Stirring device, stirring system, and subculture method

Abstract

The present invention provides a stirring device and stirring system capable of promoting gas-liquid contact between a liquid phase and a gas phase. [Solution] The invention comprises a hollow external structure, an internal structure disposed within the external structure and partitioning 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.

Description

Technical Field

[0001] The present disclosure relates to a stirring device, a stirring system, and a subculture method.

Background Art

[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 into the annular flow path from below. 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.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[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, it may not be possible to maintain the culture solution 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.

Means for Solving the Problems

[0006] To solve the above problems, the stirring device according to the present disclosure comprises a hollow external structure, an internal structure disposed within the external structure and partitioning 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.

[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. [Effects of the Invention]

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

[0010] [Figure 1] This is a cross-sectional view showing the schematic configuration of a stirring device in the first embodiment of the present disclosure. [Figure 2] This is a cross-sectional view of the stirring device shown in Figure 1 above, viewed from the right. [Figure 3]This is a cross-sectional view showing the state in which culture is being carried out using the culture medium with the above stirring device. [Figure 4] This is a cross-sectional view showing the state in which the culture medium is being removed from the above stirring device. [Figure 5] This is a cross-sectional view of the stirring device in the second embodiment of the present disclosure. [Figure 6] This is a cross-sectional view showing the schematic configuration of the stirring device in the third embodiment of this disclosure. [Figure 7] This is a cross-sectional view of a stirring device in the fourth embodiment of the present disclosure. [Figure 8] This is a cross-sectional view of the external structure body in a modified example of the fourth embodiment of the present disclosure. [Figure 9] This is a cross-sectional view of the main body of the external structure shown in Figure 8 above, viewed from the right. [Figure 10] This is a cross-sectional view showing the schematic configuration of a subculturing system in the fifth embodiment of this disclosure. [Modes for carrying out the invention]

[0011] <First Embodiment> The first embodiment of this disclosure will be described in detail below 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 this 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. <Agitation 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 structures> 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] <Outer structure body> The outer structure body 11 is a container capable of storing liquid inside, and has an upper opening that opens upward. The outer structure body 11 has a curved surface portion 20 and a pair of end plates 30. The curved surface portion 20 is formed in an arc shape centered on an axis O1 extending in a first direction D1 that is a horizontal direction. The outer peripheral surface of the curved surface portion 20 is an arc outer peripheral surface 21 that forms an arc shape centered on the axis O1. The inner peripheral surface of the curved surface portion 20 is an arc inner peripheral surface 22 that forms an arc shape centered on the axis O1. The curved surface portion 20 is formed in a plate shape with a constant thickness having the radial direction centered on the axis O1 as the thickness direction.

[0014] The curved surface portion 20 has a first end portion 20ta and a second end portion 20tb in the circumferential direction centered on the axis O1. The first end portion 20ta and the second end portion 20tb form the uppermost end portion of the curved surface portion 20. The first end portion 20ta and the second end portion 20tb are arranged to be separated from each other in a second direction D2 that is a horizontal direction orthogonal to the first direction D1.

[0015] The pair of end plates 30 are provided so as to sandwich the curved surface 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 surface portion 20 in the first direction D1. A storage space inside the outer structure body 11 is formed by the pair of end plates 30 and the curved surface portion 20. Further, the upper edges of the pair of end plates 30 extend in the second direction D2, and form an upper opening 13 that opens upward together with the first end portion 20ta and the second end portion 20tb.

[0016] <Cover body> The cover body 12 is formed so as to be able to open and close the upper opening 13 of the outer structure body 11. The structure for opening and closing the cover body 12 can be appropriately selected, and examples include a screw-in type and a hinge type. The cover body 12 of the present embodiment includes an air supply portion 14 and an air discharge portion 15, which will be described later.

[0017] <Inner 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 such that it is shifted to one side of the second direction D2 compared 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 referred to as the reduced region R1 of the annular flow path. The reduced region R1 is the portion of the region within the vertical range D3 of the rotating body 41 in the annular flow path that is on one side of the eccentric axis O2 in the second direction D2.

[0020] Because the rotating body 41 is positioned biased towards one side of 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 with an enlarged flow channel cross-sectional area is referred to as the enlarged region R2 of the annular flow channel. The enlarged region R2 is the portion of the region within the vertical range D3 of the rotating body 41 in the annular flow path 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 is connected to the rotating body 41 by passing through the end plate 30. 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. Therefore, 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. In this embodiment, the air supply unit 14 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 downwards 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. The air supply unit 14 in this embodiment 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 downwards 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 flowing 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 destination (not shown) to which the air discharge section 15 is connected 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 part> 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 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 surface 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 with the above configuration, the rotating body 41 is rotated in the stirring direction by driving the electric motor 51 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 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 and Effects> The stirring device 1 of the first embodiment described above is equipped with 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. 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 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 for the external structure 10 can be minimized.

