Cell aggregate recovery system

Abstract

Provided is a classification system capable of classifying cell aggregates having a desired size from a culture container in a short time while maintaining a sealed state. The classification system comprises: a culture unit in which a fluid containing cell aggregates is accommodated; and a classification unit that classifies the cell aggregates into first cell aggregates having diameters smaller than a predetermined diameter and second cell aggregates having diameters equal to or greater than a predetermined diameter. The classification unit includes either an extension part in which the fluid flows along the direction of extension and which can be moved in an inclined state in which at least a part thereof is inclined with respect to the vertical direction, or an extension part that is in a fixed inclined state.

Description

Cell Aggregate Recovery System

[0001] The present invention relates to a classification system that can collect cell aggregates while classifying them.

[0002] There is a method in which a suction tube is used to maintain the sealed state and remove the cell aggregate from the culture vessel through the suction tube (see, for example, Patent Document 1).

[0003] International Publication No. 2023 / 013485

[0004] However, when using an aspiration tube, different sedimentation rates are used depending on the size of the cell aggregate. Therefore, the flow rate that can be aspirated through the aspiration tube is determined depending on the size of the cell aggregate, and removal from the culture vessel requires time. Furthermore, depending on the time required for removal, productivity and cell quality may be reduced.

[0005] The present invention has been made in view of the above points, and an object of the present invention is to provide a classification system that can classify cell aggregates of a desired size from a culture vessel in a short period of time while maintaining a sealed state.

[0006] The classification system according to the present invention is characterized by comprising: a culture section in which a fluid containing cell aggregates is accommodated; and a classification section which classifies the cell aggregates into first cell aggregates having a diameter less than a predetermined diameter and second cell aggregates having a diameter equal to or greater than the predetermined diameter, the classification section including either an extension section in which the fluid flows along the extension direction and at least a portion of which can be movable in an inclined state inclined relative to the vertical direction, or an extension section which is fixedly inclined.

[0007] While maintaining the sealed state, cell aggregates having a desired size can be sorted from the culture vessel in a short period of time.

[0008] FIG. 1 is a schematic diagram showing the configuration of a collection system 100 according to a first embodiment. FIG. 2 is a schematic diagram showing the movement of cell aggregates inside a suction tube nozzle 130-1. FIG. 3 is a table showing the relationship between the inclination of the suction tube nozzle 130-1, the circulation flow rate, the collection amount, and the collection rate. FIG. 4 is a schematic diagram showing the configuration of a collection system 200 according to a second embodiment. FIG. 5 is a schematic diagram showing the configuration of a collection system 300 according to a third embodiment. FIG. 6 is a schematic diagram showing the configuration of a collection system 400 according to a fourth embodiment. FIG. 7 is a schematic diagram showing the configuration of a collection system 500 according to a fifth embodiment. FIG. 8 is a schematic diagram showing the configuration of a collection system 600 according to a sixth embodiment. FIG. 9 is a schematic diagram showing the configuration of a collection system 700 according to a seventh embodiment.

[0009] <<<<<Outline of the Present Embodiment>>>>> By including the classification system described below, a recovery system can be constructed that can recover cell aggregates while classifying them.

[0010] <<First Feature>> According to the first feature, there is provided a classification system comprising: a culture section (such as the culture vessel 110 described below) that accommodates a fluid containing cell aggregates; and a classification section (such as the suction tube nozzles 130-1, 130-2, 130-4, and 130-5 described below) that classifies the cell aggregates into first cell aggregates having a diameter less than a predetermined diameter and second cell aggregates having a diameter equal to or greater than the predetermined diameter, the classifying section including either an extension section that allows the fluid to flow along the extension direction and at least a portion of which is movable in an inclined state with respect to the vertical direction, or an extension section that is fixedly inclined.

[0011] The classification system includes a culture unit and a classification unit.

[0012] <Culture section> The culture section contains a fluid containing cell aggregates. The liquid may be a cell suspension containing a culture medium and cell aggregates. Even if it is different from the culture medium and the cell aggregates, it is included in the liquid as long as it is related to the culture of the cell aggregates.

[0013] The culture unit may include a culture tank and an agitator for agitating the liquid.

[0014] The classifying unit classifies the cell aggregates into first cell aggregates and second cell aggregates. The first cell aggregates have a diameter less than a predetermined diameter. The second cell aggregates have a diameter equal to or greater than the predetermined diameter. The classifying unit sorts the cell aggregates based on the predetermined diameter.

[0015] The classification section includes an extension section. Fluid flows through the extension section along the extension direction. The extension section may be selectively tilted or may be fixedly tilted. An extension section that can be selectively tilted can movably transition to either an inclined state or a non-tilted state. The extension section can transition to an inclined state or a non-tilted state by driving a driving member or the like. The non-tilted state is, for example, a vertical state. Therefore, an extension section that can be selectively tilted may be in an inclined state even if it is temporarily in a non-tilted state.

[0016] The extension portion in the fixed inclined state maintains a constant inclined state without displacement. Note that the extension portion may be capable of transitioning to either a first inclined state or a second inclined state, rather than a non-inclined state. The extension portion in the fixed inclined state may be capable of transitioning to a non-inclined state.

[0017] The classification unit can be tilted, so that the flow of the liquid within the classification unit can be adjusted by tilting the classification unit, and the time required for classification into the first cell aggregate and the second cell aggregate can be changed according to the flow of the liquid.

[0018] Furthermore, by including the classification system according to the first feature, it is possible to construct a recovery system that can change the time required for recovery while classifying cell aggregates.

[0019] <<Second Feature>> The second feature is that, in the first feature, the classification section has an intake opening (e.g., intake opening 134-1 described later) that draws in the fluid from the culture section, and an exhaust opening (e.g., exhaust opening 136-1 described later) that is spaced apart from the intake opening along the extension direction of the classification section and that discharges the first cell aggregate.

[0020] The classification section has an intake opening and an exhaust opening, and the intake opening draws fluid from the culture section into the extension section.

[0021] The discharge opening is located apart from the suction opening along the extension direction of the classification section, and the first cell aggregates are discharged from the discharge opening.

[0022] The classifying unit discharges the first cell aggregate from the discharge opening and retains the second cell aggregate in the extension, among the cell aggregates contained in the fluid drawn in from the intake opening. In this way, the cell aggregates can be classified based on the desired diameter.

[0023] Furthermore, by including the classification system according to the second feature, it is possible to construct a recovery system that can recover the first cell aggregates discharged from the discharge opening.

[0024] <<Third Feature>> The third feature is the first or second feature, wherein at least a part of the second cell aggregate moves back and forth along the extending direction of the inclined extending portion of the classifying unit.

[0025] Since the second cell aggregates are moved regularly in the extension portion, the first cell aggregates can be guided toward the discharge opening and discharged from the discharge opening, allowing the cell aggregates to be accurately classified.

[0026] Furthermore, by including the classification system according to the third feature, a recovery system can be constructed that can recover the first cell aggregates that have been accurately classified.

[0027] <<Fourth Feature>> The fourth feature is the first to third features, wherein in the inclined extension portion of the classification section, a first flow region (e.g., first flow region FF described below) in which the second cell agglomerates flow toward the intake opening by reciprocating movement, and a second flow region (e.g., second flow region SF described below) in which the first cell agglomerates flow toward the discharge opening due to the second cell agglomerates flowing through the first flow region are formed.

[0028] The inclined extension of the classification section forms a first flow region and a second flow region. The first flow region is a region where the second cell aggregates flow toward the intake opening by reciprocating. The second flow region is a region where the first cell aggregates flow toward the discharge opening due to the second cell aggregates flowing through the first flow region.

[0029] Furthermore, by including the classification system according to the fourth feature, a recovery system can be constructed that can quickly recover classified first cell aggregates by forming a first flow region and a second flow region.

[0030] <<Fifth Feature>> The fifth feature is the first to fourth features, in which the classifying section includes a straight pipe section (for example, an extension section 132-1 described later) that extends in a constant direction.

[0031] Since it is formed as a straight tube, the cell aggregates can be moved smoothly.

[0032] <<Sixth Feature>> The sixth feature is the first to fifth features, wherein the angle of at least the straight pipe portion with respect to the vertical direction is variable between a first angle (for example, an inclination angle θ1 described later) and a second angle different from the first angle (for example, an inclination angle 0 described later).

[0033] By storing cell aggregates in the straight pipe section when the first angle is set, the recovery time is shortened, and by discharging and recovering the cell aggregates when the second angle is set, recovery accuracy can be maintained.

[0034] <<Seventh Feature>> The seventh feature is the first to sixth features, wherein the classification section has a bent section (for example, bent section 130-2c described later) through which the fluid can flow, and has a first classification section (for example, inclined section 130-2a described later) and a second classification section (for example, vertical section 130-2b described later) connected via the bent section and having different extension directions, and either the first classification section or the second classification section is an inclined extension section of the classification section.

[0035] In the culture section, the device can be arranged so as not to interfere with other components such as a stirring device, and can be arranged so as not to hinder the flow of liquid caused by stirring.

[0036] <<Eighth Feature>> The eighth feature is the first to seventh features, wherein the classification section has a small diameter section (such as suction tube nozzle 130-1 or suction tube nozzle 130-2 described later) having a first inner diameter and a large diameter section (such as expanded diameter section 470 described later) having a second inner diameter larger than the first inner diameter, the small diameter section having the suction opening, and the large diameter section having the discharge opening.

[0037] The small diameter portion allows more cell aggregates to accumulate, while the large diameter portion allows for accurate classification.

[0038] <<<<<Details of the Present Embodiment>>>>> In the manufacturing process of cell aggregates used in cell therapy, separation and concentration are carried out after expansion and culture. For separation and concentration, it is necessary to remove the cell aggregates that have been expanded and cultured in the culture vessel from the culture vessel. If it takes a long time to remove the cell aggregates from the culture vessel, productivity will decrease and there will be concerns about the impact on cell quality (such as adhesion between cell aggregates and reduced activity). Furthermore, the required size range of cell aggregates used in cell therapy varies depending on the cell type and the content of the target treatment. For this reason, it is necessary to separate (classify) cell aggregates having the desired size according to the purpose. The purpose of the recovery system according to the present embodiment is to efficiently remove cell aggregates from the culture vessel while classifying them, while maintaining the sealed state.

