Dispensing preparation device, dispensing device, and dispensing preparation method

Abstract

It is desirable that a dispensing preparation device capable of stirring a cell suspension without damaging cells. The problem is solved by a dispensing preparation device provided with: a reservoir that accommodates a cell suspension and has an opening from which a discharge tip of a pipette that discharges the cell suspension into a cell accommodating part repeatedly enters and exits; a tank for accommodating the cell suspension; a pipe for connecting the tank and the reservoir and allowing the cell suspension to flow from the tank to the reservoir through a pump unit; a rotary table that rotates the tank about a rotation axis perpendicular to the bottom surface of the tank; and a controller that controls the rotation of the rotary table, the pipe being disposed such that one end opening of the pipe is in contact with the inner wall of the reservoir, and the other end opening of the pipe is separated from the inner wall of the tank.

Description

Technical Field

[0001] The present invention relates to a cell suspension dispensing preparation device for dispensing a cell suspension, and a dispensing device and a dispensing preparation method using the dispensing preparation device. Background Art

[0002] In the culture of cells for preparing spheroids or the like formed by aggregating single cells, cells to be cultured are added to each well of a well plate and cultured in a CO 2 incubator. In the previous stage, a cell suspension containing the aggregated cells needs to be dispensed into each well of the well plate.

[0003] For example, Patent Documents 1 and 2 disclose a dispensing device and an accompanying device, wherein a cell suspension storage unit is directly connected to a pipette for dispensing the cell suspension to a plate, and the cell suspension can be continuously and repeatedly dispensed to the plate.

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: Japanese Patent No. 6906773

[0007] Patent Document 2: Japanese Patent Application Publication No. 2021-000018 Summary of the invention

[0008] Problems to be solved by the invention

[0009] When automating the process of dispensing a cell suspension containing cells to be cultured into each well, it is necessary to uniformly contain cells without deviation in the suspension dispensed into each well, and not to damage the cells. Regarding these points, Patent Document 1 does not disclose either uniformly distributing cells without deviation in the suspension or not damaging the cells at the dispensing stage. In addition, in the dispensing device disclosed in Patent Document 2, although a stirring device for the suspension is mentioned, a countermeasure for not damaging the cells is not disclosed.

[0010] In general, “stirring” is a representative action for homogenizing a substance in the entire area of ​​a medium containing a substance, not limited to cells. However, “stirring” is accompanied by the flow of the substance in the medium. Therefore, in order to homogenize cells in a suspension containing cells, the application of stirring may cause direct damage to the cells. Therefore, it is difficult to simply apply “stirring” in the process of dispensing cells. Therefore, a dispensing device that can stir a cell suspension in a manner that does not damage the cells is desired.

[0011] Means for solving problems

[0012] One embodiment of the present invention is a dispensing preparation device, which comprises: a reservoir that stores a cell suspension and has an opening, and a discharge tip of a pipette that discharges the cell suspension to a cell storage portion repeatedly enters and exits from the above-mentioned opening; a tank that stores the above-mentioned cell suspension; a pipe that connects the above-mentioned tank and the above-mentioned reservoir, and causes the cell suspension to flow from the above-mentioned tank to the above-mentioned reservoir internally through a pump unit; a rotating table that rotates the above-mentioned tank around a rotation axis perpendicular to the bottom surface of the above-mentioned tank; and a controller that controls the rotation of the above-mentioned rotating table, the above-mentioned pipe is configured so that one end opening of the above-mentioned pipe contacts the inner wall of the above-mentioned reservoir, and the other end opening of the above-mentioned pipe is separated from the inner wall of the above-mentioned tank, and the above-mentioned controller controls in a manner of performing initial stirring, and the above-mentioned initial stirring comprises: generating an initial inertial flow of the above-mentioned cell suspension by rotating the above-mentioned rotating table in a forward direction around the above-mentioned rotation axis for a first time; and then generating a countercurrent by rotating the above-mentioned rotating table in the opposite direction of the above-mentioned prescribed direction for a second time.