[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 causes the air at the top of the annular channel to be drawn into the culture medium C. This allows air 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.

[0042] Furthermore, in the stirring device 1 of the first embodiment described above, the external structure 10 is located above the liquid surface L2 of the expanded region R2 and is positioned 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] Furthermore, in the stirring device 1 of the first embodiment described above, air is supplied diagonally from above to below so that it forms a counterflow Cf opposite to the direction of flow of the culture medium C at the liquid surface LS of the culture medium C. This allows the air A1 supplied by the air supply unit 14 to form the counterflow Cf while simultaneously blowing the air A1 onto the liquid surface LS (liquid surface L2). Therefore, it becomes possible to create even more behavior on the liquid surface LS (liquid surface L2), such as causing waves on the liquid surface LS (liquid surface L2).

[0044] Furthermore, the stirring device 1 of the first embodiment described above is equipped with an anti-foaming unit 16 that destroys foam generated in the culture medium C by contacting the air discharge unit 15. Therefore, when bubbles move along the airflow discharged from the air discharge section 15, the bubbles can be destroyed and defoamed.

[0045] Furthermore, in the stirring device 1 of the first embodiment described above, the external structure 10 has a liquid discharge section 23 that connects the annular flow path with the outside of the external structure 10 and can discharge the culture medium C contained in the annular flow path to the outside of the external structure 10. 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 is eliminated. Furthermore, it reduces the risk of contamination compared to pipetteting the culture medium. In addition, since it does not require pipette tips, which are generally consumables, it is possible to reduce costs.

[0046] Furthermore, the stirring device 1 of the first embodiment described above has a gas release section 24 that can release gas into the culture medium C in the annular flow path. This allows oxygen to be easily supplied to the culture medium C even when the annular channel is swirling.

[0047] <Second Embodiment> Next, a second embodiment of the present disclosure will be described with reference to Figure 5. The stirring device of this second embodiment is provided with a liquid supply section in addition to the stirring device 1 of the first embodiment described above. For this reason, the same reference numerals are used for the same parts as in the first embodiment described above, and redundant explanations are omitted. Figure 5 is a cross-sectional view of the stirring device in the second embodiment of the present disclosure. In Figure 5, for illustrative purposes, the air supply section 14 and the air discharge section 15 provided on the lid 12 are not shown.

[0048] <Stirring device> As shown in Figure 5, the second embodiment will be explained using the case where subculturing is performed using two stirring devices 1A and 1B 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 are equipped with an external structure 10, an internal structure 40, and a drive unit 50, similar to the stirring device 1 of the first embodiment described above. 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.

[0049] The stirring devices 1A and 1B of this second embodiment are 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, and when transferring the culture medium C from the stirring device 1A to the stirring device 1B for subculturing, it is possible to push the culture medium C 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.

[0050] <Liquid supply section> The liquid supply sections 33-35 are formed to supply liquid to the annular channel. In this second embodiment, the liquid supply sections 33-35 are provided on the lid 12. These liquid supply sections 33-35 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 5 illustrates a case where the liquid supply sections 33-35 are tubular, the liquid supply sections 33-35 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 carried out by stirring device 1A, and the second generation of culture is carried out 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 carried out 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 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 and 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 a third embodiment of the present disclosure. As shown in Figure 6, the stirring device 1C of this third embodiment, like the stirring devices 1, 1A, and 1B of the first and second embodiments described above, is 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), 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 and Effects> According to the stirring device 1C of the third embodiment described above, the culture medium C, which 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 non-contact without using a pipette tip or the like.

[0058] <Fourth Embodiment> Next, a 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 not shown.