[0039] <<<Cells>>> Cells are the subject of culture using collection systems 100, 200, 300, 400, 500, 600, and 700. Culturing cells causes the cells to divide and increase the number of cells. Cells come into contact with each other, forming cell aggregates (cell aggregates). Collection systems 100, 200, 300, 400, 500, 600, and 700 are systems that grow cells and form cell aggregates.

[0040] The cells are not particularly limited as long as they form cell aggregates in a culture medium. The cells are preferably derived from mammals, and preferably from species commonly used in research, such as humans, primates (e.g., monkeys), or mice. Examples of cells include cells used in research on regenerative medicine and cells used as cell preparations. Specific examples include pluripotent stem cells (ES cells and iPS cells), various progenitor cells (e.g., nephron progenitor cells, ureteric bud cells, and interstitial progenitor cells), and various stem cells (e.g., mesenchymal stem cells, neural stem cells, and adipose stem cells). The cells may be cells derived from pluripotent stem cells, immortalized cells, or established cell lines, or primary culture cells isolated from tissue. Depending on the purpose, the cells may be normal cells, cells with a disease, genetic abnormality, or transgene.

[0041] <<<Culture Medium>>> The culture media used in the recovery systems 100, 200, 300, 400, 500, 600, and 700 include new culture media and recycled culture media. New culture media are culture media that are newly supplied to the culture vessel 110 (reactor), which will be described later. Recycling culture media are culture media that have already been supplied to the culture vessel 110 and used for culture, and that have been recycled for circulation or disposal.

[0042] The medium can be selected and used appropriately depending on the cells, and is not particularly limited. Conventional materials and additives useful for cell culture can also be used as appropriate. Examples of basal media include DMEM, DMEMHG, EMEM, IMDM (Iscove's Modified Dulbecco's Medium), GMEM (Glasgow's MEM), RPMI-1640, α-MEM, Ham's Medium F-12, Ham's Medium F-10, Ham's Medium F12K, etc. Additives such as amino acids, vitamins, inorganic salts, proteins (growth factors), glucose, antibiotics, signal transduction inhibitors, reducing agents, and buffers may be added. Serum (preferably mammalian serum, e.g., fetal bovine serum, human serum, etc.) may also be added. Supplements can also be used as a substitute for serum.

[0043] <<<Cell Suspension>>> A cell suspension is a system in which cells or cell aggregates are dispersed in a medium. The culture vessel 110 stores the cells or cell aggregates and the medium as the cell suspension.

[0044] <<<<Agitation>>> Agitation means displacing the cells, cell aggregates, and culture medium in a cell suspension. Agitation displaces the cells and cell aggregates along with the culture medium. The cells and cell aggregates are mainly displaced due to the displacement of the culture medium (convection). Note that the action on the cell suspension can be anything other than agitation as long as it can displace the cells, and can also be an action such as shaking.

[0045] <<<Diameter of Cell Aggregates>>> The diameter of a cell aggregate changes depending on the growth of the cell aggregate. The cell aggregate can be imaged using an imaging device such as a camera, and the diameter of the cell aggregate can be determined from the image results. Furthermore, a correspondence between the time elapsed since the start of culturing the cell aggregate and the diameter of the cell aggregate can be determined in advance, for example, through preliminary experiments, and stored in a memory device (e.g., ROM, RAM, etc.) of the control device. By referring to the correspondence, the diameter of the cell aggregate corresponding to the time since the actual start of culturing can be determined. In this embodiment, it is sufficient to be able to determine the diameter of the cell aggregate at any timing during the culturing process.

[0046] The diameter of a cell aggregate may be any value that indicates the degree of growth (size) of the cell aggregate, and may be any value that indicates the approximate range or degree of the growth process, such as the radius, density distribution, occupied area, or mass of the cell aggregate. For example, diameter may be used appropriately depending on the various laws and equations to be referenced. Furthermore, an easily measurable value may be appropriately selected as the degree of growth of the cell aggregate. In the following, for simplicity, when there is no particular need to distinguish between them, simply the term "diameter" will be used.

[0047] Hereinafter, embodiments will be described with reference to the drawings. <<<Directions, Conditions, etc.>>> <Horizontal Direction> The horizontal direction refers to a direction perpendicular to the gravity of the Earth.

[0048] <Vertical direction> The vertical direction refers to the direction of gravity. It is the direction indicated by the string that suspends an object. It refers to the direction perpendicular to the horizontal direction. Specifically, it refers to the direction in which the central axis CO of the culture vessel 110, the rotation axis RO of the stirring blade 124a, and the central axis AO of the aspirating tube nozzle 130-1 (Figures 1, 2A, 4, 5, 6, 7, etc.) are vertical.

[0049] <Inclined direction> The inclined direction refers to a state inclined from the vertical or horizontal direction, but is a direction that is neither vertical nor horizontal. Specifically, it refers to a direction in which the central axis CO of the culture vessel 110, the rotation axis RO of the stirring blade 124a, or the central axis AO of the aspirating tube nozzle 130-1 is inclined at a predetermined angle from the vertical or horizontal direction.

[0050] For convenience, the angle between the vertical direction and the inclined direction may also be referred to as the inclination angle. The inclination angle refers to the angle between the vertical direction and the reference direction. An acute inclination angle is preferable. For example, the inclination angle is preferably in the range of 5 to 30 degrees. For example, the inclination angle is more preferably in the range of 10 to 20 degrees. The inclination angle is not limited to these ranges, and a suitable range can be determined as appropriate depending on the circulating flow rate of the culture medium, the type and size of the cell aggregates, the desired treatment time, etc.

[0051] <Axial direction> This refers to the direction along the central axis CO of the culture vessel 110 (described below), the rotation axis RO of the stirring blade 124a, or the central axis AO of the aspirating tube nozzle 130-1. The axial direction can be defined mainly for elongated shapes, cylindrical shapes, etc. The axial direction is not limited to a direction along a straight line. It may be any direction along the central axis CO, rotation axis RO, or central axis AO.

[0052] <Vertical State> The vertical state refers to a state in which the direction of the central axis CO of the culture vessel 110, the direction of the rotation axis RO of the agitator blades 124a, and the direction of the central axis AO of the aspirator tube nozzle 130-1 are parallel to the vertical direction. In other words, the vertical state refers to a state in which the central axis CO of the culture vessel 110, the rotation axis RO of the agitator blades 124a, and the central axis AO of the aspirator tube nozzle 130-1 extend along the vertical direction.

[0053] <Tilt State> The tilt state refers to a state in which the direction of the central axis CO of the culture vessel 110, the rotation axis RO of the agitator blades 124a, or the central axis AO of the aspirating tube nozzle 130-1 is tilted. A state in which the central axis CO of the culture vessel 110, the rotation axis RO of the agitator blades 124a, or the central axis AO of the aspirating tube nozzle 130-1 is tilted with respect to the vertical direction is called a tilt state.

[0054] <<<<First embodiment>>>> Fig. 1 is a schematic diagram showing the configuration of a recovery system 100 according to a first embodiment. As shown in Fig. 1, the recovery system 100 is fixedly disposed in an inclined state at a certain inclination angle.

[0055] <<<<Main Components of Recovery System 100 >>> The recovery system 100 mainly includes a culture vessel 110, an agitator 120, and an aspirating tube nozzle 130-1.

[0056] <<Culture Vessel 110>> The recovery system 100 has a culture vessel 110. During the cell culture process, a medium and cells (cell suspension) are contained in the culture vessel 110. The cells are cultured in the culture vessel 110 and grow as cell aggregates.

[0057] The culture vessel 110 has a substantially cylindrical shape. The culture vessel 110 has a substantially cylindrical side wall 112 and a substantially circular bottom 114. The shape of the culture vessel 110 is not limited to a cylindrical shape, and it may have any shape that allows the cell suspension to be stirred by rotating the stirring blades 124a (described later), allowing the cells to be smoothly cultured, and allows the cultured cell aggregates to be smoothly recovered.

[0058] <Central Axis CO> The culture vessel 110 has a central axis CO that extends along the center of the side wall portion 112.

[0059] <<Agitator 120 >> <Configuration of Agitator 120> The agitator 120 agitates the cell suspension stored in the culture vessel 110. The agitator 120 has a drive unit 122, an agitator 124, and a holding member 126.

[0060] The driving unit 122 may be, for example, a magnetic stirrer. The driving unit 122 has a motor (not shown). When the motor rotates, the driving unit 122 uses magnetic force to rotate the stirrer 124, which is disposed at a distance from the driving unit 122. The rotation speed of the motor may be determined appropriately depending on the amount of cell suspension, the number and size of cell aggregates, etc.

[0061] The rotation speed and direction of the motor are controlled by a control device (not shown). The control device has a processor (such as a CPU (Central Processing Unit)), ROM (Read Only Memory), RAM (Random Access Memory), an input / output interface, etc. The ROM, RAM, etc. store programs for controlling the rotation speed and direction of the motor. The processor reads and executes the programs for controlling the rotation speed and direction of the motor from the ROM, RAM, etc.

[0062] It is also possible to rotate the stirring bar 124 by directly connecting it to the shaft of a motor (not shown) without using a magnetic stirrer.

[0063] <Agitation by agitator 120> The cell suspension is agitated by rotation of agitator 124. By agitating the cell suspension, it is possible to prevent the cell aggregates from coming into contact with each other and adhering to each other, or the cell aggregates from coming into contact with side wall 112 or substantially circular bottom 114 of culture vessel 110, throughout the period during which the cell aggregates are collected.

[0064] <Stirring bar 124> The stirring bar 124 has a stirring blade 124a and an extension portion 124b. The stirring blade 124a has a thin plate-like shape that is approximately an isosceles triangle. The stirring blade 124a is located inside the culture vessel 110 and is separated from the drive unit 122. The bottom of the stirring blade 124a faces the bottom 114 of the culture vessel 110.