[0013] Another embodiment of the present invention is a dispensing preparation method, which utilizes a dispensing preparation device to stir the cell suspension in the above-mentioned tank, and the above-mentioned dispensing preparation device comprises: a liquid reservoir, which stores the cell suspension, and has an opening, and the discharge tip of the pipette that discharges the cell suspension to the cell storage part repeatedly enters and exits from the above-mentioned opening; a tank, which stores the above-mentioned cell suspension; a piping, which connects the above-mentioned tank and the above-mentioned liquid reservoir, and causes the cell suspension to flow from the above-mentioned tank to the above-mentioned liquid reservoir internally through a pump unit; and a rotating table, which rotates the above-mentioned tank around a rotation axis perpendicular to the bottom surface of the above-mentioned tank. In this dispensing preparation method, there is a step of performing initial stirring, and the above-mentioned initial stirring includes: generating an initial inertial flow of the above-mentioned cell suspension by rotating the above-mentioned rotating table in a forward direction around the above-mentioned rotation axis for a first time; and then generating a countercurrent by rotating the above-mentioned rotating table in the opposite direction of the above-mentioned prescribed direction for a second time.

[0014] Effects of the Invention

[0015] According to the present invention, the suspension can be managed and dispensed without damaging the cells and without causing any unevenness in the cells uniformly contained in the dispensed suspension. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The external appearance structure of the dispensing preparation device as an embodiment of the present invention is shown.

[0017] Figure 2 A functional block diagram of a dispensing preparation device and a dispensing device as an embodiment of the present invention is shown.

[0018] Figure 3 The external appearance structure of a dispensing device as an embodiment of the present invention is shown.

[0019] Figure 4 A schematic cross section showing the relationship between the tip of a pipette and a reservoir in the dispensing preparation device as an embodiment of the present invention.

[0020] Figure 5 A schematic cross section showing the relationship between the tip of a pipette and the suspension surface inside a well in the dispensing preparation device as an embodiment of the present invention. DETAILED DESCRIPTION

[0021] (Embodiment of the present invention)

[0022] Reference Figures 1 to 5 The dispensing preparation device 1 and the dispensing device 10 according to the embodiment of the present invention will be described. Figure 1 The external appearance of the structure of the dispensing preparation device 1 is shown. Figure 2 A functional block diagram of the dispensing preparation device 1 and the dispensing device 10 is shown. Figure 3 The external appearance structure of the dispensing device 10 is shown. Figure 4 The relationship between the tip of the pipette and the suspension surface inside the reservoir 4 of the dispensing preparation device 1 is shown. Figure 5 The schematic diagram shows the relationship between the tip of the pipette and the suspension surface inside the well in the dispensing preparation device 1 .

[0023] [Example 1]

[0024] (Dispensing preparation device and dispensing device)

[0025] The dispensing preparation device 1 includes a bottle tank (hereinafter referred to as a "tank") 2, a rotating table 22, a liquid reservoir 4, a piping system 5, and a controller 6. The dispensing preparation device 1 can be combined so as to constitute a part of the dispensing device 10. The dispensing device 10 can be configured to include, for example, a dispensing head 7 and a plate exchanger 8 in addition to the dispensing preparation device 1.

[0026] The dispensing head 7 causes a pipette having a pipette tip mounted on the front end to move back relative to the interior of the reservoir 4, and stores the cell suspension inside. Then, the cell suspension is injected into the cell storage portion of the container having the cell storage portion. The container having the cell storage portion is, for example, a well plate, and the cell storage portion contains each well. The dispensing head 7 uses the pipette to inject the cell suspension into the well.

