Cell recovery device and cell recovery method

Abstract

The present invention is a cell recovery device characterized by comprising: a stage provided with a plurality of containers each having a cap that engages with a container body, a capper for performing an operation for engaging or separating each cap and the corresponding container body, a pipette member for performing a liquid suction operation and a liquid discharge operation in a container body, a robot for selectively gripping and operating the containers, and a cap loading part for temporary loading of respective caps removed from the container bodies; a control unit capable of controlling actuation of the robot, the operations for engagement and separation by the capper, and the operations for liquid suction and liquid discharge by the pipette member; and a safety cabinet for providing an aseptic working environment, the stage being positioned inside the safety cabinet, whereby the cap loading part, the capper, the pipette member, and the robot are all arranged in said working environment.

Description

[Technical Field]

[0001] The present invention relates to cell culture, and more particularly to a cell recovery device and a cell recovery method using the cell recovery device. [Background technology]

[0002] In cell culture, cells cultured in various culture vessels must be recovered for subculture. In conventional techniques, such cell recovery from culture vessels is almost always performed manually by an operator. However, since a certain amount of cells is required for cell-based products, cell culture and subculture must be performed multiple times. Because a long time required for cell subculture can damage the cells, the subculture procedure must be completed in a short time. Furthermore, to prepare cells of a consistent quality, the same procedure must be performed in each step. In addition, strict requirements for sterility during the procedure mean that manual procedures require a high level of skill, are complicated, require a lot of man-hours, and are prone to errors. Summary of the Invention [Problem to be solved by the invention]

[0003] The present invention has been made in view of the above-mentioned problems of the conventional technology, and aims to provide a cell recovery device and a cell recovery method that can prevent errors from occurring in cell recovery operations during cell culture by automating at least a portion of the operations that are currently performed manually by humans using a robot.

[0004] In order to achieve the above object, the following inventions have been adopted. [Means for solving the problem]

[0005] The present invention provides a method for manufacturing a container, comprising: a stage provided with a capper that performs an operation of engaging or separating the cap with the corresponding container body, a pipette member that performs an operation of suctioning or discharging liquid from the container body, a robot that selectively grasps and operates the container, and a cap placement unit that temporarily places each of the caps removed from each of the container bodies; a control unit capable of controlling the operation of the robot, the engaging operation and the separating operation of the capper, and the liquid suction operation and the liquid discharge operation of the pipette member; A cell recovery device is provided that includes a safety cabinet that provides a sterile working environment, and the stage is disposed within the safety cabinet, so that the cap mounting portion, the capper, the pipette member, and the robot are all located within the working environment.

[0006] Preferably, the cell collection device further includes a cap state detector capable of detecting whether the container is covered with the cap.

[0007] More preferably, the cell collection device includes a movable container unit that holds the container body so as to tap the container body and / or tilt the container body relative to the stage. Note that "and / or" means at least one of the two, and includes either one or both.

[0008] More preferably, the tapping is a reciprocating vibration operation that reciprocates once or twice per second for a period of 10 seconds or more.

[0009] More preferably, the container further includes a container body holder for fixedly holding the container body, and the container body holder has a gripping portion that is gripped by the robot.

[0010] More preferably, the portion of the capper that grips the cap has two or more inner diameters, so that the capper can grip and open / close caps having different diameter ranges.

[0011] More preferably, the cap placing section has a cap receiving member that can be grasped by the robot and that receives the cap separated from the container body.

[0012] The present invention provides a cell recovery method using the cell recovery device according to any one of the above aspects, In the work process controlled by the control unit, a culture medium discharging step of discharging the culture medium in a culture vessel, which is one of the vessels; a washing step of injecting a cell washing solution into the culture vessel and then discharging the cell washing solution; a detachment step of injecting a cell detachment solution into the culture vessel to detach cells in the culture vessel; a washing step of washing the cells in the culture vessel; a recovery step of discharging the cells in the culture vessel into a cell recovery bottle, which is one type of the vessel; The present invention further provides a cell collection method that sequentially includes a suspension step in which the pipette member repeatedly performs the liquid suction operation and the liquid discharge operation on the cell collection bottle, thereby suspending the cells in the cell collection bottle.

[0013] Preferably, the method further includes an additional recovery step of injecting a cell washing solution into the culture vessel after the recovery step and before the suspension step, and then injecting the remaining cells into the cell recovery bottle using the cell washing solution.

[0014] More preferably, the cap state detector performs detection after every engagement or separation operation involving the cap.

[0015] More preferably, in any of the washing step, the peeling step, the recovery step, and the additional recovery step, the shaking operation of the culture vessel is performed via the robot.

[0016] More preferably, at least one of all the operations relating to the robot includes an operation of moving the robot to a reset position.