[0065] The shape of the stirring blades 124a is not limited to a substantially isosceles triangle, and may be any shape that can stir the cell suspension and rotate around the central axis CO. The shape of the stirring blades 124a may be any shape that can form a desired flow of the cell suspension.

[0066] The extension portion 124b has a generally rod-like shape. The extension portion 124b extends from the top of the agitator blade 124a (the top facing the bottom of the agitator blade 124a) in a direction away from the agitator blade 124a. The direction in which the extension portion 124b extends is the direction in which the rotation axis RO of the agitator bar 124 extends. It is preferable to position the agitator bar 124 so that the rotation axis RO roughly coincides with the central axis CO of the culture vessel 110. This positioning allows the cell suspension to be agitated throughout every corner of the culture vessel 110. Note that it is sufficient that the extension direction of the rotation axis RO and the extension direction of the central axis CO are parallel.

[0067] <Holding member 126> The holding member 126 rotatably holds the stirring bar 124. The holding member 126 rotatably holds the stirring bar 124 even if the rotation axis RO of the stirring bar 124 is tilted. By holding the stirring bar 124 with the holding member 126, the rotation axis RO and the central axis CO can be roughly aligned.

[0068] Although an example has been shown in which the rotation axis RO is roughly aligned with the central axis CO, the rotation axis RO may be appropriately determined depending on the inclination angle, the amount of cell suspension, the number and size of cell aggregates, and the bias of the cell aggregates within the culture vessel 110.

[0069] <<Aspiration tube nozzle 130-1>> The aspiration tube nozzle 130-1 is used to sort (classify) cell aggregates among cultured cell aggregates based on a desired size, diameter, etc. Cell aggregates cultured in the culture vessel 110 come in a variety of sizes. The aspiration tube nozzle 130-1 is used to sort cell aggregates based on a desired size, diameter, etc. The classification operation of the aspiration tube nozzle 130-1 will be described later. The aspiration tube nozzle 130-1 is formed from glass, resin, etc., and has a fixed shape.

[0070] <Extended portion 132-1, suction opening 134-1, and discharge opening 136-1> The suction tube nozzle 130-1 has an extended portion 132-1, a suction opening 134-1, and a discharge opening 136-1.

[0071] <Extending portion 132-1> The extending portion 132-1 has an elongated shape. That is, the aspirating tube nozzle 130-1 has an elongated shape. The extending portion 132-1 extends linearly along the longitudinal direction. The extending portion 132-1 extends in an oblique direction. That is, the aspirating tube nozzle 130-1 extends in an oblique direction. In other words, the central axis AO of the aspirating tube nozzle 130-1 extends in an oblique direction.

[0072] As shown in FIG. 2, the extension portion 132-1 has a tubular shape. The extension portion 132-1 has an outer peripheral portion 132O-1 and an inner peripheral portion 132I-1. The outer peripheral portion 132O-1 forms the outer surface of the tube. The inner peripheral portion 132I-1 forms the inner surface of the tube. The outer peripheral portion 132O-1 and the inner peripheral portion 132I-1 are concentrically positioned. A long surrounding region SR, surrounded by the inner peripheral portion 132I-1 and extending along the longitudinal direction, functions as a hollow conduit. The culture medium and cell aggregates can flow through the surrounding region SR. Note that in FIG. 2, the surrounding region SR is shown as a region surrounded by a dashed rectangular line along the inner peripheral portion 132I-1. The extension portion 132-1 has a constant cross-sectional area A1. The cross-sectional area A1 is the area of ​​the inner diameter portion when the aspirating tube nozzle 130-1 is cut along a direction perpendicular to the central axis AO of the aspirating tube nozzle 130-1.

[0073] <Intake Opening 134-1 and Discharge Opening 136-1> The suction tube nozzle 130-1 has two ends in the longitudinal direction. The suction tube nozzle 130-1 has an intake opening 134-1 at one end and an exhaust opening 136-1 at the other end. The extension portion 132-1 extends between the intake opening 134-1 and the exhaust opening 136-1.

[0074] The intake opening 134-1 has a substantially circular opening. The discharge opening 136-1 has a substantially circular opening. The intake opening 134-1, the discharge opening 136-1 and the surrounding region SR are connected to each other. The culture medium and cell aggregates stored in the culture vessel 110 are sucked in through the intake opening 134-1 and guided to the surrounding region SR. Some of the culture medium and cell aggregates guided to the surrounding region SR are discharged through the discharge opening 136-1. The culture medium and cell aggregates stored in the culture vessel 110 are sucked in through the intake opening 134-1 and flow through the surrounding region SR, and some of the cell aggregates are discharged from the discharge opening 136-1 together with the culture medium.

[0075] The suction opening 134-1 and the discharge opening 136-1 have a constant opening area A1 that is the same as the cross-sectional area A1 of the extension portion 132-1. The suction tube nozzle 130-1 also has a cross-sectional area A1. The movement of the cell aggregate guided into the enclosed region SR will be described later.

[0076] <<<<Other Configurations of the Collection System 100 >>> The collection system 100 also includes a collection container 140, a pump 150, a pipe 160a, a pipe 160b, and a pipe 160c.

[0077] The recovery system 100 forms a circulation circuit that circulates the culture medium from the culture vessel 110 using a pump 150. The culture medium is circulated by driving the pump 150. From the culture vessel 110 to the recovery vessel 140, cell aggregates can move together with the culture medium, except for those that remain in the suction tube nozzle 130-1. From the recovery vessel 140 to the culture vessel 110, the culture medium can move.

[0078] <Collection Container 140> The collection container 140 contains cell aggregates that have been classified by the suction tube nozzle 130-1 and discharged from the suction tube nozzle 130-1, together with the culture medium, as a cell suspension. By storing the classified cell aggregates in the collection container 140, the cell aggregates can be collected. The collection container 140 has a substantially cylindrical shape with a constant diameter (cross-sectional area). The collection container 140 has a constant cross-sectional area A2. The axial direction of the collection container 140 is arranged along the vertical direction. The cross-sectional area A2 is the area that occupies the inner diameter portion when the collection container 140 is cut along the direction perpendicular to the central axis (not shown) of the collection container 140.

[0079] The diameter (cross-sectional area A1) of the suction tube nozzle 130-1 is different from the diameter (cross-sectional area A2) of the collection container 140. Specifically, the cross-sectional area A1 of the suction tube nozzle 130-1 is smaller than the cross-sectional area A2 of the collection container 140. Due to the difference in cross-sectional area, the circulation flow rate Q1 generated in the suction tube nozzle 130-1 by driving the pump 150 can be made different from the flow rate Q2 generated in the collection container 140.

[0080] The collection container 140 has an opening at its upper end. The opening of the collection container 140 is sealed by a lid 142 having a sealing member. The lid 142 has two through holes for inserting and attaching tubes 160a and 160b (described below). The two through holes and the tubes 160a and 160b are also sealed by a sealing member or the like. The collection container 140 is hermetically sealed by the lid 142.

[0081] <Pump 150> The pump 150 may be any pump as long as it is capable of adjusting the flow rate. The pump 150 may be, for example, a tube pump, a reciprocating pump, a centrifugal pump, or a propeller pump. The pump 150 has a motor (not shown). The pump 150 can adjust the flow rate by adjusting the rotation speed of the motor, the frequency at which the motor is driven, or the like. Hereinafter, for simplicity, driving the motor of the pump 150 may also be simply referred to as driving the pump 150.

[0082] The rotation speed of the motor, the timing at which the motor is driven, and other parameters are controlled by a control device (not shown). The control device includes a processor (such as a CPU (Central Processing Unit)), a ROM (Read Only Memory), a RAM (Random Access Memory), and an input / output interface. The ROM and RAM store programs for controlling the rotation speed and timing at which the motor is driven. The processor reads and executes the programs for controlling the rotation speed and timing at which the motor is driven from the ROM and RAM.

[0083] The control device that controls the motor of the pump 150 may be the same as or different from the control device that controls the motor of the drive unit 122 of the agitator 120 described above.

[0084] <Driving pump 150> Driving pump 150 allows cell aggregates to be discharged together with the culture medium from culture vessel 110. Furthermore, driving pump 150 allows cell aggregates of a desired size to be guided to and stored in collection vessel 140. Furthermore, driving pump 150 allows the culture medium discharged from collection vessel 140 to be returned to culture vessel 110. The flow rate of culture medium per unit time generated by driving pump 150 is referred to as the circulation flow rate.

[0085] <Tubes 160a, 160b, and 160c> Tubes 160a, 160b, and 160c are made of flexible resin or the like. For simplicity, FIG. 1 illustrates tubes 160a, 160b, and 160c as linear. Tubes 160a, 160b, and 160c have elongated shapes. Tube 160a has a first end 162a and a second end 164a in the longitudinal direction. Tube 160b has a first end 162b and a second end 164b in the longitudinal direction. Tube 160c has a first end 162c and a second end 164c in the longitudinal direction.

[0086] The cross-sectional areas of tubes 160a, 160b, and 160c are not particularly limited. These cross-sectional areas may be appropriately selected so as to form a circulation flow rate and smoothly guide the cell aggregates classified by suction tube nozzle 130-1 into collection container 140. The cross-sectional areas of tubes 160a, 160b, and 160c are the areas that occupy the inner diameter portions when tubes 160a, 160b, and 160c are cut along a direction perpendicular to the longitudinal direction of tubes 160a, 160b, and 160c.

[0087] <Tube 160a> A first end 162a of tube 160a is connected to discharge opening 136-1 of suction tube nozzle 130-1. A second end 164a of tube 160a is fixed so as to be positioned at a predetermined position inside collection container 140. The enclosed region SR of suction tube nozzle 130-1 and tube 160a are in communication with each other. The suction tube nozzle 130-1 is connected to collection container 140 via tube 160a. The culture medium and cell aggregates stored in the culture vessel 110 are classified by the suction tube nozzle 130-1, and the classified cell aggregates, together with the culture medium, are guided to collection container 140 via tube 160a.