[0027] When the dispensing head 7 sucks the cell suspension from the reservoir 4, the pipette is inserted into the reservoir 4. At this time, the tip of the pipette tip reaches a predetermined depth H of at least 1 / 3, preferably 1 / 2 of the depth H from the surface of the cell suspension in the reservoir 4 in the direction of the depth H of the cell suspension. LAfter that, the aspiration of the cell suspension begins. After the aspiration of the specified amount of the cell suspension is completed, the pipette is withdrawn from the interior of the reservoir 4. When the depth H of the cell suspension in the reservoir 4 reaches a specified critical depth S, the dispensing head 7 does not enter or withdraw the pipette from the interior of the reservoir 4. The critical depth S can be freely set within a range where the tip of the pipette tip does not contact the bottom of the reservoir 4, taking into account the manufacturing tolerances of the pipette tip and the pipette, the accuracy of the moving distance of the dispensing head, the deviation of the cell amount in the residual liquid of the cell suspension in the reservoir, and the like.

[0028] When the pipette injects the cell suspension into the cell storage part, the pipette tip 7 does not inject the cell suspension while the pipette tip enters the cell storage part. After the pipette tip completely enters the cell storage part, the pipette tip 7 starts to withdraw from the cell storage part. After the pipette starts to withdraw from the cell storage part, the pipette injects the cell suspension into the cell storage part.

[0029] The plate switch 8 moves the container in the cell storage section into which the cell suspension has not been injected to a predetermined position. Therefore, the dispensing head 7 injects the cell suspension into the cell storage section using a pipette. The plate switch 8 recovers the container in which the cell suspension has been injected into the cell storage section.

[0030] (Dispensing preparation device)

[0031] The liquid reservoir 4 has an opening 4a and contains the cell suspension L R The discharge tip of the pipette that discharges the cell suspension into the cell storage unit repeatedly enters and exits from the opening 4a.

[0032] The tank 2 is arranged next to the liquid storage container 4. The tank 2 includes: a bottle container 21 having an opening on one side and being hollow inside; and a cap 21a capable of closing the opening of the bottle container 21. The bottle container 21 has, for example, a bottom surface 2a, and a cylindrical wall portion extending in the vertical direction along a tank axis 2b passing through the center of the bottom surface 2a and extending perpendicularly to the bottom surface 2a. A hollow portion is defined inside by the cylindrical wall portion and the bottom surface 2a. The cell suspension L is stored inside the tank 2. T . The center of the bottom surface 2a of the bottle container 21 of the tank 2 is, for example, the center of gravity. The tank 2 is typically a cylindrical shape with a circular bottom surface 2a, and the tank axis 2b is an axis passing through the center of the bottom surface 2a. The bottom surface 2a of the tank 2 is not limited to a circle, and may be a polygon headed by a quadrilateral, an ellipse, or other oblong shape. In this case, the main body also has a hollow shape extending in the direction of the tank axis 2b perpendicular to the bottom surface 2a. In this specification, the tank 2 is described as a hollow cylinder with a circular bottom surface 2a. The tank 2 is capable of accommodating a cell suspension in which a specified amount of cells are mixed into a specified culture medium in the hollow portion inside it.

[0033] The piping system 5 connects the tank 2 and the above-mentioned reservoir 4. The pump unit 3 is connected to the piping system 5. The pump unit 3 is, for example, a reciprocating pump having an air cylinder inside. The piping system 5 includes: a tank piping 51 having one end 51a connected to the tank 2; a reservoir piping 52 having one end 52a connected to the reservoir 4; a pump piping 53 having one end 53a connected to the pump unit 3; and a valve 54. The valve 54 is a three-way valve, and the other end 51b of the tank piping 51, the other end 52b of the reservoir piping 52, and the other end 53b of the pump piping 53 are connected to the valve 54. The valve 54 can be, for example, a solenoid valve or a rotary valve. The valve 54 can be switched between a first state and a second state, wherein the first state is to close the connection with the reservoir piping 52 and connect the tank piping 51 to the pump piping 53, and the second state is to close the connection with the tank piping 51 and connect the reservoir piping 52 to the pump piping 53.

