Cell aggregate recovery system

Abstract

To provide 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.SOLUTION: A classification system comprises: a culture unit in which 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 the predetermined diameter, the classification unit including either an extension part in which the fluid flows along a 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.SELECTED DRAWING: Figure 1

Description

[Technical Field]

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

[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). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2023 / 013485 Summary of the Invention [Problem to be solved by the invention]

[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. [Means for solving the problem]

[0006] The classification system according to the present invention is characterized by: a culture section in which a fluid containing cell aggregates is accommodated; The device is equipped with a classification unit that classifies 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, and includes an extension unit that allows the fluid to flow along the extension direction and includes either an extension unit that can be movable in an inclined state with at least a portion inclined relative to the vertical direction, or an extension unit that is fixedly inclined. [Effects of the Invention]

[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. [Brief explanation of the drawings]

[0008] [Figure 1] 1 is a schematic diagram showing the configuration of a recovery system 100 according to a first embodiment. [Figure 2] FIG. 10 is a schematic diagram showing the movement of cell aggregates in the suction tube nozzle 130-1. [Figure 3] 10 is a table showing the relationship between the inclination of the aspirating tube nozzle 130-1, the circulation flow rate, the recovery amount, and the recovery rate. [Figure 4] FIG. 10 is a schematic diagram showing the configuration of a recovery system 200 according to a second embodiment. [Figure 5] FIG. 10 is a schematic diagram showing the configuration of a recovery system 300 according to a third embodiment. [Figure 6] FIG. 10 is a schematic diagram showing the configuration of a recovery system 400 according to a fourth embodiment. [Figure 7] FIG. 10 is a schematic diagram showing the configuration of a recovery system 500 according to a fifth embodiment. [Figure 8] FIG. 10 is a schematic diagram showing the configuration of a recovery system 600 according to a sixth embodiment. [Figure 9] FIG. 12 is a schematic diagram showing the configuration of a recovery system 700 according to a seventh embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0009] <<<<<Outline of this 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, A culture section (for example, a culture vessel 110 described later) in which a fluid containing cell aggregates is accommodated; A classification system is provided which includes a classification unit (e.g., suction tube nozzles 130-1, 130-2, 130-4, 130-5, etc., described below) that classifies 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, and which includes either an extension unit in which the fluid flows along the 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 unit that is fixedly inclined.

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

[0012] <Cultivation Department> 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 the liquid 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 classification 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 classification 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] An extension portion that is in a fixed inclined state maintains a constant inclined state without displacement. However, it may be capable of transitioning to either a first inclined state or a second inclined state, rather than a non-inclined state. An extension portion that is in a fixed inclined state may be in any state as long as it cannot be in a non-inclined state.

[0017] The classification unit can be tilted, so that the flow of liquid within the classification unit can be adjusted by tilting the classification unit. The time required for classification into the first cell aggregate and the second cell aggregate can be changed according to the flow of 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 classifying unit includes: an intake opening (for example, intake opening 134-1, which will be described later) that draws in the fluid from the culture unit; a discharge opening (e.g., discharge opening 136-1, which will be described later) that is spaced apart from the suction opening along the extension direction of the classification unit and through which the first cell aggregate is discharged; It has.

[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>> A third feature is the first or second feature, At least a portion 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 in the extension portion are moved regularly, the first cell aggregates can be guided toward the discharge opening and discharged from the discharge opening, allowing the cell aggregates to be classified accurately.

[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 that in the first to third features, In the inclined extension portion of the classification portion, A first flow region (e.g., a first flow region FF described later) in which the second cell aggregate flows toward the suction opening by reciprocating movement; A second flow region (e.g., a second flow region SF described below) in which the first cell aggregates flow toward the discharge opening due to the second cell aggregates flowing through the first flow region; is 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 the classified first cell aggregates by forming the first flow region and the second flow region.