[0088] The cross-sectional area of ​​tube 160a is smaller than the cross-sectional area A1 of suction tube nozzle 130-1. The flow rate in suction tube nozzle 130-1 can be made smaller than the flow rate in tube 160a while maintaining the circulation flow rate. By appropriately determining the ratio between the cross-sectional area of ​​tube 160a and the cross-sectional area A1 of suction tube nozzle 130-1, the flow rate in suction tube nozzle 130-1 can be controlled and adjusted so that cell aggregates of the desired size are extracted from culture vessel 110. Furthermore, cell aggregates can be prevented from accumulating in the region including the portion where discharge opening 136-1 of suction tube nozzle 130-1 and first end 162a of tube 160a are connected.

[0089] <Tube 160b> First end 162b of tube 160b is fixed so as to be positioned at a predetermined position inside collection container 140. The height of first end 162b of tube 160b in collection container 140 is higher than the first end 162a of tube 160a. This makes it difficult for cell aggregates contained in collection container 140 to be discharged from tube 160b. Furthermore, tube 160b is positioned at a fixed position relative to collection container 140 so that the height of first end 162b of tube 160b is lower than the liquid level of the cell suspension contained in collection container 140. Only the culture medium is guided into tube 160b.

[0090] A second end 164b of the tube 160b is connected to the first end 152 of the pump 150. The tube 160b is in communication with the pump 150. The collection container 140 is connected to the pump 150 via the tube 160b. The second end 164a of the tube 160a is fixed so as to be positioned at a predetermined position inside the collection container 140.

[0091] <Tube 160c> A first end 162c of the tube 160c is connected to the second end 154 of the pump 150. A second end 164c of the tube 160c is fixed so as to be disposed at a predetermined position inside the culture vessel 110. The tube 160c and the pump 150 are in communication with each other.

[0092] <<<Circulation Circuit>>> Cell aggregates, except for those remaining in the suction tube nozzle 130-1, are moved together with the medium from the culture vessel 110 to the collection vessel 140. The collection vessel 140 contains the cell suspension.

[0093] As described above, collection container 140 is sealed by lid 142 or the like. Furthermore, tube 160b communicates with pump 150, and tube 160c communicates with pump 150. Tubes 160b and 160c communicate with each other via pump 150. Furthermore, the height of first end 162b of tube 160b in collection container 140 is higher than second end 164a of tube 160a and lower than the liquid level of the cell suspension contained in collection container 140.

[0094] With this configuration, the pressure inside collection container 140 is reduced by driving pump 150, and cell aggregates, together with the culture medium, are guided from culture container 110 to collection container 140 via suction tube nozzle 130-1. Furthermore, by driving pump 150, only the culture medium of the cell suspension guided to and contained in collection container 140 can be aspirated from collection container 140. The culture medium aspirated from collection container 140 returns to culture container 110 via tube 160b, pump 150, and tube 160c. Note that the cell aggregates of the cell suspension contained in collection container 140 remain in collection container 140 and can be collected.

[0095] A circulation circuit through which the culture medium can circulate can be formed by the culture vessel 110, the agitator 120, the suction tube nozzle 130-1, the collection vessel 140, the pump 150, the pipe 160a, the pipe 160b, and the pipe 160c. By driving the pump 150, the culture medium can circulate through the circulation circuit at a circulation flow rate.

[0096] <<<<Movement of Cell Aggregates Within the Aspirate Tube Nozzle 130-1>>> Figure 2B is a cross-sectional view showing an outline of the movement of cell aggregates within the aspirate tube nozzle 130-1. In Figure 2B, multiple white circular regions represent cell aggregates.

[0097] <Top Generator UG and Bottom Generator BG> The suction tube nozzle 130-1 has an extension 132-1, which has a tubular shape. The outer periphery 132O-1 of the extension 132-1 has multiple generators along the longitudinal direction. Generators refer to the straight lines at each position when a curved surface (tubular (cylindrical) surface) is drawn (formed) by moving a straight line. By tilting the suction tube nozzle 130-1, the cross section of the suction tube nozzle 130-1 can be defined as having an uppermost generator UG and a lowermost generator BG. The cross section of the suction tube nozzle 130-1 is a plane formed by cutting the suction tube nozzle 130-1 perpendicular to the central axis AO of the suction tube nozzle 130-1 (see FIG. 2B ).

[0098] <In the case of a vertical state> When the suction tube nozzle 130-1 is in a vertical state, the culture medium rises in the region formed between the plurality of cell aggregates settling within the suction tube nozzle 130-1. When the suction tube nozzle 130-1 is in a vertical state, the culture medium is sucked through the suction opening 134-1 and then sucked vertically upward within the suction tube nozzle 130-1. This is the direction opposite to the direction of gravity, and the direction of movement of the cell aggregates is determined by the magnitude relationship between the suction force and gravity. In other words, the movement direction of the cell aggregates is either up or down in the vertical direction.

[0099] <In the case of tilted state> On the other hand, as shown in Figures 1 and 2, tilting the suction tube nozzle 130-1 causes cell aggregates attempting to settle to move and gather in the area closer to the bottom bus line BG than in the area closer to the top bus line UG due to the action of gravity. As the gathered cell aggregates move toward the suction opening 134-1, the area closer to the bottom bus line BG becomes more likely to be occupied by cell aggregates. Due to the uneven distribution of cell aggregates along the cross section of the suction tube nozzle 130-1 (see Figure 2B), the culture medium moves and flows in the area closer to the top bus line UG, which is not occupied by cell aggregates, rather than in the area closer to the bottom bus line BG, which is occupied by cell aggregates. Tilting the suction tube nozzle 130-1 causes such movement of the cell aggregates and the culture medium, and two main flow regions, a first flow region FF and a second flow region SF, are formed in the enclosed region SR. In the cross-sectional view shown in Fig. 2B, the dashed, approximately circular shape is an imaginary line showing the boundary BD between the first flow region FF and the second flow region SF. In the cross-sectional view shown in Fig. 2A, the boundary BD is shown as a dashed straight line located between the uppermost bus line UG and the lowermost bus line BG along the longitudinal direction of the suction tube nozzle 130-1. The region outside the boundary BD is the first flow region FF, and the region inside the boundary BD is the second flow region SF.

[0100] <First flow region FF> The first flow region FF is a region where cell aggregates gather and then flow (settle) together with the culture medium toward the suction opening 134-1. That is, the first flow region FF is a region where a flow is formed in which the cell aggregates settle together with the culture medium. In particular, in the first flow region FF, cell aggregates are more likely to settle in the region close to the bottom bus line BG. The cell aggregates settle faster in the region close to the bottom bus line BG and settle more slowly as they move away from the bottom bus line BG.

[0101] The culture medium flows in succession together with the cell aggregates from the suction opening 134-1. Therefore, some of the cell aggregates that have settled near the suction opening 134-1 flow out from the suction opening 134-1, while others move to the second flow region SF and rise again. Whether they flow out from the suction opening 134-1 or rise again is determined appropriately depending on the amount and diameter of the cell aggregates, the flow of the culture medium near the suction opening 134-1, and other factors.

[0102] <Second flow region SF> The second flow region SF is a region where, as the culture medium rises toward the discharge opening 136-1, the cell aggregates also flow toward the discharge opening 136-1. That is, the second flow region SF is a region where a flow of rising cell aggregates is formed. In particular, in the second flow region SF, cell aggregates are more likely to rise in regions close to the uppermost generatrix UG. In other words, the cell aggregates rise faster in regions close to the uppermost generatrix UG and rise more slowly as they move away from the uppermost generatrix UG.

[0103] The cell aggregates that rise to the vicinity of the discharge opening 136-1 may flow out from the discharge opening 136-1 or may move to the first flow region FF and settle again. Whether the cell aggregates flow out from the discharge opening 136-1 or settle again is determined appropriately depending on the amount and diameter of the cell aggregates, the flow of the medium near the discharge opening 136-1, etc.

[0104] <<Circulating Cell Aggregates>> Due to the formation of the first flow region FF, large (heavy) cell aggregates of a predetermined diameter (size, mass) or greater settle toward the intake opening 134-1 and then move to the second flow region SF. Furthermore, due to the formation of the second flow region SF, the cell aggregates that have moved to the second flow region SF rise toward the discharge opening 136-1 and then move again to the first flow region FF. In this way, cell aggregates of a predetermined size or greater flow alternately between the first flow region FF and the second flow region SF and circulate within the suction tube nozzle 130-1. By circulating cell aggregates of a predetermined size or greater within the suction tube nozzle 130-1, they can be retained within the suction tube nozzle 130-1.

[0105] <<Cell aggregates to be discharged>> By forming the second flow region SF, cell aggregates smaller (lighter) than the reference diameter (size, mass) rise toward the discharge opening 136-1 and are discharged from the discharge opening 136-1 together with the culture medium.

[0106] <<Classification of Cell Aggregates>> Cell aggregates equal to or larger than a predetermined reference size remain within the suction tube nozzle 130-1. Meanwhile, cell aggregates smaller than the predetermined size are discharged through the discharge opening 136-1. Thus, by using the inclined suction tube nozzle 130-1, the formation of the first flow region FF and the second flow region SF allows cell aggregates to be classified based on a predetermined diameter (size, mass). Cell aggregates equal to or larger than the predetermined size that remain within the suction tube nozzle 130-1 are discharged through the suction opening 134-1 of the suction tube nozzle 130-1. Meanwhile, cell aggregates smaller than the predetermined size are discharged through the discharge opening 136-1 of the suction tube nozzle 130-1 and collected in the collection container 140.

[0107] <Distribution of cell aggregates in the suction tube nozzle 130-1> Figure 2B is a cross-sectional view showing the distribution of cell aggregates in the suction tube nozzle 130-1 at the II section shown in Figure 2A. In Figure 2B, multiple white circular regions also represent cell aggregates.

[0108] As described above, the area close to the bottom generatrix BG is occupied by the settling cell aggregates. In other words, the cross section of the first flow region FF (the size along the direction perpendicular to the longitudinal direction of the suction tube nozzle 130-1) is occupied by the settling cell aggregates and becomes larger.

[0109] On the other hand, since the area close to the bottom generating line BG is occupied by cell aggregates, the area close to the top generating line UG where the culture medium can flow is reduced. In other words, the cross section of the second flow region SF (the size along the direction perpendicular to the longitudinal direction of the suction tube nozzle 130-1) is reduced by the cell aggregates in the first flow region FF.