[0034] Furthermore, when the valve 54 is switched to the first state, if the pump unit 3 withdraws the cylinder and suctions the inside, the pump pipe 53 and the tank pipe 51 become negative pressure, and the cell suspension in the tank 2 flows into the pump pipe 53 and the tank pipe 51. Next, the valve 54 is switched to the second state, and if the pump unit 3 advances the cylinder to increase the internal pressure, the cell suspension in the pump pipe 53 is sent to the reservoir pipe 52, and finally the cell suspension in the pump pipe 53 is sent to the reservoir 4. Thus, the cell suspension is sent from the tank 2 to the reservoir 4 via the tank pipe 51 and the reservoir pipe 52. The tank pipe 51, the pump pipe 53, and the reservoir pipe 52 suction the cell suspension in the tank 2, and the pump pipe 53 and the reservoir pipe 52 are filled with the cell suspension. In this example, the valve 54 sets the piping system to the first state in conjunction with the retreat of the cylinder of the pump unit 3 , and sets the piping system to the second state in conjunction with the advancement of the cylinder of the pump unit 3 .

[0035] One end 52a of the reservoir pipe 52, which is one end of the pipe system 5, is disposed inside the reservoir 4 so as to be in contact with the inner wall of the reservoir 4. In addition, one end 51a of the tank pipe 51, which is the other end of the pipe system 5, is separated from the inner wall of the tank 2 and is located in the cell suspension L of the cell suspension. TThe tank pipe 51 is fixed at the upper part of the opening of the tank 2, and one end 51a of the tank pipe 51 is inserted into the inside of the tank 2 from the opening of the tank 2. The liquid reservoir 4 has a liquid level detection sensor (not shown) for detecting the liquid level, which can detect the liquid level of the cell suspension inside the liquid reservoir 4. The liquid level detection sensor (not shown) is connected to the controller 6. When the liquid level detection sensor (not shown) detects the liquid level of the liquid reservoir 4, the liquid level is notified to the controller 6. When the controller 6 determines that the liquid level of the liquid level detection sensor is lower than the specified height, it gives an action instruction to the pump unit 3 to make the pump unit 3 operate. The pump unit 3 that receives the action instruction from the controller 6 supplies the cell suspension from the tank 2 to the liquid reservoir 4 through the piping system 5 as described above. The liquid level detection sensor (not shown) can be set to a method other than the method of directly detecting the liquid level. For example, the liquid level can also be indirectly detected based on the calculation of the amount of cell suspension delivered to the liquid reservoir 4. That is, as described above, the amount of the cell suspension delivered by the pump unit 3 corresponds to the decrease in the amount of liquid in the tank 2 and the increase in the amount of liquid in the reservoir 4. As in this embodiment, when the pump unit 3 is a reciprocating pump, in particular, the amount of the cell suspension delivered by the pump unit 3 directly corresponds to the volume of the cylinder of the pump unit 3.