[0017] The present invention also provides a method for manufacturing a container having a plurality of containers each having a cap that engages with a container body; a capper that performs an engagement or separation operation between the cap and the container body; a pipette member for sucking and discharging liquid into and from the container body; a robot that selectively grasps and operates a container from among the plurality of containers; a cap placement portion for placing each of the caps separated from each of the container bodies; The present invention provides a cell recovery device that includes a control unit that can control the operation of the robot, the engagement and separation operations of the capper, and the liquid intake and discharge operations of the pipette member.

[0018] Preferably, a stage on which the capper, the pipette member, the robot, and the cap placement unit are provided; and a safety cabinet that forms a sterile working environment therein and has the stage disposed therein.

[0019] The present invention provides a cell collection device and a cell collection method using the cell collection device, as described below. The cell collection device includes a stage and a control unit. The stage is provided with a capper that operates the caps of different containers to engage or separate the caps from their corresponding container bodies, a pipette member that performs aspirating and dispensing operations on the container bodies, a robot that selectively grasps and operates the containers, and a cap placement unit that temporarily places each cap removed from each container body. With the above-described structure, the control unit can accurately control the capper's engagement and separation operations and the pipette member's aspirating and dispensing operations with precise amounts according to a predetermined program, and can also accurately control the robot's movements. This allows at least some of the cell collection operations for cell culture to be automated using a robot instead of manual operations, thereby preventing errors in the cell collection operation. [Effects of the Invention]

[0020] According to the present invention, it is possible to prevent mistakes from occurring during the cell recovery process. [Brief explanation of the drawings]

[0021] [Figure 1] Fig. 1 is a schematic front view showing a cell collection device according to one embodiment of the present invention, in which a safety cabinet is omitted. [Figure 2] 2 is a schematic plan view showing a cell collection device according to one embodiment of the present invention, in which the safety cabinet is omitted. [Figure 3] FIG. 3 shows the first bottle held in the container body holder. [Figure 4] FIG. 4 shows the first bottle held in the container body holder. [Figure 5] FIG. 5 shows a state in which the container body holder holding the first bottle is held by the robot holder. [Figure 6] FIG. 6 shows the state in which the second bottle is held by the container body holder. [Figure 7] FIG. 7 shows the state in which the second bottle is held by the container body holder. [Figure 8] FIG. 8 shows the state in which the container body holder holding the second bottle is held by the robot holder. [Figure 9] FIG. 9 shows the state in which the third bottle is held by the container body holder. [Figure 10] FIG. 10 shows the state in which the container body holder holding the third bottle is held by the robot holder. [Figure 11] FIG. 11 shows the state in which the fourth bottle is held by the container body holder. [Figure 12] FIG. 12 shows the state in which the container holder holding the fourth bottle is held by the robot holder. [Figure 13] Fig. 13 is a schematic front view showing the safety cabinet of a cell collection device according to one embodiment of the present invention, and is a schematic view showing the exterior of the safety cabinet. [Figure 14] 14 is a schematic side view showing the safety cabinet of a cell collection device according to one embodiment of the present invention. The schematic side view of FIG. 14 shows the interior of the safety cabinet. DETAILED DESCRIPTION OF THE INVENTION

[0022] Hereinafter, exemplary embodiments of the present invention will be described with reference to the drawings. It should be understood that these specific descriptions are intended merely to teach those skilled in the art how to implement the present invention, and do not enumerate all possible embodiments of the present invention, nor are they intended to limit the scope of the present invention. In this specification, the term "vertical direction" refers to the direction of gravity (the up-down direction in FIG. 1 ), and the term "horizontal direction" refers to the horizontal direction in a plane perpendicular to the vertical direction (the left-right direction in FIG. 1 ). Furthermore, "tilting" primarily refers to the process of tilting the central axis of a container at a certain angle relative to the vertical direction via a movable container (described below). "Tapping" primarily refers to the reciprocating motion of the container via a movable container (described below) at a predetermined posture and frequency within a predetermined range for a predetermined time, thereby achieving reciprocating vibration of the container. "Shaking" primarily refers to the combined motion process of reciprocatingly shaking and rocking the container within a certain range via a robot (described below). During this motion process, the centerline of the container may extend along the vertical direction, the horizontal direction, or may not extend along a specific direction.

[0023] The structure of a cell recovery device according to one embodiment of the present invention will be described below with reference to the drawings. (Structure of a cell collection device according to one embodiment of the present invention)

[0024] As shown in Figures 1 to 14, a cell recovery device according to one embodiment of the present invention comprises a safety cabinet 6, a control unit CU provided outside the safety cabinet 6, and a plurality of containers C, a stage 1, a mounting member 2, a capper 3, a pipette member 4 and a robot 5 provided inside the safety cabinet 6.

[0025] In this embodiment, the containers C contain fluids such as cell recovery culture media and cell solutions, and can be grasped by the robot 5 and moved to various positions. As shown in Figures 3, 4, 6, 7, 9, and 11, each container C has a container body CB that forms a storage space, a cap CL for closing the opening of the container body CB, and a container body holder CH that securely holds the container body CB. Specifically, in this embodiment, the containers C are divided into a first bottle C1, a second bottle C2, a third bottle C3, and a fourth bottle C4.