[0110] The circulation flow rate of the culture medium discharged from the suction tube nozzle 130-1 is determined by the operation of the pump 150. To maintain this circulation flow rate, the flow rate of the culture medium flowing in the second flow region SF, where the cross section has become smaller, increases, and the movement speed of the cell aggregates also increases. As a result, the flow rate of the cell suspension attempting to rise in the second flow region SF increases. In other words, the flow of the culture medium can promote the rise of the cell aggregates in the second flow region SF. As described above, the formation of the second flow region SF causes cell aggregates that are smaller (lighter) than a reference diameter (size, mass) to be discharged from the discharge opening 136-1. As the culture medium flows, the cell aggregates are also rapidly discharged from the discharge opening 136-1, allowing the cell aggregates to be collected in a short period of time.

[0111] <<<Relationship between the tilt of the suction tube nozzle 130-1 and the circulation flow rate>>> The relationship between the tilt of the suction tube nozzle 130-1 and the circulation flow rate is the relationship between the tilt angle θ of the suction tube nozzle 130-1 and the circulation flow rate obtained under the condition that cell aggregates of a predetermined size or larger are retained in the suction tube nozzle 130-1 and discharged from the suction opening 134-1, and cell aggregates of less than the predetermined size are discharged and recovered from the discharge opening 136-1 of the suction tube nozzle 130-1.

[0112] When the inclination angle θ of the suction tube nozzle 130-1 is set to 0 degrees, even if the circulation flow rate is increased to F0, both cell agglomerates larger than a predetermined size and cell agglomerates smaller than a predetermined size are discharged from the suction opening 134-1 of the suction tube nozzle 130-1.

[0113] When the inclination angle θ of the suction tube nozzle 130-1 is set to θ1, which is greater than 0 degrees, even if the circulation flow rate is increased to F1, which is greater than F0, cell aggregates of a predetermined size or larger will remain in the suction tube nozzle 130-1 and be discharged from the suction opening 134-1, and cell aggregates of a size smaller than the predetermined size will be discharged and recovered from the discharge opening 136-1 of the suction tube nozzle 130-1.

[0114] As the tilt angle θ of the suction tube nozzle 130-1 increases from 0 degrees, the circulation flow rate is expected to monotonically increase and to exist within a range where it exceeds F0. Note that as the tilt angle θ of the suction tube nozzle 130-1 increases from 0 degrees, the circulation flow rate may initially increase, and then become approximately constant or decrease. The circulation flow rate relative to the tilt angle θ of the suction tube nozzle 130-1 is determined depending on the size and type of cell aggregate, the type of culture medium, etc.

[0115] In other words, the circulation flow rate can be increased as the tilt angle θ of the suction tube nozzle 130-1 increases. That is, in order to retain cell aggregates of a predetermined size or larger in the suction tube nozzle 130-1 and discharge them from the suction opening 134-1, and to discharge and recover cell aggregates of a size smaller than the predetermined size from the discharge opening 136-1 of the suction tube nozzle 130-1, the circulation flow rate can be increased by increasing the tilt angle θ of the suction tube nozzle 130-1.

[0116] <<Recovery Rate of Cell Aggregates>> Figure 3 is a table showing the relationship between the inclination of the suction tube nozzle 130-1, the circulation flow rate, the recovery amount, and the recovery rate. Figure 3 shows an example of the relationship obtained under conditions for retaining cell aggregates of a predetermined (desired constant) size or larger in the suction tube nozzle 130-1 and discharging them from the suction opening 134-1, and discharging and recovering cell aggregates smaller than the predetermined (desired constant) size from the discharge opening 136-1 of the suction tube nozzle 130-1.

[0117] The recovery amount indicates the amount of cell aggregates recovered from the culture vessel 110 via the suction tube nozzle 130-1 into the recovery vessel 140. The recovery rate is the proportion of cell aggregates that are smaller than a predetermined (desired constant) size among the cell aggregates recovered into the recovery vessel 140.

[0118] 3, when the tilt angle θ of the suction tube nozzle 130-1 is 0 (vertical state) and the circulation flow rate is F0, the recovery amount is V0 and the recovery rate is R0. When the tilt angle θ of the suction tube nozzle 130-1 is θ1 (inclined state) and the circulation flow rate is F1, the recovery amount is V1 and the recovery rate is R1.

[0119] The relationship of the inclination angle θ of the suction tube nozzle 130-1 is 0<θ1. The relationship of the corresponding circulation flow rate is F0<F1, as shown in Figure 3. In other words, the circulation flow rate can be made larger in an inclined state than in a vertical state.

[0120] Furthermore, as shown in Figure 3, the relationship of the recovered amount was V0 < V1. That is, the recovered amount was greater in the inclined state than in the vertical state.

[0121] 3, the relationship of the recovery rate was C1<C0. The recovery rate was higher in the vertical state than in the inclined state.

[0122] From the above, by tilting the suction tube nozzle 130-1 from a vertical position, it is possible to increase the recovery amount while maintaining a certain level of recovery rate. That is, by tilting the suction tube nozzle 130-1 and increasing the circulation flow rate to F1 by driving the pump 150, cell aggregates of a predetermined size or larger are retained in the suction tube nozzle 130-1 and discharged through the suction opening 134-1, while cell aggregates of a size smaller than the predetermined size are discharged through the discharge opening 136-1 of the suction tube nozzle 130-1, thereby increasing the amount that can be recovered. In other words, by tilting the suction tube nozzle 130-1, it is possible to increase the circulation flow rate and increase the amount of cell aggregates that can be discharged from the suction tube nozzle 130-1, thereby shortening the time required for recovery.

[0123] As described above, the relationship in Figure 3 is determined under the condition that cell aggregates of a predetermined (desired constant) size or larger are retained in the suction tube nozzle 130-1 and discharged through the suction opening 134-1, and cell aggregates of a size smaller than the predetermined (desired constant) size are discharged through the discharge opening 136-1 of the suction tube nozzle 130-1 for recovery. Therefore, if the size of the cell aggregates to be recovered is changed, the tilt angle θ of the suction tube nozzle 130-1, the circulation flow rate, the recovery amount, and the recovery rate will differ. When recovering cell aggregates of various sizes, these conditions can be determined in advance through preliminary experiments, etc., and the tilt angle θ of the suction tube nozzle 130-1 and the circulation flow rate can be changed depending on the size of the cell aggregates to be recovered.

[0124] <<<<Recovery Process of Cell Aggregates by Recovery System 100>>> An outline of the recovery process of cell aggregates by the recovery system 100 will be described. In the following, it is assumed that the cell aggregates are dispersed in the culture vessel 110 by the agitator 120.

[0125] Due to the circulation flow rate Q1 generated by driving pump 150, cell aggregates smaller than a predetermined size in culture vessel 110 are classified by suction tube nozzle 130-1 and discharged together with the culture medium from suction tube nozzle 130-1. The cell aggregates and culture medium discharged from suction tube nozzle 130-1 are introduced into collection vessel 140 via tube 160a.

[0126] The cross-sectional area A2 of the collection container 140 is larger than the cross-sectional area A1 of the suction tube nozzle 130-1 (A1<A2). Therefore, the flow rate Q2 / A2 within the collection container 140 is smaller than the flow rate Q1 / A1 within the suction tube nozzle 130-1. The cell aggregates introduced into the collection container 140 can remain within the collection container 140 due to gravitational settling. Due to the difference in flow rate and the gravitational settling of the cell aggregates, the culture medium can be discharged from the collection container 140 while the cell aggregates remain within the collection container 140.

[0127] The pump 150 can be driven by determining the circulation flow rate Q1 corresponding to the inclination of the suction tube nozzle 130-1 according to the diameter of the cell aggregate to be removed from the culture vessel 110. By driving the pump 150, the culture medium discharged from the collection vessel 140 is returned to the culture vessel 110 via the pipes 160b and 160c.

[0128] The stirring speed and circulation flow rate Q1 may be determined depending on the concentration of the cell aggregates in the culture vessel 110, etc.

[0129] <<<<<Second Embodiment>>>>> Figure 4 is a schematic diagram showing the configuration of a recovery system 200 according to a second embodiment. In Figure 4, the same components as those in the first embodiment are denoted by the same reference numerals. In Figure 4, the recovery container 140, pump 150, pipe 160a, pipe 160b, and pipe 160c are the same as those in the first embodiment and are therefore omitted. As shown in Figure 4, the culture container 110 of the recovery system 200 is fixedly arranged in a vertical state, unlike the recovery system 100. Below, differences from the recovery system 100 will mainly be described.

[0130] <Culture Vessel 110> The configuration and materials of the culture vessel 110 are basically the same as those of the recovery system 100. The arrangement of the culture vessel 110 differs from that of the recovery system 100. The direction of the central axis CO of the culture vessel 110 is vertical, which differs from the arrangement of the culture vessel 110 in the recovery system 100. Therefore, the bottom 114 of the culture vessel 110 extends horizontally.

[0131] <Suction Tube Nozzle 130-2> Unlike the collection system 100, the collection system 200 has a suction tube nozzle 130-2.

[0132] The suction tube nozzle 130-2 has an inclined portion 130-2a, a vertical portion 130-2b, and a bent portion 130-2c. The inclined portion 130-2a, the vertical portion 130-2b, and the bent portion 130-2c give the suction tube nozzle 130-2 an elongated shape overall. The cross-sectional area of ​​the suction tube nozzle 130-2 is the area of ​​the inner diameter when the inclined portion 130-2a and the vertical portion 130-2b are cut along a direction perpendicular to the longitudinal direction of the inclined portion 130-2a and the vertical portion 130-2b.

[0133] <Inclined portion 130-2a> The inclined portion 130-2a is inclined at an angle θ with respect to the vertical direction. The inclined portion 130-2a extends in the inclined direction. The inclined portion 130-2a has an elongated shape. The inclined portion 130-2a extends linearly along the longitudinal direction. The inclined portion 130-2a has a tubular shape. The inclined portion 130-2a has an intake opening 134-1.