[0030] <<Fifth feature>> The fifth feature is that in the first to fourth features, The classification section includes a straight pipe section (for example, an extension section 132-1, which will be 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 that in the first to fifth features, At least the straight pipe section has an angle with respect to the vertical direction that can be changed between a first angle (e.g., an inclination angle θ1, which will be described later) and a second angle different from the first angle (e.g., an inclination angle 0, which will be 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] <<7th feature>> The seventh feature is that in the first to sixth features, The classification section has a bent portion (e.g., a bent portion 130-2c described later) through which the fluid can flow, and has a first classification section (e.g., an inclined portion 130-2a described later) and a second classification section (e.g., a vertical portion 130-2b described later) that are connected via the bent portion and extend in different directions from each other, Either the first classifying section or the second classifying section serves as an inclined extension of the classifying 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] <<8th feature>> The eighth feature is the first to seventh features, The classification section has a small diameter section (e.g., a suction tube nozzle 130-1 or a suction tube nozzle 130-2, which will be described later) having a first inner diameter, and a large diameter section (e.g., an expanded diameter section 470, which will be described later) having a second inner diameter larger than the first inner diameter, the small diameter portion has the intake opening; The large diameter portion has 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 this embodiment>>>> In the manufacturing process of cell aggregates used in cell therapy, separation and concentration are performed after expansion and culture. For separation and concentration, the cell aggregates expanded and cultured in the culture vessel must be removed from the culture vessel. If it takes a long time to remove the cell aggregates from the culture vessel, productivity decreases and there are concerns about effects 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 target treatment. Therefore, it is necessary to separate (classify) cell aggregates having the desired size according to the purpose. The recovery system according to this embodiment aims to efficiently remove cell aggregates from the culture vessel while classifying them, while maintaining the sealed state.

[0039] <<<Cell>>> Cells are the subject of cultivation using collection systems 100, 200, 300, 400, 500, 600, and 700. Culturing cells causes them 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 culture medium. Cells are preferably derived from mammals, 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). 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, 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 are classified as new culture media and recycled culture media. New culture media are culture media that are newly supplied to the culture vessel 110 (reactor) described below. Recycling culture media are culture media that have already been supplied to the culture vessel 110 and used for culture, and are recycled for circulation or disposal.

[0042] The medium can be selected and used appropriately depending on the cells, and is not particularly limited. Conventionally known 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, and Ham's Medium F12K. 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. In the culture vessel 110, cells or cell aggregates and a medium are stored as a cell suspension.

[0044] <<<Stirring>>> Agitation means displacing the cells, cell aggregates, and medium in the cell suspension. Agitation displaces the cells and cell aggregates along with the medium. The cells and cell aggregates are mainly displaced due to the displacement of the 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] <<<Cell aggregate diameter>>> The diameter of a cell aggregate changes depending on the growth of the cell aggregate. The cell aggregate is 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 is 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 the 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 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, an embodiment will be described with reference to the drawings. <<<Direction, condition, etc.>>> <Horizontal direction> The horizontal direction is the direction perpendicular to the force of 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 (FIGS. 1, 2A, 4, 5, 6, 7, etc.) are vertical.

[0049] <Tilt 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 preferred. 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> The axial direction 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, the rotation axis RO, or the 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 agitating blades 124a, and the direction of the central axis AO of the aspirating 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 agitating blades 124a, and the central axis AO of the aspirating tube nozzle 130-1 extend along the vertical direction.

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

[0054] <<<<First embodiment>>>> Fig. 1 is a schematic diagram showing the configuration of a recovery system 100 according to the 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 the recovery system 100>>> The recovery system 100 mainly includes a culture vessel 110, a stirring device 120, and a suction tube nozzle 130-1.

[0056] <<Culture vessel 110>> The recovery system 100 includes a culture vessel 110. In the process of culturing cells, a medium and cells (cell suspension) are contained in the culture vessel 110. In the culture vessel 110, the cells are cultured 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] <Center axis CO> The culture vessel 110 has a central axis CO that extends along the center of the sidewall portion 112 .