[0026] The first bottle C1 has a square cross section of the container body CB, a volume of 125 ml, and is a bottle containing a cell washing solution or a cell detachment solution. The cell washing solution may be, for example, PBS (phosphate buffered saline), and the cell detachment solution may be, for example, trypsin. Before the cell recovery device of the present invention begins operation, the first bottle C1 is accommodated in a corresponding one of the placement positions formed by the movable container unit 212. The movable container unit 212 is capable of rotating around an axis extending along the vertical direction V and of linear displacement along the vertical direction V. The container body holder CH of the first bottle C1 is provided with a gripping portion that is gripped by the container holder 542 of the robot 5.

[0027] The second bottle C2 has a circular container body CB with a volume of 500 ml. The lower half of the container body CB of the second bottle C2 tapers toward the bottom. The second bottle C2 may be a container, such as a centrifuge tube for cell collection. The second bottle C2 may further have other structures that are advantageous for concentration, flow, distribution, and collection of the cell solution. Before the cell collection device according to the present invention starts operating, the second bottle C2 may be accommodated in the mounting position of the movable container 212 or the fixed container 211. For example, before the cell collection device starts operating, the second bottle C2 is accommodated at position Pa (see FIG. 2). The container body holder CH of the second bottle C2 is provided with a gripping portion that is gripped by the container holder 542 of the robot 5.

[0028] The third bottle C3 is a bottle whose container body CB has a rectangular cross section. Before the cell recovery device according to the present invention starts operating, the third bottle C3 is accommodated in a corresponding mounting position of the fixed storage unit 211, and is thereby fixed to the stage 1. For example, before the cell recovery device starts operating, the third bottle C3 is accommodated in position Pb (see FIG. 2). The third bottle C3 is used as a culture vessel to accommodate culture medium and grown cells. The cells are eventually concentrated in the second bottle C2. The container body holder CH of the third bottle C3 is provided with a gripping portion that is gripped by the container holder 542 of the robot 5.

[0029] The fourth bottle C4 has a square cross section of the container body CB and a volume of 2000 ml. Before the cell recovery device according to the present invention starts operating, the fourth bottle C4 is accommodated in a corresponding mounting position of the fixed storage unit 211, and is thereby fixed to the stage 1. For example, before the cell recovery device starts operating, the fourth bottle C4 is accommodated in position Pc (see FIG. 2). The fourth bottle C4 is a bottle that accommodates the discharged culture medium, and all of the culture medium discharged from the third bottle C3 is collected in the fourth bottle C4.

[0030] The container body holders CH for the four bottles C1, C2, C3, and C4 described above may be dedicated resin brackets or artificially formed silicone rubber brackets. This configuration prevents the bottles from falling even when they are inverted. Furthermore, because the bottles C1, C2, C3, and C4 are made of plastic, when the robot 5 directly grips the container body CB, bending can make it difficult to operate the caps and the bottles can easily fall. However, the container body holders CH can prevent these problems.

[0031] In this embodiment, as shown in FIGS. 1 and 2, the stage 1 is installed on a work table 62 (see FIG. 14) inside a safety cabinet 6. The stage 1 has a work surface 1a extending along the horizontal direction H. A mounting member 2, a capper 3, a pipette member 4, and a robot 5 are installed on the work surface 1a.

[0032] Specifically, in this embodiment, the mounting member 2 is provided for mounting the container body CB and cap CL of the container C. The mounting member 2 has a container body accommodating section 21 and a cap mounting section 22. The container body accommodating section 21 and the cap mounting section 22 form mounting positions corresponding to the container body CB and cap CL of the container C, respectively, a temporary storage position for temporarily storing the container C, and a tilting / swinging position for tilting the container body CB of the container C (normally, when the cap CL is not engaged) or performing a reciprocating motion (i.e., tapping) (normally, when the cap CL is engaged).

[0033] Furthermore, the container body accommodating section 21 accommodates the container body CB of the corresponding container C and has an accommodating space that matches the cross-sectional shape of the container body CB. The multiple container body accommodating sections 21 are distributed on the stage 1 so as to surround the robot 5, thereby facilitating the robot 5 to operate on the container C inside the container body accommodating section 21. The container body accommodating section 21 has a fixed accommodating section 211 that cannot move relatively and a movable accommodating section 212 that can move relatively. Note that the "corresponding container C" refers to the container C accommodated in the container body accommodating section 21, such as a bottle C1.

[0034] The fixed storage part 211 holds its corresponding container body CB so that it is fixed relative to the work surface 1a. In this embodiment, the fixed storage part 211 holds its corresponding container body CB so that its central axis is perpendicular to the work surface 1a. In other words, the central axis of the container body CB extends along the vertical direction V. In this way, the opening of the container body CB stored in the fixed storage part 211 always faces upward in the vertical direction V. The fixed storage part 211 forms the above-mentioned placement position and temporary storage position. Note that the "corresponding container body CB" refers to the container body CB of the container C held by the fixed storage part 211, for example, the container body CB of the bottle C4.

