspension to the annular channel, and a measurement liquid supply unit for supplying measurement liquid to the annular channel.

[0116] This allows liquids necessary for cell culture and measurement to be supplied to the annular channel without the use of pipette tips. Therefore, the risk of contamination can be reduced.

[0117] (14) According to the 14th embodiment, the stirring device 1 is any one of the stirring devices 1 of (1) to (13), wherein the external structure 10 further has a recess 81 formed of a material that allows the culture medium C in the annular channel to be imaged from the outside and recesses in the first direction D1.

[0118] This makes it possible to improve the quality of imaging in the imaging area 80.

[0119] (15) According to the 15th embodiment, the subculture system 100 is a subculture system 100 using the stirring devices 1D, 101A, and 101B of (10), comprising, as a plurality of stirring devices, first stirring devices 101A and 101B for performing subculture, and second stirring device 1D for measuring the culture medium C, wherein the second stirring device 1D is equipped with measuring liquid supply units 33 and 34 capable of supplying measuring liquids T and D for measurement to the annular flow path, and a plurality of liquid discharge units 23 provided in one of the first stirring devices 1A and 101A include a first liquid discharge unit 23A that communicates with the annular flow path of adjacent first stirring devices 1B and 101B, and a second liquid discharge unit 23B that communicates with the annular flow path of the second stirring device 1D.

[0120] This allows for the measurement of cell count and other parameters in parallel with subculturing, without the need for pipette tips or similar devices. Therefore, the risk of contamination of culture medium C can be reduced, and the burden on the operator can be lessened.

[0121] (16) According to the 16th embodiment, the subculture system 100 is a subculture system 100 using the stirring devices 1A, 1B, 1D, 101A, 101B of (10), wherein the plurality of stirring devices include first stirring devices 101A, 101B for subculture, and the plurality of liquid discharge sections 23 provided in one of the first stirring devices 101A include a first liquid discharge section 23A that communicates with the annular flow path of an adjacent first stirring device 101B, and a second liquid discharge section 23B that communicates with the storage space of a measuring container 60 capable of containing the culture medium C.

[0122] (17) According to the 17th aspect, the subculture method is a subculture method using the subculture system of (15), comprising: a subculture step of sequentially performing subculture using a plurality of first stirring devices 101A, 101B connected via the first liquid discharge unit 23A; and a measurement step of supplying a portion of the culture medium C of each generation cultured by the plurality of first stirring devices 101A, 101B via the second liquid discharge unit 23B to the annular channel of the second stirring device 1D corresponding to each generation, in parallel with the subculture step, and measuring the culture medium C.

[0123] According to the subculture system of the 16th embodiment and the subculture method of the 17th embodiment, the number of cells and other parameters can be measured in parallel with subculture without using pipette tips or the like. Therefore, the risk of contamination of the culture medium C can be reduced, and the burden on the operator can be reduced.

[0124] According to this disclosure, it is possible to promote gas-liquid contact between the liquid phase and the gas phase.

[0125] 1, 1A, 1B, 1C, 1D Agitator 10 External structure 11 External structure body 12 Lid 13 Upper opening 14 Air supply section 15 Air discharge section 16 Defoaming section 20 Curved section 20ta First end 20tb Second end 21 Arc outer surface 22 Arc inner surface 23 Liquid discharge section 24 Gas discharge section 30 End plate 32 Compressed air supply section 33-35 Liquid supply section 40 Internal structure 41 Rotating body 43 Cylindrical outer surface 44a Top 44b Bottom 50 Drive unit 51 Electric motor 51a Electric motor body 51b Output shaft 60 Container 70 Measuring device 80 Imaging area 81 Imaging recess 100 Subculture system 101A, 101B Stirring device O1 Axis O2 Eccentric axis C Culture solution LS, L1, L2 Liquid level T, D Measurement solution

Claims

1. A stirring device comprising: a hollow external structure; an internal structure disposed within the external structure and defining an annular channel containing culture medium and air between itself and the inner circumferential surface of the external structure; and a drive unit that applies an external force to the culture medium in the annular channel in the circumferential direction of the annular channel, wherein the external structure includes an air supply unit that supplies air from above downward toward the liquid surface of the culture medium contained in the annular channel.

