CN113493738A

CN113493738A - Regenerative cell recovery device and cell recovery method

Info

Publication number: CN113493738A
Application number: CN202010193322.8A
Authority: CN
Inventors: 杉本昇平; 古野哲生
Original assignee: Rohto Pharmaceutical Co Ltd
Current assignee: Rohto Pharmaceutical Co Ltd
Priority date: 2020-03-18
Filing date: 2020-03-18
Publication date: 2021-10-12
Also published as: JPWO2021187611A1; WO2021187611A1

Abstract

The invention provides a regeneration cell recovery device and a cell recovery method. The regenerative cell recovery device includes a workbench and a control unit. The table is provided with a cap operation part, a pumping part, a robot, and a cap placement part. The lid operating member is used to operate the lids of different containers to realize the engagement and separation operations of the lids and the corresponding container bodies. The pumping part is used to perform suction and discharge operations on the container body of the container. The robot can selectively hold the container and drive the container to move, and the lid placement portion is used to temporarily store the lids removed from the container bodies respectively correspondingly. In this way, according to a predetermined program, the control unit can control the cap operation part, the pumping part and the robot to perform precise operation, thereby replacing manual operation, saving labor in the field of regenerative cell recovery, and preventing errors in the operation process.

Description

Regenerative cell recovery device and cell recovery method

Technical Field

The invention relates to the field of regenerative cell recovery, in particular to a regenerative cell recovery device and a cell recovery method using the regenerative cell recovery device.

Background

In the field of regenerative cell recovery, it is necessary to perform a recovery operation of cultured cells using various containers. In most existing cases, such recovery operations often require manual handling by an operator. However, since different containers need to be frequently replaced in the operation process and strict requirements are imposed on the operation timeliness, strict execution of each step and sterile environment during operation, the manual operation of the operator has the problems of troublesome operation, large workload, extremely high skill requirement, easy error occurrence in the operation process and the like.

Disclosure of Invention

The present invention has been made in view of the above-mentioned drawbacks of the prior art. An object of the present invention is to provide a regenerative cell recovery apparatus and a cell recovery method, which can save labor at least partially in the field of regenerative cell recovery and prevent errors in the operation process.

In order to achieve the above object, the present invention adopts the following technical solutions.

The present invention provides a regenerative cell recovery device including:

a plurality of containers, each of the containers comprising a container body and a lid that mates with the container body;

a table provided with a cap operating part for performing an engaging operation or a separating operation between the cap and the corresponding container body, an aspiration and injection part for performing an inhaling operation and a discharging operation on the container body, a robot capable of selectively gripping the container and driving it to move, and a cap placing part for respectively temporarily storing each cap detached from each container body correspondingly;

a control unit capable of controlling the movement of the robot, controlling the engagement operation and the disengagement operation of the cap operating member, and controlling the suction operation and the discharge operation of the suction member; and

the safety cabinet is used for providing an aseptic working environment, the workbench is arranged in the safety cabinet, so that the cover placing part, the cover operating part, the pumping and injecting part and the robot are all located in the working environment.

Preferably, the regenerative cell recovery device further includes a lid state detector capable of detecting whether or not the container is in a state of being covered with the lid.

More preferably, the regenerative cell recovery device further includes a movable receiving portion that holds the corresponding container body so as to enable a beating motion of the container body and/or a tilting motion of the container body with respect to the table.

More preferably, the striking motion is a reciprocating oscillating operation with a duration of 10 seconds or more and a rate of 1 to 2 times of reciprocation per second.

More preferably, the container further includes a container body holding portion that fixedly holds the container body, and the container body holding portion has a gripping portion that the robot grips.

More preferably, the cap operating member has two or more inner diameter dimensions at a portion for gripping the cap, so that the cap can be opened and closed by gripping the caps having different diameter dimension ranges.

More preferably, the cap seating part has a cap receiving member capable of being grasped by the robot for receiving the cap separated from the container body.

The present invention also provides a cell recovery method using the regenerative cell recovery device according to any one of the above aspects, wherein the operation process performed under the control of the control unit includes the following steps in order:

a medium discharging step of discharging the medium in the culture vessel in the vessel;

a cleaning step of pouring a cell cleaning solution into the culture container and then discharging the cell cleaning solution;

a peeling step of pouring a cell peeling solution into the culture container to peel the cells in the culture container;

an enzyme inactivation step of inactivating an enzyme in the cell-separating medium in the culture vessel;

a recovery step of discharging the cells in the culture vessel to a cell recovery flask in the vessel; and

and a suspending step of repeating the suction operation and the discharge operation with respect to the cell collection bottle by the suction member to suspend the cells therein.