[0035] The movable container 212 holds the corresponding container body CB so that it can rotate relative to the work surface 1a. In this embodiment, the movable container 212 for accommodating the first bottle C1 is rotatable about an axis extending along the vertical direction V and linearly displaceable along the vertical direction V, allowing the first bottle C1 to be positioned at different positions in a predetermined vertical direction V and different positions in the horizontal direction H. If necessary, containers C other than the first bottle C1 can be placed in the movable container 212, allowing each container C to be moved to a position convenient for manipulation by the robot 5. Furthermore, the movable container 212 also forms the tilting / swinging position, allowing the container C moved to the tilting / swinging position by the robot 5 to perform desired tilting and / or reciprocating motion in the movable container 212. The movable container 212 for forming the tilting / swinging position accommodates a specific container C only when necessary, thereby tilting the central axis of the container body CB of the container C relative to the horizontal work surface 1a and / or performing a reciprocating motion of the container C within a predetermined range, thereby promoting uniform mixing and distribution of the solution in the container C. Note that the "corresponding container body CB" refers to the container body CB of the container C held by the movable container 212, for example, the container body CB of the bottle C1.

[0036] Furthermore, the cap placing unit 22 forms a placing position for placing the cap CL by placing the cap CL removed from the container C. In this embodiment, the cap placing unit 22 has a cap receiving member 221 that can be clamped by the robot. The cap receiving member 221 is formed with a plurality of recesses 222 that match the shape of each cap CL, so that the recesses 222 can accommodate the corresponding cap CL, and each recess 222 corresponds to the placing position of one cap CL. The cap receiving member 221 can be moved from the stage 1 to the capper 3 by the robot 5, and the cap receiving member 221 places the cap CL in the recess 222 by aligning the corresponding recess 222 with the cap CL removed by the capper 3. In addition, the cap receiving member 221 can be moved from the stage 1 to the capper 3 by the robot 5, and the cap receiving member 221 aligns the cap CL placed on the cap receiving member 221 with the capper 3, thereby engaging the capper 3 with the cap CL and then tightening the cap CL to its corresponding container body CB.

[0037] In this embodiment, the capper 3 is provided to operate the caps CL of different containers C to engage and separate the caps CL and their corresponding container bodies CB. The phrase "different containers C" refers to the ability to operate multiple containers C separately, meaning that the capper 3 can operate one container C and then another different container C. The capper 3 has an engagement claw 31 that can engage with the cap CL of a container C and rotate in two directions around the vertical direction V. The engagement claw 31 has three claw portions 311 evenly distributed around the circumferential direction of the cap CL, allowing it to fasten or remove the cap CL of the container C. After engaging with the cap CL attached to the container body CB of the container C from the outside, the engagement claw 31 can be rotated in one direction to remove the cap CL from the container body CB and separate the cap CL from the container body CB. Meanwhile, the engagement claw 31 can close the opening of the container body CB by engaging the cap CL with the container body CB.

[0038] The capper 3 also has a cap state detector (not shown) that detects whether the opening of the container C is covered with the cap CL. The cap state detector may be an optical sensor. In the cell recovery method described below, after performing the engagement or separation operation for all caps CL, the cap state detector detects whether the caps CL have been securely engaged with or separated from the container body CB. If it is detected that the desired state has not been achieved, the control unit CU controls the buzzer to issue an alarm and stops the operation.

[0039] The capper 3 may also be provided with a clamping module that clamps the container body CB of the container C. The clamping module has two clamping sections that approach or separate from each other, thereby fixing and holding the container body CB of the container C with the two clamping sections and relatively fixing the container body CB. In this way, when the capper 3 operates the cap CL of the container C to engage or separate the cap CL from the container body CB, the container body CB does not rotate freely due to the operation of the engagement claw 31 of the capper 3. Furthermore, the inner peripheral wall of the engagement claw 31 may be formed with engagement portions having two or more inner diameters. This allows the engagement claw 31 to engage with caps CL of different diameter ranges, thereby enabling it to clamp caps CL of different diameter ranges. For example, if the inner diameters of the engagement portions on the inner peripheral wall of the engagement claw 31 are 5 cm and 7 cm, it can engage with caps CL with diameters of 5 to 6 cm and also with caps CL with diameters of 7 to 8 cm. Of course, any design may be used as long as the inner diameter dimensions are different, and the specific dimensional range can be adjusted as needed.