2. The stirring device according to claim 1, wherein the external structure comprises at least an external structure body capable of containing the culture medium and having an upper opening that opens upward, and a lid that can open and close the upper opening of the external structure body, and the air supply unit is provided on the lid.

3. The stirring device according to claim 1, wherein the annular channel has a contraction region which is a region on one side of a second direction which is a horizontal direction perpendicular to the first direction in which the internal structure extends, and an expansion region which is a region on the other side of the second direction of the internal structure and has a larger channel cross-sectional area than the contraction region, the drive unit applies an external force in the circumferential direction from the expansion region toward the contraction region above the internal structure, and the air supply unit supplies the air toward the liquid surface of the expansion region.

4. The stirring device according to claim 3, further comprising an air discharge section capable of exhausting air from the annular flow path, located above the external structure opposite to the liquid surface of the enlarged region and on the other side in the second direction from the air supply section.

5. The stirring device according to claim 3, wherein the air supply unit supplies the air diagonally from above the reduced area onto the liquid surface of the expanded area.

6. The stirring device according to claim 4, wherein the air discharge section is further provided with a defoaming section for destroying foam.

7. The stirring device according to claim 1, further comprising a liquid discharge section that connects the annular channel with the outside of the external structure and is capable of discharging the culture solution contained in the annular channel to the outside of the external structure.

8. The stirring device according to claim 7, further comprising a first opening / closing section for opening and closing the liquid discharge section.

9. The stirring device according to claim 1, further comprising a gas release section in the annular channel that can release gas into the culture medium.

10. The stirring device according to claim 7, wherein the liquid discharge section is tubular in shape, forming a flow path for the culture medium to be discharged to the outside of the external structure, and is made of a material that can transmit measuring light for measuring the culture medium.

11. The stirring device according to claim 8, wherein the external structure has a plurality of liquid discharge sections.

12. The stirring device according to claim 10, wherein the liquid discharge section has a tapered shape.

13. The stirring device according to claim 1, wherein the external structure further comprises at least one of the following as a liquid supply unit for supplying liquid to the annular channel: a culture medium supply unit for supplying culture medium to the annular channel; a glucose supply unit for supplying glucose to the annular channel; a cell suspension supply unit for supplying cell suspension to the annular channel; and a measurement liquid supply unit for supplying measurement liquid to the annular channel.

14. The stirring device according to claim 1, wherein the external structure is formed of a material that allows imaging of the culture solution in the annular channel from the outside and further has a recess that is recessed in the first direction.

15. A subculture system using the stirring device described in claim 11, comprising: a plurality of first stirring devices for performing subculture; and a second stirring device for measuring the culture medium, wherein the second stirring device is equipped with a measuring liquid supply unit capable of supplying a measuring liquid for measurement to the annular flow path; and a plurality of liquid discharge units provided in one of the first stirring devices include a first liquid discharge unit that communicates with the annular flow path of an adjacent first stirring device and a second liquid discharge unit that communicates with the annular flow path of the second stirring device.

16. A subculture system using the stirring device described in claim 11, comprising a plurality of stirring devices, the first stirring device for performing subculture, wherein a plurality of liquid discharge sections provided in one of the first stirring devices include a first liquid discharge section that communicates with the annular flow path of adjacent first stirring devices, and a second liquid discharge section that communicates with the storage space of a measuring container capable of containing the culture solution.

17. A subculture method using the subculture system described in claim 15, comprising: a subculture step of sequentially performing subculture using a plurality of first stirring devices connected via the first liquid discharge unit; and a measurement step of supplying a portion of the culture medium of each generation cultured by the plurality of first stirring devices via the second liquid discharge unit to the annular channel of the second stirring device corresponding to each generation, in parallel with the subculture step, and measuring the culture medium.

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