Preferably, the cell recovery method further comprises a supplementary recovery step, after the recovery step and before the suspension step, in which a cell washing solution is poured into the culture vessel and then the remaining cells are poured into the cell recovery flask using the cell washing solution.

More preferably, the detection is performed by the lid state detector after all operations involving the engaging operation or the disengaging operation of the lid.

More preferably, in the washing step, the peeling step, the recovering step, and the replenishing and recovering step, the culture vessel is shaken by the robot.

More preferably, at least one of all operations of the robot is provided with an operation of moving the robot to an initial state.

By adopting the technical scheme, the invention provides the following regenerative cell recovery device and a cell recovery method adopting the regenerative cell recovery device. The regenerative cell recovery apparatus includes a table and a control unit. The workbench is provided with a cover operating part, a pumping part and a robot. And a cap placing portion. The cap operating means is used to operate the caps of different containers to perform engaging and disengaging operations of the caps with the corresponding container bodies. The pumping member is used for performing suction operation and discharge operation on the container body of the container. The robot can selectively hold the container and move the container, and the cap storage portion is used for respectively and temporarily storing the caps detached from the container bodies. With the above configuration, the control section can accurately control the engaging operation and the disengaging operation of the cap operating member, can accurately control the suction and discharge operations of the suction and discharge member in a precisely fixed amount, and can accurately control the robot to perform the movement, thereby replacing the manual operation, saving the labor in the regenerative cell recovery field, and preventing the occurrence of errors in the operation process, according to a predetermined program.

Drawings

FIGS. 1a and 1b are a front view and a top view, respectively, showing a regenerative cell recovery apparatus according to an embodiment of the present invention, in which a safety cabinet is omitted.

Fig. 2a and 2b show the structure of the first bottle, and fig. 2c shows a state where the first bottle is held by the container holding portion.

Fig. 3a and 3b show the structure of the second bottle, respectively, and fig. 3c shows a state where the second bottle is held by the container holding portion.

Fig. 4a shows the structure of the third bottle, and fig. 4b shows a state where the third bottle is held by the container holding portion.

Fig. 5a shows the structure of the fourth bottle, and fig. 5b shows a state where the fourth bottle is held by the container holding portion.

FIGS. 6a and 6b are a front view and a side view, respectively, illustrating a safety cabinet of a regenerative cell recovery apparatus according to an embodiment of the present invention, wherein FIG. 6a is an external view of the safety cabinet and FIG. 6b is a side view illustrating an internal portion thereof.

Description of the reference numerals

1 working surface of the table 1a

2 placing member 21 container body receiving portion 211 fixed receiving portion 212 movable receiving portion 22 cover placing portion 221 cover receiving member 222 concave portion

3-lid operating member 31 engaging claw 311 claw portion

4 pumping part 41 pumping head

5 robot 51 first arm part 51J first joint 52 second arm part 52J second joint 53 third arm part 53J third joint 54 fourth arm part 541 base part 542 holding part 542P connecting pin

6 safety cabinet 61 cabinet 62 cabinet 63 exhaust assembly 631 inlet 632 inlet filter 633 fan 634 exhaust filter 635 exhaust outlet 636 exhaust filter differential pressure meter 64 measurement assembly 641 particle counter 642 poly alpha olefin inlet side measurement valve 65 sterilization assembly 66 illumination assembly 67 movement adjustment assembly 671 roller 672 adjustment mechanism 673 leg 68 operation display portion

C container C1 first bottle C2 second bottle C3 third bottle C4 fourth bottle CB Container body CL Cap CH Container body retaining part

V the vertical direction H is horizontal.

Detailed Description

Exemplary embodiments of the present invention are described below with reference to the accompanying drawings. It should be understood that the detailed description is intended only to teach one skilled in the art how to practice the invention, and is not intended to be exhaustive or to limit the scope of the invention. In the present invention, the "vertical direction" refers to the direction of gravity of the earth (vertical direction in fig. 1 a), and the "horizontal direction" refers to the horizontal direction (horizontal direction in fig. 1 a) in the plane orthogonal to the vertical direction; the term "tilting motion" means a process in which the central axis of the container is tilted at a predetermined angle with respect to the vertical direction and then held immovably by the movable receiving portion, the term "striking motion" means a process in which the container is reciprocated within a predetermined range at a predetermined frequency in a predetermined posture so that the container can be oscillated reciprocally by the movable receiving portion, and the term "shaking" means a combined motion process in which the container is oscillated and shaken reciprocally within a predetermined range by a robot, and the central axis of the container may be in the vertical direction or in the horizontal direction or may not be in a fixed direction.