[0059] <<Agitator 120>> <Configuration of the Stirring Device 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 can be, for example, a magnetic stirrer. The driving unit 122 has a motor (not shown). When the motor rotates, the driving unit 122 rotates a stirrer 124, which is disposed at a distance from the driving unit 122, by magnetic force. 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 the agitator 120> The cell suspension is stirred by the rotation of the stirrer 124. By stirring the cell suspension, it is possible to prevent the cell aggregates from coming into contact with each other and adhering to each other, and to prevent the cell aggregates from coming into contact with the side wall 112 or the approximately circular bottom 114 of the culture vessel 110, throughout the period during which the cell aggregates are collected.

[0064] <Stirrer 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 agitating blades 124a is not limited to a substantially isosceles triangle shape, and may be any shape that can agitate the cell suspension and rotate around the central axis CO. The shape of the agitating 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 impeller 124a (the top facing the bottom of the impeller 124a) in a direction away from the impeller 124a. The direction in which the extension portion 124b extends is the direction in which the rotation axis RO of the stirrer 124 extends. It is preferable to position the stirrer 124 so that the rotation axis RO roughly coincides with the central axis CO of the culture vessel 110. By positioning the stirrer in this way, the cell suspension can be stirred 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 to each other.

[0067] <Retaining member 126> Holding member 126 rotatably holds stirrer 124. Holding member 126 rotatably holds stirrer 124 even if rotation axis RO of stirrer 124 is tilted. By holding stirrer 124 with holding member 126, rotation axis RO and 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] <<Suction tube nozzle 130-1>> The suction tube nozzle 130-1 is used to sort (classify) cell aggregates among the cultured cell aggregates based on a desired size, diameter, etc. The cell aggregates cultured in the culture vessel 110 come in a variety of sizes. The suction tube nozzle 130-1 is used to sort the cell aggregates based on a desired size, diameter, etc. The classification operation of the suction tube nozzle 130-1 will be described later. The suction tube nozzle 130-1 is made of glass, resin, etc., and has a fixed shape.

[0070] <Extension part 132-1, suction opening 134-1, discharge opening 136-1> The suction tube nozzle 130-1 is An extension portion 132-1; an intake opening 134-1; a discharge opening 136-1; It has.

[0071] <Extension part 132-1> The extension portion 132-1 has an elongated shape. That is, the aspirating tube nozzle 130-1 has an elongated shape. The extension portion 132-1 extends linearly along the longitudinal direction. The extension 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 exhaust 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 suction opening 134-1 has a substantially circular opening. The discharge opening 136-1 has a substantially circular opening. The suction opening 134-1, the discharge opening 136-1 and the surrounding area SR are connected to each other. The culture medium and cell aggregates stored in the culture vessel 110 are sucked in through the suction opening 134-1 and guided to the surrounding area SR. Some of the culture medium and cell aggregates guided to the surrounding area 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 suction opening 134-1 and flow through the surrounding area 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 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 Recovery 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 collection 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 collection vessel 140, cell aggregates can move together with the culture medium, except for those remaining at the suction tube nozzle 130-1. From the collection 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 has a processor (such as a CPU (Central Processing Unit)), a ROM (Read Only Memory), a RAM (Random Access Memory), an input / output interface, and other components. The ROM, RAM, and other components 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, RAM, and other components.

[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 the pump 150> By driving pump 150, cell aggregates can be discharged together with the culture medium from culture vessel 110. Furthermore, by driving pump 150, cell aggregates of a desired size can be guided and stored in collection vessel 140. Furthermore, by driving pump 150, the culture medium discharged from collection vessel 140 can 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] <Pipe 160a, pipe 160b, pipe 160c> Tube 160a, tube 160b, and tube 160c are made of flexible resin or the like. For simplicity, tube 160a, tube 160b, and tube 160c are shown in a straight line in FIG. 1. Tube 160a, tube 160b, and tube 160c have an elongated shape. 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 the tube 160a is connected to the discharge opening 136-1 of the suction tube nozzle 130-1. A second end 164a of the tube 160a is fixed so as to be positioned at a predetermined position inside the collection container 140. The enclosed region SR of the suction tube nozzle 130-1 is in communication with the tube 160a. The suction tube nozzle 130-1 is connected to the collection container 140 via the tube 160a. The culture medium and cell aggregates stored in the culture container 110 are classified by the suction tube nozzle 130-1, and the classified cell aggregates, together with the culture medium, are guided to the collection container 140 via the 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 area 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 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 a 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. A 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 a 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 container 110. The tube 160c and the pump 150 are in communication with each other.