[0134] The aspirating tube nozzle 130-2 is positioned so that the suction opening 134-1 of the inclined portion 130-2a faces the sidewall 112 of the culture vessel 110. That is, the direction in which the inclined portion 130-2a extends is the inclination direction, and the direction in which the extension portion 124b of the stirring bar 124 extends is the vertical direction, forming an angle θ with the inclined portion 130-2a. By inclining the inclined portion 130-2a so that the suction opening 134-1 faces the sidewall 112 of the culture vessel 110, the inclined portion 130-2a is less likely to interfere with the stirring blades 124a of the stirring bar 124, allowing the medium and cell aggregates to be sufficiently stirred. Furthermore, by inclining the inclined portion 130-2a, the degree of freedom in the size and shape of the stirring blades 124a of the stirring bar 124 can be increased.

[0135] <Vertical section 130-2b> The vertical section 130-2b extends in the vertical direction. The vertical section 130-2b has an elongated shape. The vertical section 130-2b extends linearly along the longitudinal direction. The vertical section 130-2b has a tubular shape. The vertical section 130-2b has a discharge opening 136-1.

[0136] <Bent portion 130-2c> The inclined portion 130-2a and the vertical portion 130-2b are connected via the bent portion 130-2c. The bent portion 130-2c has a short tubular shape. The bent portion 130-2c connects the inclined portion 130-2a extending in the inclined direction to the vertical portion 130-2b extending in the vertical direction at an angle θ. The inclined portion 130-2a and the vertical portion 130-2b communicate with each other via the bent portion 130-2c. Instead of the bent portion 130-2c, a curved member may be used to connect the inclined portion 130-2a extending in the inclined direction to the vertical portion 130-2b extending in the vertical direction.

[0137] <Inclined portion 130-2a, vertical portion 130-2b, and bent portion 130-2c> The inclined portion 130-2a, vertical portion 130-2b, and bent portion 130-2c have an outer periphery and an inner periphery (not shown), similar to the extension portion 132-1 of the suction tube nozzle 130-1 of the recovery system 100. The outer periphery constitutes the outer surface of the inclined portion 130-2a, vertical portion 130-2b, and bent portion 130-2c. The inner periphery constitutes the inner surface of the inclined portion 130-2a, vertical portion 130-2b, and bent portion 130-2c. The outer periphery and inner periphery are positioned concentrically. A long surrounding region SR, surrounded by the inner periphery and extending along the longitudinal direction, functions as a hollow conduit. The culture medium and cell aggregates can flow through the surrounding region SR.

[0138] The culture medium and cell aggregates flow in the inclined portion 130-2a in the same manner as in the suction tube nozzle 130-1 of the first embodiment. Two flow regions, a first flow region FF and a second flow region SF, are formed in the inclined portion 130-2a. Cell aggregates having a diameter equal to or larger than a predetermined reference diameter remain within the inclined portion 130-2a. On the other hand, cell aggregates having a diameter smaller than the predetermined reference diameter are discharged from the inclined portion 130-2a. In this manner, by using the inclined portion 130-2a, the formation of the first flow region FF and the second flow region SF enables the cell aggregates to be classified based on the predetermined diameter. Only the culture medium and cell aggregates that flow out of the inclined portion 130-2a flow through the vertical portion 130-2b and are discharged from the discharge opening 136-1.

[0139] <Other Shapes of Suction Opening 134-1> Although only an example in which the inclined portion 130-2a is linearly inclined so that the suction opening 134-1 faces the side wall portion 112 of the culture vessel 110 has been shown, the present invention is not limited to this. By appropriately changing the shape of the suction tube nozzle 130-2 according to the size and shape of the stirring blades 124a of the stirrer 124, such as by curving the suction tube nozzle 130-2 in a spiral shape, interference with the stirring blades 124a can be avoided.

[0140] <<<<<Third Embodiment>>>>> Figure 5 is a schematic diagram showing the configuration of a collection system 300 according to a third embodiment. In Figure 5, the same components as in the first embodiment are denoted by the same reference numerals. In Figure 5, the collection container 140, pump 150, pipe 160a, pipe 160b, and pipe 160c are the same as in the first embodiment and are therefore omitted.

[0141] As shown in Fig. 5, unlike the recovery systems 100 and 200, the recovery system 200 is configured to be able to alternate between a vertical state and an inclined state. The recovery system 300 has the same configuration as the recovery system 100, except that it can alternate between a vertical state and an inclined state. In other words, when it is in an inclined state, it is in the same state as the first embodiment. Below, differences from the recovery system 100 will be mainly described.

[0142] The retrieval system 300 includes a movable platform 370 and an actuator (not shown).

[0143] <Movable Platform 370> The movable platform 370 is equipped with the culture vessel 110, the agitator 120, and the aspirating tube nozzle 130-1.

[0144] The culture vessel 110, the agitator 120, and the aspirating tube nozzle 130-1 are arranged to maintain fixed positions relative to each other on the movable base 370. By displacing the movable base 370, the culture vessel 110, the agitator 120, and the aspirating tube nozzle 130-1 can be moved while maintaining their relative positions.

[0145] <Actuator> The actuator is composed of a motor, a solenoid, or the like. The actuator is controlled by a control device (not shown). The actuator is driven by a control signal from the control device to change the tilt of the movable table 370. By driving the actuator, the culture vessel 110, the agitator 120, and the aspirating tube nozzle 130-1 mounted on the movable table 370 can be selectively transitioned to either a vertical state or an inclined state while maintaining their relative positions.

[0146] When the suction tube nozzle 130-1 transitions to the tilted state, the culture medium and cell aggregates flow in the same manner as the suction tube nozzle 130-1 of the first embodiment. Two flow regions, a first flow region FF and a second flow region SF, are formed in the suction tube nozzle 130-1. Cell aggregates that are equal to or larger than a predetermined reference size remain within the suction tube nozzle 130-1. On the other hand, cell aggregates that are smaller than the predetermined size are discharged from the suction tube nozzle 130-1.

[0147] In this manner, by using the suction tube nozzle 130-1 that has transitioned to the inclined state, the cell aggregates can be classified based on a predetermined diameter by forming the first flow region FF and the second flow region SF. Only the culture medium and cell aggregates that flow out from the suction tube nozzle 130-1 are discharged from the discharge opening 136-1.

[0148] <Control process for transition between vertical state and tilted state> First, the processor of the control device (not shown) outputs a control signal to drive the actuator, displacing the movable base 370 and placing the suction tube nozzle 130-1 in a tilted state (tilt angle θ1 in FIG. 3) (step S31). This places the suction tube nozzle 130-1 in the same state as in the first embodiment.

[0149] Next, the processor of the control device outputs a control signal to drive pump 150, setting the circulation flow rate to F1 (FIG. 3) (step S33). As a result, with suction tube nozzle 130-1 in an inclined state, more cell aggregates are stored within suction tube nozzle 130-1 and classified.

[0150] The processor of the control device determines whether a first predetermined time has elapsed since the pump 150 was driven to set the circulation flow rate to F1 (step S35). The first predetermined time is a time determined in advance through a preliminary experiment or the like. For example, the time it takes for the cell aggregates to accumulate in the suction tube nozzle 130-1 may be measured, and the first predetermined time may be determined based on the measurement results.

[0151] When the first predetermined time has elapsed, the processor of the control device outputs a control signal to drive the actuator, displacing the movable base 370 and bringing the suction tube nozzle 130-1 into a vertical position (tilt angle 0 in FIG. 3) (step S37). Furthermore, the processor of the control device outputs a control signal to drive the pump 150, bringing the circulation flow rate to F0 (FIG. 3) (step S39).

[0152] This maintains the state in which the cell aggregates are stored in the suction tube nozzle 130-1, and the cell aggregates stored in the suction tube nozzle 130-1 are collected into the collection container 140 while being classified.

[0153] The processor of the control device determines whether a second predetermined time has elapsed since the pump 150 was driven to set the circulation flow rate to F0 (step S41). The second predetermined time is a time determined in advance through a preliminary experiment or the like.

[0154] When the second predetermined time has elapsed, the processor of the control device returns the process to step S31 again, sets the inclined state (inclination angle θ1), and sets the circulation flow rate to F1. The second predetermined time is the time it takes for the amount of cell aggregates stored in the aspirating tube nozzle 130-1 to decrease.

[0155] As a result, when the amount of cell aggregates stored in the suction tube nozzle 130-1 decreases, the actuator can be driven again to tilt the nozzle, allowing the cell aggregates to be stored in the suction tube nozzle 130-1.

[0156] By repeating the processes of steps S31 to S41, the time required to recover the cell aggregates can be shortened compared to the first embodiment in which the cell aggregates are fixed in an inclined state.

[0157] The processor of the control device terminates the process when a third predetermined time has elapsed. The third predetermined time is a time determined in advance through a preliminary experiment or the like.

[0158] In the above-described process, whether or not to switch control is determined depending on time, but the state inside the suction tube nozzle 130-1 may be detected using a sensor, camera, or the like (not shown), and the process may be switched depending on the detection results. For example, the suction tube nozzle 130-1 may be made of a material that allows the interior to be observed, and a sensor may be used to detect the number of cell aggregates present inside the suction tube nozzle 130-1. Depending on the detection results, the nozzle may be switched to either a vertical state or an inclined state.

[0159] In this way, when collecting cell aggregates into the collection container 140, the suction tube nozzle 130-1 is changed from an inclined state to a vertical state, so that the accuracy of collecting cell aggregates can be made equivalent to that when the nozzle is fixed in a vertical state.

[0160] In the first embodiment, the suction tube nozzle 130-1 can only be inclined, so that an increase in the circulation flow rate can result in a decrease in collection accuracy. In contrast, in the third embodiment, the suction tube nozzle 130-1 can be in either an inclined or vertical state, so that when in the inclined state, cell aggregates are stored within the suction tube nozzle 130-1, shortening the collection time, and when in the vertical state, the cell aggregates are collected in the collection container 140, thereby maintaining collection accuracy.