[0040] In this embodiment, the pipette member 4 includes an assembled pipette (50 ml), a silicone hose, and a glass syringe. The pipette enters the container body CB and performs direct suction (liquid suction) and discharge (liquid discharge) operations on the solution in the container body CB. The silicone hose is located between the pipette and the glass syringe and has a certain degree of flexibility. It serves to airtightly connect the pipette and the glass syringe, thereby preventing the pipette from being hindered from moving in the vertical direction V due to the connection being unable to deform. The pipette and silicone hose are both disposable. The glass syringe functions as a piston, providing power for the suction and discharge operations. In this way, the pipette member 4 suspends the cells in the cell solution collected in the second bottle C2 (dispersing, homogenizing, and suspending the cells). The pipette member 4 performs the above-mentioned suspension process by suctioning (suctioning) the solution from the container body CB of the container C or discharging (discharging) the solution into the container body CB of the container C in different amounts.

[0041] In this embodiment, the robot 5 has a mechanical structure with six degrees of freedom and is configured to move the container body CB of the container C and the cap receiving member 221. The robot 5 includes multiple arms 51, 52, 53, and 54 connected in series. Among these, a container holder 542 (holding portion) at the end of the fourth arm 54, which is the terminal arm, can hold the container C in at least two positions (corresponding to a first holding mode and a second holding mode). When the container holder 542 holds the container C in the first holding mode, the container holder 542 of the fourth arm 54 and the central axis of the container body CB of the container C form a first angle. When the container holder 542 holds the container C in the second holding mode, the container holder 542 of the fourth arm 54 and the central axis of the container body CB of the container C form a second angle. The difference between the first angle and the second angle may be any value greater than 0° and less than 180°, and causes the container C to perform a predetermined motion (linear displacement and swinging). The fourth arm 54 has a base 541 extending along a first direction and a holder 542 extending along a second direction perpendicular to the first direction. The base 541 and the container holder 542 are fixedly connected. The base 541 is connected to the third arm 53 described below, and the holder 542 is provided to hold the container C. The holder 542 can hold the container C so that the central axis of the container C coincides with the first direction or the second direction, thereby holding the container C via the container body holder CH in the first holding mode or the second holding mode. In this case, the difference between the first angle and the second angle is 90°. When the central axis of the container C coincides with the first direction (for example, the configurations shown in FIGS. 5, 8, 10, and 12), the robot 5 mainly moves the container C horizontally and linearly with the opening facing upward, i.e., moves the container C between different positions. When the central axis of the container C coincides with the first direction or the second direction, the robot 5 can tilt the opening of the container C at any angle with respect to the vertical direction V to tilt the solution in the container body CB, or can hold and shake the container C to uniformly mix the solution in the container C.Tilting the solution in the container C in two relative positions has an effect similar to that of holding the container C in the forward and backward directions with a human arm, allowing the solution in the container C to be tilted more completely and preventing excessive residual solution. Rocking the container C in two relative positions allows the solution in the container C to be mixed more uniformly. Furthermore, in this embodiment, the holder 542 has a first holding portion and a second holding portion located in different positions. The first holding portion and the second holding portion can each hold the same portion of the container C. The first holding portion is in the first holding mode when holding a held object, and the second holding portion is in the second holding mode when holding a held object. Furthermore, the holder 542 has a connection pin 542P connected to the container C, and holds the container C by clamping the connection pin 542P. The connection pin 542P can engage with a holding hole pre-formed in the container C, thereby fixing the holder 542 relative to the container C. In this embodiment, two connection pins 542P are provided for each group of connection pins 542P, and one group of connection pins 542P and another group of connection pins 542P are arranged to protrude in opposing directions.

[0042] Furthermore, excluding the fourth arm 54, the multiple arms 51, 52, and 53 of the robot 5 include a first arm 51 having one end attached to the stage 1 of the cell recovery device, a second arm 52 connected to the first arm 51, and a third arm 53 connected to the second arm 52. The first arm 51 extends along a vertical direction V and is rotatable around the vertical direction V. The second arm 52 extends linearly and has one end connected to the other end of the first arm 51 via a first joint 51J, which provides the second arm 52 with at least one degree of rotational freedom relative to the first arm 51. The third arm 53 extends linearly and has one end connected to the other end of the second arm 51 via a second joint 52J, which provides the third arm 53 with at least one degree of rotational freedom relative to the second arm 52. The fourth arm 54 is connected to the third arm 53 via a third joint 53J, and the third joint 53J provides the fourth arm 54 with at least one degree of rotational freedom relative to the third arm 53. Specifically, the first joint 51J is provided with a first axis extending along the horizontal direction H, and the second arm 52 is rotatable around the first axis. The second joint 52J is provided with a second axis extending along the horizontal direction H, and the third arm 53 is rotatable around the second axis. The third joint 53J is provided with a third axis extending along the horizontal direction H, and the fourth arm 54 is rotatable around the third axis. Furthermore, the third arm 53 and the fourth arm 54 are both rotatable around their respective central axes. In this way, the robot 5 can achieve six degrees of freedom of movement.

[0043] According to the above-described invention, the robot 5 has a mechanical structure with six degrees of freedom, which enables it to hold other members in desired positions and complete tasks such as linear displacement, rotation, tilting, and swinging of the container C. Specifically, the robot 5 can smoothly complete each desired task by moving each container C to the capper 3, aligning each recess 222 of the cap receiving member 221 with the removed cap CL, and moving each container C to a placement position, a temporary storage position, a tilted position, etc.