[0092] <<<Circulation circuit>>> Except for the cell aggregates remaining at the suction tube nozzle 130-1, the cell aggregates move 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 in the suction tube nozzle 130-1>>> 2B is a cross-sectional view showing the outline of the movement of cell aggregates within the suction tube nozzle 130-1. In FIG. 2B, multiple white circular regions indicate cell aggregates.

[0097] <Top bus line UG and bottom bus line 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 generatrix lines along the longitudinal direction. Generatrix lines 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, an uppermost generatrix line UG located at the uppermost side of the suction tube nozzle 130-1 and a lowermost generatrix line BG located at the lowermost side of the suction tube nozzle 130-1 can be defined in the cross section of the suction tube nozzle 130-1. Note that the cross section of the suction tube nozzle 130-1 is a surface formed by cutting the suction tube nozzle 130-1 in a direction 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 position, 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 position, 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 case of tilted state> On the other hand, as shown in FIGS. 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 FIG. 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 generatrix UG and the lowermost generatrix 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 area 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 cell aggregates settle together with the culture medium. In particular, in the first flow region FF, cell aggregates tend to settle more easily in the region close to the bottom generating line BG. The cell aggregates settle faster in the region close to the bottom generating line BG and settle more slowly as they move away from the bottom generating line BG.

[0101] The culture medium flows in succession together with the cell aggregates through the suction opening 134-1. Therefore, the cell aggregates that settle near the suction opening 134-1 either flow out through the suction opening 134-1 or move to the second flow region SF and rise again. Whether they flow out through 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 area SF> The second flow region SF is a region where the cell aggregates also flow toward the discharge opening 136-1 as the culture medium rises 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 tend to rise more easily in regions close to the uppermost generating line UG. In other words, the cell aggregates rise faster in regions close to the uppermost generating line UG and rise more slowly as they move away from the uppermost generating line 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 culture 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 larger 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 larger 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 larger within the suction tube nozzle 130-1, they can be retained within the suction tube nozzle 130-1.

[0105] <<Discharged cell aggregates>> Due to the formation of the second flow region SF, cell aggregates smaller (lighter) than the reference diameter (size, mass) rise towards the discharge opening 136-1 and are discharged from the discharge opening 136-1 together with the medium.

[0106] <<Classification of cell aggregates>> Cell aggregates of a predetermined size or larger remain within the suction tube nozzle 130-1. On the other hand, cell aggregates smaller than the predetermined size are discharged from the discharge opening 136-1. In this way, by using the inclined suction tube nozzle 130-1, cell aggregates can be classified based on a predetermined diameter (size, mass) by forming a first flow region FF and a second flow region SF. Cell aggregates of a predetermined size or larger that remain within the suction tube nozzle 130-1 are discharged from the suction opening 134-1 of the suction tube nozzle 130-1. Cell aggregates smaller than the predetermined size are discharged from 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 cross section II shown in Figure 2A. In Figure 2B as well, multiple white circular regions 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 near the bottom generating line BG is occupied by cell aggregates, the area near 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 inclination of the suction tube nozzle 130-1 and the circulation flow rate>>> The relationship between the inclination of the suction tube nozzle 130-1 and the circulation flow rate is the relationship between the inclination angle θ of the suction tube nozzle 130-1 and the circulation flow rate obtained under the condition that cell agglomerates of a predetermined size or larger are retained in the suction tube nozzle 130-1 and discharged through the suction opening 134-1, and cell agglomerates of a size smaller than the predetermined size are discharged and recovered through 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 aspirate 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 aspirate 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 aspirate 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 collect 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] <<Cell aggregate recovery rate>> Fig. 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. Fig. 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 of a size 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 to the recovery vessel 140 through the suction tube nozzle 130-1. The recovery rate is the ratio of the cell aggregates recovered in the recovery vessel 140 that contain cell aggregates smaller than a predetermined (desired constant) size.