[0161] <<<<<Fourth Embodiment>>>>> Figure 6 is a schematic diagram showing the configuration of a collection system 400 according to a fourth embodiment. In Figure 6, the same components as in the first embodiment are denoted by the same reference numerals. In Figure 6, the collection container 140, pump 150, pipe 160a, pipe 160b, and pipe 160c are also the same as in the first embodiment and are therefore omitted.

[0162] 6, the recovery system 400 is fixedly disposed in an inclined state at a certain inclination angle, similar to the recovery system 100. The following mainly describes the differences from the recovery system 100.

[0163] The recovery system 400 has a suction tube nozzle 130-4. The suction tube nozzle 130-4 is made of glass, resin, or the like, and has a fixed shape.

[0164] The suction tube nozzle 130-4 includes the suction tube nozzle 130-1 and an expanded diameter portion 470. In other words, the suction tube nozzle 130-4 according to the fourth embodiment has a structure in which the expanded diameter portion 470 is provided at the discharge opening 136-1 of the suction tube nozzle 130-1 of the first embodiment.

[0165] <Expanded diameter portion 470> The expanded diameter portion 470 extends in the vertical direction. The expanded diameter portion 470 has an elongated shape. The expanded diameter portion 470 extends linearly along the longitudinal direction. The expanded diameter portion 470 has a tubular shape. The expanded diameter portion 470 has a discharge opening 472. In addition, the lower end of the expanded diameter portion 470 has an opening that communicates with the discharge opening 136-1.

[0166] <Aspiration tube nozzle 130-1, expanded diameter portion 470, and enclosed region SR> The suction tube nozzle 130-1 and expanded diameter portion 470 have an outer circumferential portion and an inner circumferential portion (not shown), similar to the extension portion 132-1 of the suction tube nozzle 130-1 of the collection system 100. The outer circumferential portion constitutes the outer surface of the suction tube nozzle 130-1 and expanded diameter portion 470. The inner circumferential portion constitutes the inner surface of the suction tube nozzle 130-1 and expanded diameter portion 470. The long enclosed region SR, which is surrounded by the inner circumferential portion and extends along the longitudinal direction, functions as a hollow conduit. Culture medium and cell aggregates can flow through the enclosed region SR.

[0167] <Flow of Culture Medium and Cell Aggregates> As in the first embodiment, the culture medium and cell aggregates flow through the suction tube nozzle 130-1. Two flow regions, a first flow region FF and a second flow region SF, are formed in the suction tube nozzle 130-1. Cell aggregates that are equal to or larger than a predetermined reference size remain within the suction tube nozzle 130-1 and are discharged from the suction opening 134-1. On the other hand, cell aggregates that are smaller than the predetermined size are discharged and collected from the discharge opening 136-1 of the suction tube nozzle 130-1.

[0168] Only the culture medium and cell aggregates discharged from the discharge opening 136-1 of the suction tube nozzle 130-1 flow through the enlarged diameter portion 470 and are discharged from the discharge opening 472 of the enlarged diameter portion 470.

[0169] The expanded diameter section 470 has an inner diameter larger than the inner diameter of the suction tube nozzle 130-1. That is, the expanded diameter section 470 has a cross-sectional area larger than the cross-sectional area of ​​the suction tube nozzle 130-1. By increasing the cross-sectional area of ​​the expanded diameter section 470, the flow rate in the expanded diameter section 470 can be reduced. The cross-sectional area of ​​the expanded diameter section 470 may be determined according to the desired flow rate in the expanded diameter section 470. The cross-sectional area of ​​the expanded diameter section 470 refers to the area of ​​the inner diameter portion when the expanded diameter section 470 is cut in a direction perpendicular to the vertical direction in which the expanded diameter section 470 extends (i.e., horizontally).

[0170] <Inclined Recovery and Vertical Recovery> The suction tube nozzle 130-1 is inclined (for example, at an inclination angle θ1) and the expanded diameter section 470 is vertical (for example, at an inclination angle of 0), so that the suction tube nozzle 130-1 performs inclined recovery and the expanded diameter section 470 performs vertical recovery. The inner diameter (cross-sectional area) of the expanded diameter section 470 is determined by the classification flow rate for vertical recovery of cell aggregates of a desired size. The suction tube nozzle 130-1 can accumulate more cell aggregates, while the expanded diameter section 470 can classify them with high accuracy. If the inner diameter (cross-sectional area) of the suction tube nozzle 130-1 is increased, the amount of cell aggregates sucked in increases, which raises concerns about a deterioration in the agitation state of the culture vessel 110 near the suction opening 134-1.

[0171] However, by increasing the inner diameter (cross-sectional area) of the expanded diameter section 470, it is possible to separate the role of the expanded diameter section 470 from that of the suction tube nozzle 130-1, and the cell aggregates can be collected efficiently.

[0172] <<<<Fifth Embodiment>>>> Figure 7 is a schematic diagram showing the configuration of a collection system 500 according to a fifth embodiment. In Figure 7, the same components as in the first embodiment are denoted by the same reference numerals. In Figure 7, the collection container 140, pump 150, pipe 160a, pipe 160b, and pipe 160c are the same as in the first embodiment and are therefore omitted.

[0173] 7, the incubation vessel 110 of the recovery system 500 is fixedly arranged in a vertical state, similar to the recovery system 200. The following mainly describes the differences from the recovery system 100.

[0174] The recovery system 500 has a suction tube nozzle 130-5. The suction tube nozzle 130-5 is made of glass, resin, or the like, and has a fixed shape.

[0175] The suction tube nozzle 130-5 includes the suction tube nozzle 130-2 and an expanded diameter portion 470. In other words, the suction tube nozzle 130-5 according to the fifth embodiment has a structure in which the expanded diameter portion 470 of the fourth embodiment is provided in the discharge opening 136-1 of the suction tube nozzle 130-2 of the second embodiment.

[0176] Similar to the fourth embodiment, the expanded diameter portion 470 has a discharge opening 472. The lower end of the expanded diameter portion 470 also has an opening, which communicates with the discharge opening 130-2b.

[0177] <Aspiration tube nozzle 130-2, expanded diameter portion 470, and enclosed region SR> The suction tube nozzle 130-2 and expanded diameter portion 470 have an outer circumferential portion and an inner circumferential portion (not shown), similar to the extension portion 132-1 of the suction tube nozzle 130-1 of the collection system 100. The outer circumferential portion constitutes the outer surface of the suction tube nozzle 130-2 and expanded diameter portion 470. The inner circumferential portion constitutes the inner surface of the suction tube nozzle 130-2 and expanded diameter portion 470. The long enclosed region SR, which is surrounded by the inner circumferential portion and extends along the longitudinal direction, functions as a hollow conduit. Culture medium and cell aggregates can flow through the enclosed region SR.

[0178] <Flow of Culture Medium and Cell Aggregates> As in the first embodiment, the culture medium and cell aggregates flow through the suction tube nozzle 130-2. Two flow regions, a first flow region FF and a second flow region SF, are formed in the inclined portion 130-2a of the suction tube nozzle 130-2. Cell aggregates equal to or larger than a predetermined reference size remain within the inclined portion 130-2a and are discharged from the suction opening 134-1. On the other hand, cell aggregates smaller than the predetermined size are discharged and collected from the discharge opening 130-2b of the suction tube nozzle 130-2. Only the culture medium and cell aggregates discharged from the discharge opening 130-2b flow through the enlarged diameter portion 470 and are discharged from the discharge opening 472 of the enlarged diameter portion 470.

[0179] <Inclined Recovery and Vertical Recovery> The expanded diameter section 470 has an inner diameter larger than that of the suction tube nozzle 130-2. That is, the expanded diameter section 470 has a cross-sectional area larger than that of the suction tube nozzle 130-2. Increasing the cross-sectional area of ​​the expanded diameter section 470 can reduce the flow rate through the expanded diameter section 470. Since the suction tube nozzle 130-2 is inclined (e.g., at an inclination angle θ1) and the expanded diameter section 470 is vertical (e.g., at an inclination angle of 0), the suction tube nozzle 130-2 performs inclined recovery, while the expanded diameter section 470 performs vertical recovery. The inner diameter (cross-sectional area) of the expanded diameter section 470 is determined by the classification flow rate for vertical recovery of cell aggregates of the desired size. The suction tube nozzle 130-2 can accumulate more cell aggregates, while the expanded diameter section 470 allows for accurate classification.

[0180] If the inner diameter (cross-sectional area) of the suction tube nozzle 130-2 is increased, the amount of cell aggregates sucked in increases, which raises concerns about a decrease in the state of agitation in the culture vessel 110 near the suction opening 134-1. However, by increasing the inner diameter (cross-sectional area) of the expanded diameter section 470, it is possible to separate the roles of the expanded diameter section 470 and the suction tube nozzle 130-2, thereby enabling efficient recovery of cell aggregates.

[0181] <<<<<Sixth Embodiment>>>>> Fig. 8 is a schematic diagram showing the configuration of a collection system 600 according to a sixth embodiment. In Fig. 8, the same components as those in the first embodiment are denoted by the same reference numerals. In Fig. 8, the collection container 140, pump 150, pipe 160a, pipe 160b, and pipe 160c are the same as those in the first embodiment and are therefore omitted. As shown in Fig. 8, the culture container 110 of the collection system 600 is fixedly arranged in a vertical state, unlike the collection system 100. The following mainly describes the differences from the collection system 100.

[0182] <Culture Vessel 110> The configuration and materials of the culture vessel 110 are basically the same as those of the recovery system 100. The arrangement of the culture vessel 110 differs from that of the recovery system 100. The direction of the central axis CO of the culture vessel 110 is vertical, which differs from the arrangement of the culture vessel 110 in the recovery system 100. Therefore, the bottom 114 of the culture vessel 110 extends horizontally.

[0183] <Agitator 120> The configuration of the agitator 120 is basically the same as that of the recovery system 200 according to the second embodiment. The arrangement of the agitator 120 differs from that of the recovery system 100 according to the first embodiment. The direction of the rotation axis RO of the agitator 124 of the agitator 120 is vertical, which differs from the arrangement of the agitator 124 in the recovery system 100. In other words, the central axis CO of the culture vessel 110 and the rotation axis RO of the agitator 124 are vertical, which differs from that of the recovery system 100.