[0036] The dispensing preparation device 1 has a rotating table 22 having a substantially horizontal surface, namely, a loading surface 22a, which is rotated by a motor 23. The rotating table 22 rotates around an axis 23b extending along a vertical rotation axis 23a extending perpendicularly to the bottom surface 2a of the tank 2. The motor 23 rotates the vertical rotation axis 23a. The vertical rotation axis 23a is the rotation center of the motor, and the tank 2 can be placed on the loading surface 22a of the rotating table 22 in a manner centered on the tank axis 2b, which is an axis eccentric to the vertical rotation axis 23a by a distance of a rotation radius d. The rotating table 22 can rotate the tank 2 placed on the loading surface 22a of the rotating table 22 around the vertical rotation axis 23a. The tank 2 placed on the loading surface 22a of the rotating table 22 rotates in a predetermined direction by the rotation of the vertical rotation axis 23a of the rotating table 22. The rotation of the rotating table 22 is a rotation defined under predetermined rotation conditions such as a predetermined rotation radius d, a predetermined angular velocity w, and a predetermined angular acceleration. The tank pipe 51 is inserted into the tank 2, and even if the tank 2 is rotated by the rotating table 22, the position of one end 51a of the tank pipe 51 remains unchanged without twisting. For example, the following structure can be adopted: the cover 21a of the tank 2 is provided with a hole for inserting the tank pipe 51, and the tank pipe 51 is inserted into the inside of the tank 2 through the hole, and the cover 21a holds the tank pipe 51. As a result, the tank pipe 51 will not twist, especially even when the rotation radius of the rotating table 22 is large, and even if the tank 2 is rotated by the rotating table 22, the position of one end 51a of the tank pipe 51 will not change, and no load caused by twisting is applied to the tank pipe 51. The tank pipe 51 can also be configured as a tank pipe 51 that is divided into two parts inside and outside the tank 2 at the cover part, and the inside and outside of the tank 2 are connected by the cover.

[0037] At this time, the rotation radius d of the tank 2 is defined as the distance between the tank axis 2b of the tank 2 and the vertical rotation axis 23a of the turntable 22. For example, the rotation radius d of the tank 2 can be less than the diameter of the inscribed circle of the bottom surface 2a of the tank 2. When the bottom surface 2a of the tank 2 is a perfect circle, it is preferred that the rotation radius d of the tank 2 is less than the diameter of the bottom surface 2a of the tank 2. In addition, when the bottom surface 2a of the tank 2 is a perfect circle, it is further preferred that the rotation radius d of the tank 2 is 1.2 times the radius of the bottom surface 2a of the tank 2. However, as long as the tank 2 can rotate around a rotation axis perpendicular to the bottom surface of the tank 2, the rotation radius d of the tank 2 is not limited thereto and can be freely set.

[0038] The controller 6 controls the rotation of the rotating table 22. When the tank 2 is placed on the mounting surface 22a of the rotating table 22, the controller 6 rotates the rotating table 22, and the initial stirring of the cells in the cell suspension is performed by the rotation of the tank 2. The initial stirring is composed of the initial inertial flow generation (forward rotation) and the countercurrent generation (reverse rotation) performed after the initial inertial flow generation (forward rotation). In the case of only the initial inertial flow generation (forward rotation), the cells only swirl with the swirling flow of the cell suspension in the tank 2, and no stirring effect is produced. Therefore, the initial stirring is first effective by the initial inertial flow generation (forward rotation) and the countercurrent generation (reverse rotation) performed after the initial inertial flow generation (forward rotation). That is, the controller 6 controls so that the rotating table 22 generates the initial inertial flow around the vertical rotation axis 23a in the specified direction of the vertical rotation axis 23a, and then rotates around the vertical rotation axis 23a in the opposite direction of the direction of the initial inertial flow generation. In this specification, the direction of initial inertial flow generation is defined as forward rotation. The direction of rotation of initial inertial flow generation (forward rotation) can be arbitrarily set according to the angular velocity of rotation of the turntable 22. That is, the controller 6 combines initial inertial flow generation (forward rotation) and initial stirring (reverse rotation) to rotate the turntable 22. The initial stirring (reverse rotation) acts as a flow to resist the initial inertial flow generation (forward rotation) generated in the tank 2 to produce a stirring effect in the cell suspension. That is, the initial stirring (reverse rotation) after the initial inertial flow generation (forward rotation) is opposite to the rotation direction of the initial inertial flow generation (forward rotation). The initial inertial flow generation (forward rotation) is performed for the first time, and the countercurrent generation (reverse rotation) is performed for the second time. The second time is less than the first time, and representatively, the second time is less than the first time.