[0044] In this embodiment, the control unit CU includes a CPU, memory, etc., and is connected to the robot 5 via a wired or wireless connection so as to be able to send and receive control signals. The control unit CU controls the robot 5 to perform various operations on the container C, controls the engagement and separation operations of the capper 3, and can control the selective suction and discharge operations of the pipette member 4, in accordance with a predetermined program stored in the memory. Although not clearly explained in this embodiment, the control unit CU may have multiple control means for controlling the operation of each member, thereby completing each of the above operations in accordance with the predetermined program.

[0045] 13 and 14, a safety cabinet 6 is provided to provide a sterile working environment. The stage 12 may be installed on a detachable work table 62 inside the safety cabinet 6, or may be the same member as the work table 62. The safety cabinet 6 has a housing 61, an exhaust module 63, a measurement module 64, a sterilization module 65, an illumination module 66, and a movement adjustment module 67.

[0046] Specifically, in this embodiment, the housing 61 has a cubic shape and forms a sterile working environment therein. The work table 62 is attached within the housing 61 and is arranged horizontally.

[0047] Furthermore, in this embodiment, an exhaust module 63 is provided to communicate between the inside and outside of the housing 61. The exhaust module 63 has an air inlet 631 and an exhaust outlet 635 provided in the housing 61, an air intake filter 632 (two in this embodiment) provided in the housing 61 and positioned at the air inlet 631, and an exhaust filter 634 (one in this embodiment) provided in the housing 61 and positioned at the exhaust outlet 635, thereby filtering out dust, particles, etc. during gas exchange between the inside and outside of the housing 61. The exhaust module 63 further has a blower 633 provided inside the exhaust outlet 635. The blower 633 is provided to promote the exhaust of air inside the housing 61 to the outside. The exhaust module 63 also has an exhaust filter differential pressure gauge 636 that monitors the pressure difference of the exhaust filter 634.

[0048] Furthermore, in this embodiment, a measurement module 64 is provided to measure predetermined parameters within the housing 61. Specifically, the measurement module 64 has a particle counter 641 that measures the concentration of dust and particles within the housing 61, and a PAO (polyalphaolefin) inlet side measurement valve 642 that measures the input of PAO, thereby making it possible to provide the user with desired parameter information.

[0049] Furthermore, in this embodiment, sterilization module 65 is provided in housing 61 and is provided to sterilize the inside of housing 61. Specifically, sterilization module 65 has two germicidal lamps (15 W) provided on the side walls of housing 61, thereby making it possible to effectively sterilize the internal space of housing 61.

[0050] Furthermore, in this embodiment, an illumination module 66 is provided to illuminate the interior of the housing 61. Specifically, the illumination module 66 includes three LEDs provided at the top of the interior space of the housing 61.

[0051] Furthermore, in this embodiment, the movement adjustment module 67 is disposed under the housing 61 and is provided for moving the housing 61 and adjusting the angle between the housing 61 and the ground. Specifically, the movement adjustment module 67 has a plurality of rollers 671, a plurality of adjusters 672, and a plurality of support legs 673. The rollers 671 are advantageous for the housing 61 to move along the ground when moving. The adjusters 672 are provided for supporting the housing 61 while adjusting the angle between the housing 61 and the ground. The support legs 673 perform a supporting function for the housing 61 after being placed down.

[0052] Furthermore, in this embodiment, the operation display unit 68 is arranged on the outer wall of the housing 61 and is provided to display parameters within the housing 61 and to input operation commands, etc., that control the operation of each component of the safety cabinet 6.

[0053] The above is a description of the structure of a cell recovery device according to one embodiment of the present invention. Below, we will explain the automated operation steps (cell recovery method) of the cell recovery device using examples. The following operation steps can all be automatically completed by the cell recovery device according to the present invention in accordance with a predetermined program. (Example of operation steps (working process) of a cell recovery device according to one embodiment of the present invention)

[0054] The operation steps of the cell recovery device according to one embodiment of the present invention are as follows: a culture medium discharging step of discharging most of the culture medium in the third bottle C3, which is a culture vessel, into the fourth bottle C4 via the robot 5; a washing step in which the cell washing solution contained in the first bottle C1 is poured into the third bottle C3 via the robot 5 to completely wash away the remaining culture medium with the cell washing solution, and then the cell washing solution is discharged into the fourth bottle C4, in which the third bottle C3 needs to be tilted to one side via the movable container 212 to facilitate the tilting operation when the cell washing solution is poured into the third bottle C3; a detachment step in which the cell detachment solution contained in the first bottle C1 is poured into the third bottle C3 via the robot 5 to detach the cells in the third bottle C3, in which the third bottle C3 needs to be tilted to one side via the movable container 212 to facilitate the tilting operation when the cell detachment solution is poured into the third bottle C3; a washing step of washing the cells with the cell washing solution by injecting the cell washing solution into a third bottle C3 via the robot 5; a recovery step of discharging the cell detachment solution and cells in the third bottle C3 into the second bottle C2, which is a cell recovery bottle, via the robot 5; and a suspension step of suspending the cells in the second bottle C2 via the pipette member 4.