[0059] <Stirring device> As shown in Figure 7, the fourth embodiment will be explained using the case where culture is performed using the stirring device 1A while measurement is performed using the stirring device 1D as an example. The configurations of the two stirring devices 1A and 1D in the fourth embodiment are the same. The stirring devices 1A and 1D are equipped with an external structure 10, an internal structure 40, and a drive unit 50, similar to the stirring device 1 of the first embodiment described above. 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 33-35 are formed to supply liquid to the annular channel. In this fourth embodiment of the stirring device 1A,1D, the liquid supply sections 33-35 are provided on the lid 12, similar to the second embodiment. These liquid supply sections 33-35 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 33-35 are tubular, the liquid supply sections 33-35 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 cell viability. First, in order to perform 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 liquid 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. In addition, a measurement liquid 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 liquid supply sections for supplying the measurement liquid T and measurement liquid D necessary for measurements performed by the stirring device 1D. With a portion of the culture medium C, measurement solution T, and measurement solution D contained in the annular channel of the stirring device 1D, the drive unit 50 is driven in the same manner as in the stirring device 1 of the first embodiment, causing the liquid contained in the annular channel to swirl around the rotating body 41 and be stirred. As a result, the liquids contained in the stirring device 1D, namely the culture medium C, measurement solution T, and measurement solution D, are uniformly mixed. In this fourth embodiment, measurements such as cell count are then performed using the mixed liquid.

[0064] <Effects and 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 cell count measurement, 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] <Variation of the fourth embodiment> Figure 8 is a cross-sectional view of the external structure body in a modified example of the fourth embodiment of this 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 portion 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 this disclosure will be described with reference to Figure 10. The stirring device of this fifth embodiment differs from the stirring devices 1A, 1B, and 1D described above in that it is provided with multiple liquid discharge sections. 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 this disclosure.

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

[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 agitators 101A and 101B, like the agitator 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 33-35), a first liquid discharge unit 23A, a second liquid discharge unit 23B, and a gas discharge unit 24. Note that 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 part> 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 a tubular shape 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 a tubular shape 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 cultured by stirring devices 101A and 101B. One stirring device 1D is provided for each of the multiple stirring devices 101A and 101B used for subculturing as described above. In other words, the multiple stirring devices 1D are provided in pairs with the stirring devices 101A and 101B used for 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 agitator 101A, and the second generation of culture is performed by the agitator 101B. That is, 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 101B. At this time, a portion of the culture medium C pushed out from the first liquid discharge section 23A of the agitator 101A is supplied to the annular channel of the agitator 101B via the liquid supply section 35 of the agitator 101B. Then, as in the second embodiment, fresh culture medium Cn is supplied to the annular channel of the agitator 101B from the outside via the liquid supply section 34 of the agitator 101B, and glucose G is supplied to the annular channel of the agitator 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. In this fifth embodiment, the stirring device 1D is shown as having the same configuration as stirring devices 101A and 101B, but it is not limited to having the same configuration as 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 and Effects> The stirring devices 101A and 101B and the subculture system 100 of the fifth embodiment described above have a first liquid discharge section 23A and a second liquid discharge section 23B as multiple liquid discharge sections. This subculture method using the subculture system 100 allows for the measurement of cell count and other parameters in parallel with subculture without the need for 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.

[0078] <Modified form 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 this 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 this 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 being 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, a helical groove or protrusion may be formed on the cylindrical outer surface 43 of the internal structure 40, twisting around the eccentric axis O2 as it moves in the direction of the eccentric axis O2.

[0083] The cylindrical outer surface 43 of the internal structure 40 does not have to be a perfect circle in shape perpendicular to the eccentric axis O2; for example, it may be an ellipse or a polygon. The inner surface of the external structure 10 does not have to be a perfect circle in shape perpendicular to the axis; for example, it may be an ellipse or a polygon.

[0084] Even in these cases, by positioning the internal structure 40 so as to be off-center 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 flow 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 from 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 cover 12, and the external structure body 11 can be opened and closed by the cover 12. However, the external structure body 11 and the cover 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 comprise 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. The external structure 10 also comprises 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 behave in a wave-like manner, 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 contraction 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 expansion 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 contraction region R1, the drive unit 50 applies the circumferential external force to the culture medium C in the annular flow path from the expansion region R2 toward the contraction region R1 above the internal structure 40, and the air supply unit 14 supplies the air toward the liquid surface LS of the expansion region R2.

[0096] This allows for the diffusion of air bubbles 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 devices 1 of (1), wherein the external structure 10 further comprises 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 on the 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 on the liquid surfaces LS and L2, such as causing waves on 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 unit 25 for opening and closing the liquid discharge units 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 aspect, 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 device 1, 1A, 1B, 1C, 1D, 101A, 101B is any one of the stirring devices 1, 1A, 1B, 1C, 1D, 101A, 101B from (1) to (9), and the liquid discharge section 23, 23A, 23B, 123 is tubular in shape, forming a flow path for the culture medium C discharged to the outside of the external structure 10, and is 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 without contact, without using pipette tips or other tools.