[0039] The angular velocity w of the rotating table 22 based on the controller 6 is defined as an angular velocity within a range that produces a stirring effect on the cells in the cell suspension and does not cause damage to the cells. If the angular velocity of rotation is too slow, the stirring effect on the cells in the cell suspension is reduced. On the other hand, if the angular velocity of rotation is too fast, the cells in the cell suspension may be damaged due to the stirring. The prescribed angular velocity w is, for example, about 2 revolutions per second (2 rps).

[0040] In addition, in the countercurrent generation (reverse rotation) of the initial stirring, in addition to the stirring effect in the cell suspension, there is also an effect of eliminating the rolling of the cell suspension caused by the initial inertial flow generation (forward rotation). That is, the countercurrent generation (reverse rotation) is a flow in the opposite direction of the initial inertial flow generation (forward rotation). Therefore, when a vortex-shaped rolling is generated in the cell suspension in the tank 2 by the initial inertial flow generation (forward rotation), the rolling of the cell suspension generated inside the tank 2 can be eliminated by the countercurrent generation (reverse rotation), thereby preventing the cell suspension from fluctuating inside the tank 2.

[0041] For example, as the initial stirring, the controller 6 first performs 7 turns of initial inertial flow generation (forward rotation) of the turntable 22, and then performs 5 turns of countercurrent generation (reverse rotation) of the turntable 22. For example, representatively, as the initial inertial flow generation (forward rotation), the turntable 22 is rotated 7 to 10 turns, and as the countercurrent generation (reverse rotation), the turntable 22 is rotated 5 to 7 turns. Through the 7 to 10 turns of initial inertial flow generation (forward rotation) and the 5 to 7 turns of countercurrent generation (reverse rotation), the stirring effect and the rolling elimination effect in the three directions of the radial direction, the circumferential direction and the depth direction of the tank 2 can be achieved for the cell suspension in the tank 2. The former stirring effect is an effect for uniformizing the cell distribution in the cell suspension, and the latter rolling elimination effect is an effect for preventing cell damage caused by the flow of the cell suspension caused by stirring in addition to the former stirring effect.

[0042] If the rotating table 22 is rotated 7 to 10 times (the first rotation) as the initial inertial flow generation (forward rotation), a roll-up is generated in the cell suspension, but the roll-up can be eliminated by performing 5 to 7 turns (the second rotation) as the countercurrent generation (reverse rotation). In eliminating the roll-up, it is preferred to rotate the rotating table 22 at a number less than the number of rotations that generate the roll-up flow, particularly at a rotation speed of about 70%. As described above, the rotation (the second rotation) of the countercurrent generation (reverse rotation) plays a role in eliminating the roll-up, so the countercurrent generation (reverse rotation) is less than the rotation (the first rotation) of the initial inertial flow generation (forward rotation). In addition, it is particularly preferred that the rotation (the first rotation) is less than the rotation (the first rotation) of the initial inertial flow generation (forward rotation). For example, the countercurrent generation (reverse rotation) is preferably about 70% of the rotation of the initial inertial flow generation (forward rotation).

[0043] Furthermore, after the initial stirring, at least one additional stirring can be performed. The number of additional stirring times can be freely set according to the distribution state of the cells in the cell suspension. One additional stirring is composed of a combination of additional inertial flow generation (forward) and additional countercurrent generation (reverse) of the turntable 22. That is, one additional stirring is composed of the additional inertial flow generation (forward rotation) of the turntable 22 added after the countercurrent generation (reverse rotation) of the initial stirring and the additional countercurrent generation (reverse rotation) after the additional inertial flow generation (forward rotation). The number of rotations of the additional inertial flow generation (forward rotation) and the number of rotations of the additional countercurrent generation (reverse rotation) can be freely set according to the state of the cells in the tank 2 such as the distribution state of the cells in the cell suspension. In addition, multiple additional stirrings can also be performed. In this case, after the additional countercurrent generation (reverse rotation) in the previous additional stirring, the additional stirring consisting of the additional forward rotation of the turntable 22 and the additional reverse rotation after the additional forward rotation is further performed. Usually, the number of rotations of the additional countercurrent generation (reverse rotation) is less than the number of rotations of the additional inertial flow generation (forward rotation). For example, in one additional stirring, the number of rotations of the additional countercurrent generation (reverse rotation) is 2 times, and the number of rotations of the additional inertial flow generation (forward rotation) is 2 times or 1 time. The additional countercurrent generation (reverse rotation) helps to eliminate the effect of the inertia of the flow that is the cause of the additional inertial flow generation (forward rotation) causing the rolling, and helps the stirring effect of the cells in the suspension.