[0055] Furthermore, the cell recovery method further includes a supplementary recovery step after the recovery step and before the suspension step, in which a cell washing solution is injected into the third bottle C3, and then the remaining cells are injected into the second bottle C2 using the cell washing solution, thereby enabling all of the cells remaining in the third bottle C3 to be recovered as completely as possible into the second bottle C2 (additional recovery step).

[0056] Furthermore, in all of the above-described washing step, peeling step, recovery step, and replenishment / recovery step, the third bottle C3 is rocked via the robot 5. The rocking is performed with the thickness direction of the third bottle C3 aligned substantially vertically, and the culture vessel is inverted upside down at least once during the rocking operation. This upside-down inversion is performed so as not to affect the substantially vertical state of the thickness direction. In the above-described recovery step and replenishment / recovery step, the third bottle C3 can be placed on the movable container 212 during the rocking operation by the robot 5, and the third bottle C3 can be reciprocally displaced via the movable container 212. The reciprocating displacement is a reciprocating vibration operation that continues for, for example, 10 seconds or more, with one to two reciprocating movements per second. In this vibration operation, the reciprocating movement is suddenly stopped after accelerating, simulating a tapping-like effect, allowing the cells in the third bottle C3 to fully enter the solution and thoroughly mixing the solution in the third bottle C3, thereby achieving complete recovery.

[0057] Furthermore, the tilting operation of the robot 5 is performed in each of the above-described steps. In the tilting operation, the liquid in the container C is drained while the container C is held in the first holding mode and the second holding mode, respectively. In this way, the liquid in the container C can be drained as completely as possible. Furthermore, in each of the above-described steps, it is preferable to provide an operation of moving the robot 5 to a reset position between all of the robot 5's operations. This is advantageous for setting the robot 5 to the reset position for the next operation and for troubleshooting if the robot 5 malfunctions. After the malfunction is resolved, the next operation can be performed from the predetermined reset position for that operation. Here, although an operation of moving the robot 5 to a reset position is provided between all of the robot 5's operations, an operation of moving the robot 5 to a reset position may be provided only between some of the robot 5's operations. Furthermore, during most of the suspension process, the pipette member 4 enters below the surface of the liquid in the second bottle C2 to perform the aspirating and dispensing operations, thereby performing the suspension process. At the end of the suspension process, the pipette member 4 is moved away from the liquid surface in the second bottle C2, but by continuing to perform the suction and discharge operations in the air, the cells remaining in the pipette member 4 can be completely injected into the second bottle C2.

[0058] Therefore, the present invention provides a novel cell collection device, which is not limited to the examples described in the above specific embodiments, and further includes the following supplementary explanation.

[0059] (i) In the specific embodiment described above, the working surface 1a of the stage 1 is horizontal, but this is not limited to the present invention. If necessary, the working surface 1a may be set to form a predetermined angle with respect to the horizontal plane.

[0060] (ii) In the specific embodiment described above, the structure of the safety cabinet 6 of the cell collection device according to the present invention has been described, but the present invention is not limited to this. The safety cabinet 6 may have a desired power socket and power cord. Furthermore, the housing 61 may have a detachable structure for maintenance and a transparent structure for easy observation of the internal space.

[0061] (iii) In the specific embodiment described above, the robot 5 of the cell recovery device according to the present invention has six degrees of freedom, but the present invention is not limited to this. The robot 5 may be a redundant robot with seven degrees of freedom.

[0062] (iv) In the specific embodiment described above, an example was described in which the cap CL is gripped via the engaging claws 31, but the member for gripping the cap CL of the capper 3 is not limited to a claw and may be another type of gripping portion. The other type of gripping portion may be set to have multiple inner diameters that can be applied to multiple gripping size ranges.

[0063] (V) In the specific embodiment described above, an example is described in which the movable housing part 212 can realize a reciprocating vibration motion that imitates tapping and can also realize tilting, but the movable housing part 212 may also realize different movements using different members.