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

[0119] The relationship of the inclination angle θ of the suction tube nozzle 130-1 is 0 < θ1. As shown in FIG. 3, the relationship of the corresponding circulation flow rates is F0 < F1. That is, the circulation flow rate can be increased in the inclined state rather than in the vertical state.

[0120] Furthermore, as shown in FIG. 3, the relationship of the recovery amounts is V0 < V1. That is, the recovery amount has increased in the inclined state rather than in the vertical state.

[0121] Also, as shown in FIG. 3, the relationship of the recovery rates is C1 < C0. The recovery rate is higher in the vertical state than in the inclined state.

[0122] From the above, by setting the suction tube nozzle 130-1 in an inclined state rather than in the vertical state, it is possible to increase the recovery amount while maintaining the recovery rate to a certain extent. That is, by inclining the suction tube nozzle 130-1 and increasing the circulation flow rate to F1 by driving the pump 150, cell aggregates larger than a predetermined size are retained in the suction tube nozzle 130-1 and discharged from the suction opening 134-1, while cell aggregates smaller than the predetermined size are discharged from the discharge opening 136-1 of the suction tube nozzle 130-1, and the amount that can be recovered can be increased. In other words, by inclining the suction tube nozzle 130-1, the circulation flow rate can be increased, and the amount of cell aggregates that can be discharged from the suction tube nozzle 130-1 can be increased, so the time required for recovery can be shortened.

[0123] As described above, the relationship in FIG. 3 is determined on the condition that cell aggregates having a size greater than or equal to a predetermined (desired constant) size are retained in the suction tube nozzle 130-1 and discharged from the suction opening 134-1, and cell aggregates having a size less than the predetermined (desired constant) size are discharged from the discharge opening 136-1 of the suction tube nozzle 130-1 and collected. Therefore, if the size of the cell aggregates to be collected is changed, the inclination angle θ of the suction tube nozzle 130-1, the circulation flow rate, the collection amount, and the collection rate will be different. When collecting cell aggregates of various sizes, these conditions should be determined in advance through preliminary experiments or the like, and the inclination angle θ of the suction tube nozzle 130-1 and the circulation flow rate can be changed according to the size of the cell aggregates to be collected.

[0124] <<<Cell aggregate collection process by the collection system 100>>> The outline of the cell aggregate collection process by the collection system 100 will be described. Hereinafter, it is assumed that the cell aggregates are dispersed in the culture vessel 110 by the stirring device 120.

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

[0126] The cross-sectional area A2 of the collection vessel 140 is larger than the cross-sectional area A1 of the suction tube nozzle 130-1 (A1 < A2). Therefore, the flow velocity Q2 / A2 in the collection vessel 140 is smaller than the flow velocity Q1 / A1 in the suction tube nozzle 130-1. The cell aggregates introduced into the collection vessel 140 can remain in the collection vessel 140 by gravitational sedimentation. Due to the difference in flow velocity and the gravitational sedimentation of the cell aggregates, the culture medium can be discharged from the collection vessel 140 while the cell aggregates are retained in the collection vessel 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>>>> FIG. 4 is a schematic diagram showing the configuration of a collection system 200 according to the second embodiment. In FIG. 4, the same components as those in the first embodiment are denoted by the same reference numerals. In FIG. 4, 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. 4, the culture container 110 of the collection system 200 is fixedly arranged in a vertical state, unlike the collection system 100. The following mainly describes the differences from the collection system 100.