[0044] Additional stirring, for example, can be performed multiple times. For example, in the example of performing two additional stirrings, the first additional stirring and the second additional stirring after the first additional stirring are performed. In the first additional stirring, the first additional inertial flow generation (forward) and the first additional countercurrent generation (reverse) after the first additional inertial flow generation (forward) are performed. In the second additional stirring, the second additional inertial flow generation (forward) after the first additional countercurrent generation (reverse) and the second additional countercurrent generation (reverse) after the second additional inertial flow generation (forward) are performed. In the first additional stirring and the second additional stirring, the number of rotations of each additional countercurrent generation (reverse rotation) is 2 times, and the number of rotations of the additional inertial flow generation (forward rotation) can be freely set as needed.

[0045] For additional stirring, for example, in one additional stirring, the number of rotations for additional countercurrent generation (reverse rotation) is set to 2 times, and the number of rotations for additional inertial flow generation (forward rotation) is set to 2 times, and such additional stirring can be performed 5 times. That is, for example, after the initial stirring of rotating the turntable 22 by 7 to 10 turns as initial inertial flow generation (forward rotation) and by 5 to 7 turns as initial stirring (reverse rotation), one additional stirring consisting of 2 additional forward rotations and 2 additional reverse rotations is performed continuously 5 times. As a result, inside the tank 2, in the three directions of the radius direction, the circumferential direction, and the depth direction of the tank 2, the cells become uniform in the cell suspension.

[0046] When the cell suspension is stirred in order to make the cells contained in the cell suspension uniform in the cell suspension, if the stirring is performed based on rotation in only one direction, only the inertial flow of the cell suspension is generated, and the cells are rotated together with the cell suspension, and the cells contained in the cell suspension cannot be made uniform in the cell suspension. Therefore, as described above, in either the initial stirring or the additional stirring, after the inertial flow is generated based on the rotation of the forward rotation, the reverse rotation is performed to generate the countercurrent.

[0047] In addition, when the swirling flow of the cell suspension is generated by stirring, a vortex is generated in the cell suspension. If a vortex is generated in the cell suspension, then after stirring ends, inside the cell suspension, cells gather in the center of tank 2 because of the vortex. The additional stirring of the appropriate number of times has the effect of eliminating the vortex generated by the stirring flow. Therefore, in the additional stirring, the number of times and the rotating speed are preferably set to gradually reduce. That is, each rotation in the additional inertial flow generation (forward rotation) and the additional countercurrent generation (reverse rotation) constituting the additional stirring is set to be less than each rotation in the initial inertial flow generation and the countercurrent generation constituting the initial stirring.

[0048] This application claims priority from Japanese Patent Application No. 2022-111714 filed on July 12, 2022, the contents of which are incorporated herein by reference as a part of this application.