[0111] (11) According to the eleventh embodiment, the stirring device 101A, 101B is 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 agitator 101A or agitator 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 without contact, without using pipette tips or other tools.

[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 has 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 improves the image quality 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 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 aspect, 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, a 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, cell counts 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. [Explanation of symbols]

[0124] 1,1A,1B,1C,1D Stirring device 10 External structures 11 External structure body 12 Lid 13 Top opening 14 Air supply unit 15 Air discharge section 16 Defoaming section 20 Curved section 20ta first end 20tb second end 21 Outer surface of a circular arc 22 Inner surface of a circular arc 23 Liquid discharge part 24 Gas release section 30 End plate 32 Compressed air supply unit 33~35 Liquid supply part 40 Internal structure 41. Solids of revolution 43 Cylindrical outer surface 44a Top 44b bottom 50 Drive unit 51 Electric motor 51a Motor body 51b Output shaft 60 containers 70 Measuring devices 80 imaging area 81 Imaging recess 100-passage culture system 101A,101B Stirring device O1 axis O2 eccentric axis C Culture solution LS,L1,L2 liquid level T,D measuring solution

Claims

1. The external structure is hollow, An internal structure is disposed within the external structure and separates an annular channel containing culture medium and air between itself and the inner circumferential surface of the external structure, A drive unit that applies an external force in the circumferential direction of the annular channel to the culture medium in the annular channel, Equipped with, The external structure is an agitator equipped with an air supply unit that supplies air from above to below toward the liquid surface of the culture medium contained in the annular channel.

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

3. The aforementioned annular channel is A reduced region is a region on one side of the second direction, which is a horizontal direction perpendicular to the first direction in which the internal structure extends, The internal structure has an expanded region on the other side of the second direction, where the flow path cross-sectional area is larger than that of the reduced region, The drive unit applies the circumferential external force to the culture medium in the annular channel, above the internal structure, from the expanding region to the contracting region. The air supply unit supplies the air toward the liquid surface of the enlarged area. The stirring device according to claim 1.

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

5. The aforementioned air supply unit is The air is supplied diagonally from above the reduced area onto the liquid surface of the expanded area. The stirring device according to claim 3.

6. The aforementioned air discharge section includes a defoaming section for destroying foam. The stirring device according to claim 4.

7. The aforementioned external structure is The annular channel is connected to the outside of the external structure, and the liquid discharge section is further provided to discharge the culture solution contained in the annular channel to the outside of the external structure. The stirring device according to claim 1.

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

9. The aforementioned external structure is The annular channel further comprises a gas release section capable of releasing gas into the culture medium. The stirring device according to claim 1.

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

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

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

13. The aforementioned external structure serves as a liquid supply section for supplying liquid to the annular channel, A culture medium supply unit that supplies culture medium to the aforementioned annular channel, A glucose supply unit that supplies glucose to the aforementioned annular channel, A cell suspension supply unit that supplies a cell suspension to the aforementioned annular channel, A measuring liquid supply unit that supplies measuring liquid to the aforementioned annular channel, It further has at least one of the following: The stirring device according to claim 1.

14. The aforementioned external structure is The culture medium within the annular channel is further formed of a material that can be imaged from the outside and has a recess that is recessed in the first direction. The stirring device according to claim 1.

15. A subculture system using a stirring device according to claim 11, The aforementioned stirring device comprises multiple first stirring devices for performing subculturing and a second stirring device for measuring the culture medium. The second stirring device is equipped with a measuring liquid supply unit capable of supplying a measuring liquid to the annular flow path, The plurality of liquid discharge sections provided in the first stirring device include a first liquid discharge section that communicates with the annular flow path of an adjacent first stirring device, and a second liquid discharge section that communicates with the annular flow path of the second stirring device. Subculture system.

16. A subculture system using a stirring device according to claim 11, The aforementioned stirring device includes a first stirring device for performing subculturing, The 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 medium. Subculture system.

17. A method for subculturing using the subculturing system described in claim 15, A subculture step in which subculture is carried out sequentially by a plurality of first stirring devices connected via the first liquid discharge section, The measurement step includes, in parallel with the subculturing step, a portion of the culture medium from each generation of cells cultured by a plurality of first stirring devices, supplied via the second liquid discharge section to the annular channel of the second stirring device corresponding to each generation, and measuring the culture medium. Subculture method.

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