[0130] <Culture container 110> The structure 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 is an inclined portion 130-2a; a vertical portion 130-2b; a bending portion 130-2c; The suction tube nozzle 130-2 has an elongated shape as a whole, with an inclined portion 130-2a, a vertical portion 130-2b, and a bent portion 130-2c. The cross-sectional area of ​​the suction tube nozzle 130-2 is the area of ​​the inner diameter portion 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] <Slope part 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 disposed 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 portion 130-2b extends in the vertical direction. The vertical portion 130-2b has an elongated shape. The vertical portion 130-2b extends linearly along the longitudinal direction. The vertical portion 130-2b has a tubular shape. The vertical portion 130-2b has a discharge opening 136-1.

[0136] <Bending part 130-2c> The inclined portion 130-2a and the vertical portion 130-2b are connected via a 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, the inclined portion 130-2a extending in the inclined direction and the vertical portion 130-2b extending in the vertical direction may be connected by a curved member.

[0137] <Slanted part 130-2a, vertical part 130-2b, bent part 130-2c> The inclined portion 130-2a, the vertical portion 130-2b, and the bent portion 130-2c 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 surfaces of the inclined portion 130-2a, the vertical portion 130-2b, and the bent portion 130-2c. The inner circumferential portion constitutes the inner surfaces of the inclined portion 130-2a, the vertical portion 130-2b, and the bent portion 130-2c. The outer circumferential portion and the inner circumferential portion are positioned concentrically. A long surrounding region SR, surrounded by the inner circumferential portion 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 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 from 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 intake opening 134-1> Although only an example has been shown in which the inclined portion 130-2a is inclined linearly so that the suction opening 134-1 faces the side wall portion 112 of the culture vessel 110, 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 the third embodiment. In Figure 5, the same components as those 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 those in the first embodiment and are therefore omitted.

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

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

[0143] <Movable stand 370> The movable base 370 has: A culture vessel 110; a stirring device 120; a suction tube nozzle 130-1; It is equipped with:

[0144] The culture vessel 110, the agitator 120, and the aspirating tube nozzle 130-1 are provided so as 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 platform 370. By driving the actuator, the culture vessel 110, the agitator 120, and the aspirating tube nozzle 130-1 mounted on the movable platform 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 transitioned to the tilted state, the first flow region FF and the second flow region SF are formed, and the cell aggregates can be classified based on a predetermined diameter. 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 and tilted states> First, the processor of the control device (not shown) outputs a control signal to drive the actuator, displacing the movable base 370 and tilting the suction tube nozzle 130-1 (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 the pump 150, and sets the circulation flow rate to F1 (FIG. 3) (step S33). As a result, with the suction tube nozzle 130-1 inclined, more cell aggregates are stored in the 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, displaces the movable base 370, and sets the suction tube nozzle 130-1 to a vertical state (tilt angle 0 in FIG. 3) (step S37). Furthermore, the processor of the control device outputs a control signal to drive the pump 150, and sets 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 tilted state (tilt angle θ1), and sets the circulation flow rate to F1. The second predetermined time is the time it takes for the 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 aspirating 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 aspirating 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 aspirating 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, and therefore collection accuracy may decrease as the circulation flow rate increases. 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>>>> Fig. 6 is a schematic diagram showing the configuration of a collection system 400 according to the fourth embodiment. In Fig. 6, the same components as those in the first embodiment are denoted by the same reference numerals. In Fig. 6, 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.

[0162] 6, the recovery system 400 is fixedly arranged 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 is a suction tube nozzle 130-1; an expanded diameter portion 470; In other words, the suction tube nozzle 130-4 according to the fourth embodiment has a structure in which an expanded diameter portion 470 is provided at the discharge opening 136-1 of the suction tube nozzle 130-1 according to the first embodiment.

[0165] <Expanded diameter part 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] <Suction tube nozzle 130-1, expanded diameter portion 470, and surrounding region SR> The suction tube nozzle 130-1 and the 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 the expanded diameter portion 470. The inner circumferential portion constitutes the inner surface of the suction tube nozzle 130-1 and the expanded diameter portion 470. A long surrounding region SR, surrounded by the inner circumferential portion and extending along the longitudinal direction, functions as a hollow conduit. Culture medium and cell aggregates can flow through the surrounding region SR.