[0184] <Suction Tube Nozzle 130 - 2 > The configuration and materials of the suction tube nozzle 130 - 2 are basically the same as those of the recovery system 100 .

[0185] Similar to the suction tube nozzle 130-1, the suction tube nozzle 130-2 has an extension portion 132-1, an intake opening 134-1, and an exhaust opening 136-1.

[0186] The extension portion 132-1 has a similar configuration to the suction tube nozzle 130-1. The extension portion 132-1 extends in an inclined direction. The extension portion 132-1 is inclined at an inclination angle θ with respect to the vertical direction. Therefore, the suction tube nozzle 130-1 extends in an inclined direction. In other words, the central axis AO of the suction tube nozzle 130-1 extends in the inclined direction.

[0187] In the first embodiment, the central axis CO of the culture vessel 110, the rotation axis RO of the agitator blades 124a, and the central axis AO of the aspirate tube nozzle 130-1 are arranged along the inclined direction. In contrast, in the sixth embodiment, the central axis CO of the culture vessel 110 and the rotation axis RO of the agitator blades 124a extend vertically, and only the aspirate tube nozzle 130-1 extends in the inclined direction. The aspirate tube nozzle 130-1 is fixedly provided by a holding member or the like (not shown) so that it extends in a fixed inclined direction.

[0188] The aspirating tube nozzle 130-2 is positioned so that the suction opening 134-1 faces the sidewall 112 of the culture vessel 110. That is, the aspirating tube nozzle 130-2 is positioned so that the central axis AO of the aspirating tube nozzle 130-1 moves away from the central axis CO of the culture vessel 110 as it moves downward. By tilting the aspirating tube nozzle 130-2 so that the suction opening 134-1 faces the sidewall 112 of the culture vessel 110, the aspirating tube nozzle 130-2 is less likely to interfere with the stirring blades 124a of the stirring bar 124, allowing the culture medium and cell aggregates to be sufficiently stirred. Furthermore, tilting the aspirating tube nozzle 130-2 allows for greater freedom in the size and shape of the stirring blades 124a of the stirring bar 124.

[0189] The culture medium and cell aggregates flow in the suction tube nozzle 130-2 in the same manner as in the suction tube nozzle 130-1 of the first embodiment. Two flow regions, a first flow region FF and a second flow region SF, are formed in the suction tube nozzle 130-2. Cell aggregates equal to or larger than a predetermined reference size remain within the suction tube nozzle 130-2 and are discharged from the suction opening 134-1. Meanwhile, cell aggregates smaller than the predetermined size are discharged and collected from the discharge opening 136-1 of the suction tube nozzle 130-2. In this way, by using the inclined suction tube nozzle 130-2, the formation of the first flow region FF and the second flow region SF allows the cell aggregates to be classified based on a predetermined diameter.

[0190] <<<<<Seventh Embodiment>>>>> Figure 9 is a schematic diagram showing the configuration of a collection system 700 according to a seventh embodiment. In Figure 9, the same components as those in the first and second embodiments are denoted by the same reference numerals. In Figure 9, the collection container 140, pump 150, pipe 160a, pipe 160b, and pipe 160c are the same as those in the first embodiment and are therefore omitted. As shown in Figure 9, the culture container 110-3 of the collection system 700 is fixedly arranged in a vertical state, similar to the second, fifth, and sixth embodiments. Below, differences from the collection system 100 and the collection system 200 will mainly be described.

[0191] <Incubation Vessel 110-3> The structure and materials of the incubation vessel 110-3 are basically the same as those of the collection system 100. In the collection systems 100 and 200, the incubation vessel 110 and the aspirating tube nozzle 130-1 and 130-2 are configured separately from each other. Therefore, the position and orientation of the aspirating tube nozzle 130-1 and 130-2 can be adjusted relative to the incubation vessel 110. In contrast, the incubation vessel 110-3 is formed integrally with the aspirating tube nozzle 130-3.

[0192] The suction tube nozzle 130-3 has an extension 132-3, an intake opening 134-3, and an exhaust opening 136-3.

[0193] The culture vessel 110-3 has a through-hole formed near the bottom 114. The suction opening 134-3 of the suction tube nozzle 130-3 is fixed so as to communicate with the through-hole formed near the bottom 114 of the culture vessel 110-3. The suction tube nozzle 130-3 forms an inclination angle θ with respect to the central axis CO of the culture vessel 110-3 and extends so as to move away from the culture vessel 110-3 as it extends upward. The central axis AO of the suction tube nozzle 130-3 extends in the inclined direction.

[0194] By forming the aspirating tube nozzle 130-3 integrally with the culture vessel 110-3, a holding member for holding the aspirating tube nozzle 130-3 is not required. Furthermore, since the tilt angle θ of the aspirating tube nozzle 130-3 can be kept constant, adjustment of the tilt angle θ is also not required. Furthermore, since the aspirating tube nozzle 130-3 is disposed outside the culture vessel 110-3, the aspirating tube nozzle 130-3 does not interfere with the stirring blades 124a of the stirring bar 124. This configuration allows for sufficient stirring of the culture medium and cell aggregates, and allows for greater freedom in the size and shape of the stirring blades 124a of the stirring bar 124.

[0195] 9 shows a configuration in which the suction opening 134-3 of the suction tube nozzle 130-3 is formed near the bottom 114 of the culture vessel 110-3, but the position of the suction opening 134-3 of the suction tube nozzle 130-3 is not limited to this. The position of the suction opening 134-3 can be determined appropriately depending on the type and amount of culture medium, the type and distribution of cell aggregates within the culture vessel 110-3, the rotation speed of the stirrer 124, etc. It is sufficient that the suction tube nozzle 130-3 is formed integrally with the culture vessel 110-3.

[0196] The culture medium and cell aggregates flow in the suction tube nozzle 130-3 in the same manner as in the suction tube nozzle 130-1 of the first embodiment. Two flow regions, a first flow region FF and a second flow region SF, are formed in the suction tube nozzle 130-3. Cell aggregates equal to or larger than a predetermined reference size remain within the suction tube nozzle 130-3 and are discharged from the suction opening 134-3. Meanwhile, cell aggregates smaller than the predetermined size are discharged and collected from the discharge opening 136-3 of the suction tube nozzle 130-3. In this way, by using the inclined suction tube nozzle 130-3, the formation of the first flow region FF and the second flow region SF allows the cell aggregates to be classified based on a predetermined diameter.

[0197] <Other Configurations of Culture Vessel 110-3> In the example shown in Figure 9, the aspirating tube nozzle 130-3 is configured to be integral with the culture vessel 110-3. The configuration of the culture vessel 110-3 is not limited to this. The aspirating tube nozzle 130-3 may be configured to be detachable and separate from the culture vessel 110-3.

[0198] For example, a generally cylindrical protruding portion (not shown) for attaching the aspirate tube nozzle 130-3 is provided on the side of the culture vessel 110-3. The aspirate tube nozzle 130-3 can be detachably attached to the culture vessel 110-3 by inserting the end of the aspirate tube nozzle 130-3, which is formed separately, into the protruding portion. By constructing the aspirate tube nozzle 130-3 separately from the culture vessel 110-3, it is possible to use aspirate tube nozzle 130-3 made of different materials from the culture vessel 110-3, or to use an aspirate tube nozzle 130-3 having a desired length or shape.

[0199] Furthermore, the position of the through-hole and protrusion for attaching the aspirating tube nozzle 130-3, the angle relative to the vertical direction, etc. can be set as desired, thereby increasing the degree of freedom in selecting the aspirating tube nozzle 130-3 and the culture vessel 110-3.

[0200] <<<<<Scope of the Embodiments>>>>> As described above, the first to fifth embodiments have been described. However, the descriptions and drawings that form part of this disclosure should not be understood as limiting. Various embodiments not described herein are also included.

[0201] This can be applied to cell culture in which cell aggregates of a desired size need to be classified and recovered from a culture vessel in a short time while maintaining a sealed state. CROSS-REFERENCE TO RELATED APPLICATIONS

[0202] This application claims priority based on Japanese Patent Application No. 2024-018123, filed with the Japan Patent Office on February 8, 2024, the entire disclosure of which is incorporated herein by reference in its entirety.

[0203] 100, 200, 300, 400, 500 Recovery system 110 Culture vessel 130-1, 130-2, 130-4, 130-5 Aspirator nozzle 132-1 Extension part

Claims

1. A classification system comprising: a culture section that contains a fluid containing cell aggregates; and a classification section that classifies the cell aggregates into first cell aggregates having a diameter less than a predetermined diameter and second cell aggregates having a diameter equal to or greater than the predetermined diameter, the classification section including either an extension section in which the fluid flows along an extension direction and at least a portion of which can be moved to an inclined state inclined relative to the vertical direction, or an extension section that is fixedly inclined.

2. The classification system described in claim 1, wherein the classification section has an intake opening for intake of the fluid from the culture section, and an exhaust opening spaced apart from the intake opening along the extension direction of the classification section and for exhausting the first cell aggregate.

3. A classification system as described in claim 2, wherein at least a portion of the second cell aggregate moves back and forth along the extending direction of the inclined extending portion of the classification section.

4. A classification system as described in claim 3, wherein the inclined extension portion of the classification section forms a first flow region in which the second cell agglomerates flow toward the intake opening by reciprocating movement, and a second flow region in which the first cell agglomerates flow toward the discharge opening due to the second cell agglomerates flowing through the first flow region.

5. The classification system according to claim 1, wherein the classification section includes a straight pipe section that extends in a constant direction.

6. The classification system according to claim 5, wherein the angle of at least the straight pipe section relative to the vertical direction can be changed between a first angle and a second angle different from the first angle.

7. A classification system as described in claim 1, wherein the classification unit has a bent portion through which the fluid can flow, and comprises a first classification unit and a second classification unit connected via the bent portion and extending in different directions, and either the first classification unit or the second classification unit is an inclined extension portion of the classification unit.

8. The classification system according to claim 2, wherein the classification section has a small diameter section having a first inner diameter and a large diameter section having a second inner diameter larger than the first inner diameter, the small diameter section having the intake opening, and the large diameter section having the discharge opening.

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