[0049] Explanation of symbols

[0050] 1-point injection preparation device

[0051] 2 cans

[0052] 3 pump units

[0053] 4 Liquid reservoir

[0054] 5. Piping system

[0055] 6 Controller

[0056] 7 cents note

[0057] 8-board switch

[0058] 10 dispensing device

[0059] 21 Bottle Container

[0060] 22 Rotating table

[0061] 23 Motor

[0062] 51 tank piping

[0063] 52 Liquid reservoir piping

[0064] 53 Pump piping

[0065] 54 valve

Claims

1. A dispensing preparation device, comprising: A liquid reservoir for storing a cell suspension, the liquid reservoir having an opening, and a discharge tip of a pipette for discharging the cell suspension into the cell storage portion repeatedly entering and exiting the opening; a tank containing the cell suspension; a pipe connecting the tank and the reservoir, and allowing the cell suspension to flow from the tank to the reservoir via a pump unit; a rotating table for rotating the tank about a rotation axis perpendicular to the bottom surface of the tank; and a controller that controls the rotation of the rotating stage, The pipe is arranged such that one end opening of the pipe contacts the inner wall of the liquid storage container and the other end opening of the pipe is separated from the inner wall of the tank. The controller controls in a manner to perform initial stirring, and the initial stirring includes: generating an initial inertial flow of the cell suspension by causing the rotating table to rotate forwardly for a first time in a specified direction around the rotating axis; and then generating a countercurrent by causing the rotating table to rotate reversely for a second time in a direction opposite to the specified direction.

2. The dispensing preparation device according to claim 1, wherein: The center of the bottom surface of the tank is separated from the rotation axis of the rotating table.

3. The dispensing preparation device according to claim 1, wherein: The controller sets the second number of times to be less than or equal to the first number of times.

4. A dispensing preparation device, comprising: The dispensing preparation device according to claim 1; A dispensing head that moves the pipette forward and backward to inject the cell suspension into the cell storage portion of the container having the cell storage portion; and A plate switch moves the container into which the cell suspension has not been injected into the cell storage section to a predetermined position, and recovers the container into which the cell suspension has been injected into the cell storage section.

5. The dispensing preparation device according to claim 4, wherein: During the advancing and retreating of the pipette, after the tip of the pipette reaches a predetermined depth from the surface of the cell suspension in the reservoir in the depth direction of the cell suspension, the dispensing head starts aspirating the cell suspension.

6. The dispensing preparation device according to any one of claims 1 to 5, wherein: The controller performs additional stirring after the initial stirring, and the additional stirring includes: additional inertial flow generation to rotate the turntable in a forward rotation in the specified direction; and additional countercurrent generation to rotate the turntable in a reverse rotation in the opposite direction of the specified direction after the additional inertial flow generation.

7. The dispensing preparation device according to claim 6, wherein: The number of rotations of each rotation in the additional stirring is smaller than the number of rotations of each rotation in the initial stirring.

8. A method for preparing for dispensing, comprising stirring the cell suspension in the tank using a dispensing preparation device, wherein the dispensing preparation device comprises: A liquid reservoir for storing a cell suspension, the liquid reservoir having an opening, through which a discharge tip of a pipette for discharging the cell suspension into the cell storage portion repeatedly enters and exits; a tank containing the cell suspension; a pipe connecting the tank and the reservoir, and allowing the cell suspension to flow from the tank to the reservoir via a pump unit; and a rotating table for rotating the tank about a rotation axis perpendicular to the bottom surface of the tank, The dispensing preparation method includes a step of performing initial stirring, wherein the initial stirring includes: The initial inertial flow of the cell suspension is generated by rotating the rotating table in a predetermined direction around the rotating axis for a first time in a positive direction; and then generating a reverse flow by rotating the rotating table in the opposite direction to the predetermined direction for a second time.

9. The dispensing preparation method according to claim 8, wherein: The second number of times is less than or equal to the first number of times.

10. The dispensing preparation method according to claim 8 or 9, wherein: After the initial stirring, there is a process of performing additional stirring, and the additional stirring includes: additional inertial flow generation, which causes the turntable to rotate in a forward rotation in the specified direction; and additional countercurrent generation, which causes the turntable to rotate in a reverse rotation in the opposite direction of the specified direction after the additional inertial flow is generated.

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