[0167] <Flow of 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 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 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 discharge opening 136-1 of suction tube nozzle 130-1 flow through enlarged diameter portion 470 and are discharged from discharge opening 472 of enlarged diameter portion 470.

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

[0170] <Inclined and vertical collection> Since 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), the suction tube nozzle 130-1 performs inclined collection, while the expanded diameter section 470 performs vertical collection. The inner diameter (cross-sectional area) of the expanded diameter section 470 is determined by the classification flow rate for vertical collection of cell aggregates of the 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 enlarged diameter section 470, it is possible to separate the role of enlarged diameter section 470 from that of aspirating tube nozzle 130-1, and cell aggregates can be collected efficiently.

[0172] <<<<Fifth embodiment>>>> Fig. 7 is a schematic diagram showing the configuration of a collection system 500 according to the fifth embodiment. In Fig. 7, the same components as those in the first embodiment are denoted by the same reference numerals. In Fig. 7, 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.

[0173] 7, the culture vessel 110 of the recovery system 500 is fixedly arranged in a vertical state, similar to the recovery system 200. Below, differences from the recovery system 100 will mainly be described.

[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 is a suction tube nozzle 130-2; 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] As in the fourth embodiment, the expanded diameter portion 470 has a discharge opening 472. The lower end of the expanded diameter portion 470 has an opening, which communicates with the discharge opening 130-2b.

[0177] <Suction tube nozzle 130-2, expanded diameter portion 470, and surrounding region SR> The suction tube nozzle 130-2 and the 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 the expanded diameter portion 470. The inner circumferential portion constitutes the inner surface of the suction tube nozzle 130-2 and the expanded diameter portion 470. A long surrounding region SR, surrounded by the inner circumferential portion and extending along the longitudinal direction, functions as a hollow conduit. Culture medium and cell aggregates can flow through the surrounding region SR.

[0178] <Flow of 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 expanded diameter portion 470 and are discharged from the discharge opening 472 of the expanded diameter portion 470.

[0179] <Inclined and vertical collection> The expanded diameter section 470 has an inner diameter larger than the inner diameter of the suction tube nozzle 130-2. That is, the expanded diameter section 470 has a cross-sectional area larger than the cross-sectional area of ​​the suction tube nozzle 130-2. By increasing the cross-sectional area of ​​the expanded diameter section 470, the flow rate in the expanded diameter section 470 can be reduced. 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 collection, while the expanded diameter section 470 performs vertical collection. The inner diameter (cross-sectional area) of the expanded diameter section 470 is determined by the classification flow rate for vertical collection of cell aggregates of the desired size. The suction tube nozzle 130-2 can store more cell aggregates, while the expanded diameter section 470 can perform 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 collection of cell aggregates.

[0181] <<<<Sixth embodiment>>>> FIG. 8 is a schematic diagram showing the configuration of a collection system 600 according to the 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 container 110> The structure 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] The suction tube nozzle 130-2, like the suction tube nozzle 130-1, An extension portion 132-1; an intake opening 134-1; a discharge opening 136-1; It has.

[0186] The extension portion 132-1 has a similar configuration to the aspirating 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 aspirating tube nozzle 130-1 extends in an inclined direction. In other words, the central axis AO of the aspirating 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 agitating blades 124a, and the central axis AO of the aspirating 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 agitating blades 124a extend vertically, and only the aspirating tube nozzle 130-1 extends in the inclined direction. The aspirating tube nozzle 130-1 is fixedly provided by a holding member or the like (not shown) so that it extends in a certain 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, by tilting the aspirating tube nozzle 130-2, the degree of freedom in the size and shape of the stirring blades 124a of the stirring bar 124 can be increased.

[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. On the other hand, 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 an inclined suction tube nozzle 130-2, the formation of the first flow region FF and the second flow region SF enables cell aggregates to be classified based on a predetermined diameter.