[0064] (vi) In the specific embodiment described above, an example was described in which the cap mounting portion 22 has the recess 222, but the cap CL may be mounted directly on a flat position without providing the recess 222. Furthermore, the cap CL may be mounted integrally with the portion on which it is mounted, or may be a separate portion. [Explanation of symbols]

[0065] 1 stage, 1a work surface, 2 Mounting member, 21 Container body storage section, 211 Fixed storage section, 212 Movable storage section, 22 Cap mounting section, 221 Cap receiving member, 222 Recessed section, 3 capper, 31 engaging claw, 331 claw portion, 4 pipette parts, 41 pipette tips, 5 Robot, 51 First arm, 51J First joint, 52 Second arm, 52J Second joint, 53 Third arm, 53J Third joint, 54 Fourth arm, 541 Base, 542 Holder, 542P Connecting pin, 6 Safety cabinet, 61 Housing, 62 Work table, 63 Exhaust module, 631 Air intake port, 632 Air intake filter, 633 Blower, 634 Exhaust filter, 635 Exhaust port, 636 Exhaust filter differential pressure gauge, 64 Measurement module, 641 Particle counter, 642 PAO inlet side measurement valve, 65 Sterilization module, 66 Lighting module, 67 Movement adjustment module, 671 Roller, 672 Adjuster, 673 Support leg, 68 Operation display unit, C container, C1 first bottle, C2 second bottle, C3 third bottle, C4 fourth bottle, CB container body, CL cap, CH container body holder, V Vertical direction, H Horizontal direction, CU Control unit

Claims

1. a plurality of containers each having a container body and a cap engaging the container body; a stage provided with a capper that performs an operation of engaging or separating the cap with the corresponding container body, a pipette member that performs an operation of suctioning or discharging liquid from the container body, a robot that selectively grasps and operates the container, and a cap placement unit that temporarily places each of the caps removed from each of the container bodies; a control unit capable of controlling the operation of the robot, the engaging operation and the separating operation of the capper, and the liquid suction operation and the liquid discharge operation of the pipette member; a safety cabinet that provides a sterile working environment, wherein the stage is disposed within the safety cabinet, so that the cap placement unit, the capper, the pipette member, and the robot are all located within the working environment; The cell recovery device, wherein the cap placement portion, the capper, and the pipette member are spaced apart from each other in a plan view and are arranged to surround the robot.

2. The cell recovery device according to claim 1, further comprising a cap state detector capable of detecting whether the container is covered with the cap.

3. The cell recovery device according to claim 2, further comprising a movable container unit that holds the container body so as to tap the container body and / or tilt the container body relative to the stage.

4. 4. The cell recovery device according to claim 3, wherein the tapping is a reciprocating vibration operation that reciprocates once or twice per second for a period of 10 seconds or more.

5. 5. The cell recovery device according to claim 4, wherein the container further comprises a container body holder for fixedly holding the container body, and the container body holder has a gripping portion that is gripped by the robot.

6. The cell recovery device described in claim 5, characterized in that the part of the capper that grips the cap has two or more inner diameter dimensions, making it possible to grip and open and close caps with different diameter size ranges.

7. 7. The cell recovery device according to claim 6, wherein the cap placement section has a cap receiving member that can be grasped by the robot and that receives the cap separated from the container body.

8. A cell recovery method using the cell recovery device according to any one of claims 1 to 7, In the work process controlled by the control unit, a culture medium discharging step of discharging a culture medium in a culture vessel, which is one of the vessels; a washing step of injecting a cell washing solution into the culture vessel and then discharging the cell washing solution; a detachment step of injecting a cell detachment solution into the culture vessel to detach cells in the culture vessel; a washing step of washing the cells in the culture vessel; a recovery step of placing the cells in the culture vessel into a cell recovery bottle, which is one type of the vessel; a suspension step in which the pipette member repeatedly performs the liquid suction operation and the liquid discharge operation on the cell collection bottle, thereby suspending the cells in the cell collection bottle.

9. The cell recovery method according to claim 8, further comprising an additional recovery step of injecting a cell washing solution into the culture vessel after the recovery step and before the suspension step, and then placing the remaining cells into the cell recovery bottle using the cell washing solution.

10. The cell recovery method described in claim 8 or 9, characterized in that the cell recovery device is equipped with a cap state detector that can detect whether the container is covered by the cap, and detection is performed by the cap state detector after all of the engagement or separation operations related to the cap.

11. 10. The cell recovery method according to claim 9, wherein the shaking operation of the culture vessel is performed via the robot in any of the washing step, the detachment step, the recovery step, and the additional recovery step.

12. A cell recovery method according to any one of claims 8 to 11, characterized in that at least one of all operations involving the robot includes an operation of moving the robot to a reset position.

13. a plurality of containers each having a cap that engages with a container body; a capper that performs an engagement or separation operation between the cap and the container body; a pipette member for sucking and discharging liquid into and from the container body; a robot that selectively grasps and operates a container from among the plurality of containers; a cap placement portion for placing each of the caps separated from each of the container bodies; a control unit capable of controlling the operation of the robot, the engaging operation and the separating operation of the capper, and the liquid suction operation and the liquid discharging operation of the pipette member, A cell recovery device, characterized in that the cap placement portion, the capper, and the pipette member are spaced apart from each other in a plan view and are arranged to surround the robot.

14. a stage provided with the capper, the pipette member, the robot, and the cap placement unit; 14. The cell recovery device according to claim 13, further comprising a safety cabinet that forms a sterile working environment therein and in which the stage is disposed.

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