[0190] <<<<Seventh embodiment>>>> FIG. 9 is a schematic diagram showing the configuration of a collection system 700 according to the seventh embodiment. In FIG. 9, the same components as those in the first and second embodiments are denoted by the same reference numerals. In FIG. 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 FIG. 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] <Culture container 110-3> The structure and materials of the culture vessel 110-3 are basically the same as those of the collection system 100. In the collection systems 100 and 200, the culture 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 culture vessel 110. In contrast, the culture vessel 110-3 is formed integrally with the aspirating tube nozzle 130-3.

[0192] The suction tube nozzle 130-3 is An extension portion 132-3; an intake opening 134-3; a discharge opening 136-3; It has.

[0193] A through-hole is formed near the bottom 114 of the culture vessel 110-3. An intake opening 134-3 of the aspirating tube nozzle 130-3 is fixedly provided so as to communicate with the through-hole formed near the bottom 114 of the culture vessel 110-3. The aspirating 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 become more distant from the culture vessel 110-3 as it extends upward. The central axis AO of the aspirating tube nozzle 130-3 extends in the inclined direction.

[0194] By forming the aspirate tube nozzle 130-3 integrally with the culture vessel 110-3, a holding member for holding the aspirate tube nozzle 130-3 is not required. Also, since the tilt angle θ of the aspirate tube nozzle 130-3 can be kept constant, adjustment of the tilt angle θ is also not required. Furthermore, because the aspirate tube nozzle 130-3 is disposed outside the culture vessel 110-3, the aspirate tube nozzle 130-3 does not interfere with the stirring blades 124a of the stirring bar 124. This configuration allows the medium and cell aggregates to be sufficiently stirred, and increases the degree of 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 in 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. On the other hand, 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 an inclined suction tube nozzle 130-3, the formation of the first flow region FF and the second flow region SF enables cell aggregates to be classified based on a predetermined diameter.

[0197] <Other Configurations of Culture Vessel 110-3> 9 shows a configuration in which the aspirating tube nozzle 130-3 is integrally formed 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 substantially 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 configuring the aspirate tube nozzle 130-3 separately from the culture vessel 110-3, it is possible to use aspirate tube nozzle 130-3 and culture vessel 110-3 made of different materials, or to use aspirate tube nozzle 130-3 having a desired length or shape.

[0199] Furthermore, the positions of the through-holes and protrusions for attaching the aspirating tube nozzle 130-3 and the angles thereof relative to the vertical direction 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 embodiment>>>> 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 here are also included. [Industrial Applicability]

[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. [Explanation of symbols]

[0202] 100, 200, 300, 400, 500 Collection System 110 Culture vessel 130-1, 130-2, 130-4, 130-5 Suction tube nozzle 132-1 Extension

Claims

1. a culture section in which a fluid containing cell aggregates is accommodated; A classification system comprising a classification unit that classifies 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 unit including either an extension unit in which the fluid flows along the 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 unit that is fixedly inclined.

2. The classifying unit includes: an intake opening for intake of the fluid from the culture unit; a discharge opening spaced apart from the suction opening along the extension direction of the classification unit and through which the first cell aggregate is discharged; The classification system of claim 1 , comprising:

3. The classification system according to 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 unit.

4. In the inclined extension portion of the classification portion, a first flow region in which the second cell aggregates flow toward the intake opening by reciprocating movement; a second flow region in which the second cell aggregates flow through the first flow region, causing the first cell aggregates to flow toward the discharge opening; The classification system of claim 3 , wherein

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 at least the straight pipe section has an angle that can be changed with respect to the vertical direction between a first angle and a second angle that is different from the first angle.

7. the classification section has a bent portion through which the fluid can flow, and includes a first classification section and a second classification section that are connected via the bent portion and extend in different directions from each other, The classification system according to claim 1 , wherein one of the first classifying section and the second classifying section is an inclined extension of the classifying section.

8. 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 portion has the intake opening; The classification system of claim 2 , wherein the large diameter portion includes the discharge opening.

Images