The structure of a regenerative cell recovery apparatus according to an embodiment of the present invention will be described below with reference to the drawings.

(according to an embodiment of the invention of the regenerative cell recovery device structure)

As shown in fig. 1a to 6b, the regenerative cell recovery apparatus according to an embodiment of the present invention includes a safety cabinet 6, a control part (not shown) provided outside the safety cabinet 6, and a plurality of containers C provided inside the safety cabinet 6, a table 1, a placement part 2, a lid manipulation part 3, a pumping part 4, and a robot 5.

In the present embodiment, the container C is used to contain a fluid such as a culture medium or a cell solution for recovering regenerative cells, and can be held by the robot 5 and moved to various positions. As shown in fig. 2a, 2b, 3a, 3b, 4a and 5a, each container C includes a container body CB forming a receiving space and a corresponding lid CL for closing an opening of the container body CB, and a container body holding portion CH fixedly holding the container body CB. Specifically, in the present embodiment, the container C is divided into a first bottle C1, a second bottle C2, a third bottle C3, and a fourth bottle C4.

The first bottle C1 is a bottle having a container body CB with a square cross section and a volume of 125ml for containing a cell washing solution such as PBS (phosphate buffered saline) or a cell peeling solution such as pancreatin. Before the operation of the regenerative cell recovery apparatus according to the present invention is started, the first bottles C1 are stored in the storage positions formed by the movable storage portions 212 in a one-to-one correspondence, and the movable storage portions 212 are capable of rotating about an axis in the up-down direction V and linearly displacing in the up-down direction V. The container body holding portion CH of the first bottle C1 is provided with a gripping portion corresponding to the container holding portion 54 of the robot 5.

The second bottle C2 is a bottle having a container body CB of which the cross section is circular and the volume is 500ml, the lower half of the container body CB of the second bottle C2 is formed in a tapered shape toward the bottom surface, the second bottle C2 may be a culture container, such as a multi-layer culture container, for cell recovery, and the second bottle C2 may further include other structures facilitating the concentration, flow, distribution and recovery of a cell solution. Before the operation of the apparatus for collecting regenerative cells according to the present invention is started, the second bottle C2 may be stored in the movable storage section 212 or the fixed storage section 211. The container body holding portion CH of the second bottle C2 is provided with a gripping portion corresponding to the container holding portion 54 of the robot 5.

The third bottle C3 is a bottle having a rectangular cross section of the container body CB. Before the operation of the regenerative cell recovery apparatus according to the present invention is started, the third bottles C3 are stored in the storage positions of the fixed storage portions 211 in a one-to-one correspondence, and are fixed to the table 1. The third flask, C3, served as a culture container for the media and the growing cells that will eventually be collected in the second flask, C2. The container body holding portion CH of the third bottle C3 is provided with a gripping portion corresponding to the container holding portion 54 of the robot 5.

The fourth bottle C4 is a bottle having a container body CB whose cross section is square and whose volume is 2000 ml. Before the operation of the regenerative cell recovery apparatus according to the present invention is started, the fourth bottles C4 are stored in the storage positions of the fixed storage portions 211 in a one-to-one correspondence, and are fixed to the table 1. The fourth bottle C4 is a bottle for containing the discharged medium, and all the medium discharged from the third bottle C3 is collected in the fourth bottle C4.

The container body holding portions CH of all of the four bottles C1, C2, C3, and C4 may be dedicated resin holders, and the container body holding portions CH may be manually bound with silicone rubber. Thus, even if the bottles are inverted, the bottles cannot fall off; in addition, since each of the bottles C1, C2, C3, C4 is made of plastic, when the container body CB is directly grasped by the robot 5, there are cases where it is flexed to make the capping operation difficult and the bottles are liable to fall, but the presence of the container body holding portion CH makes it possible to avoid the occurrence of these cases.

In the present embodiment, as shown in fig. 1a and 1b, the table 1 is placed on a work table in the safety cabinet 6. The table 1 includes a work surface 1a extending in a horizontal direction H, and a placement member 2, a lid operating member 3, a drawing member 4, and a robot 5 are provided on the work surface 1 a.

Specifically, in the present embodiment, the placement member 2 is used to place the container body CB and the lid CL of the container C, respectively. The placement member 2 includes a container body receiving portion 21 and a lid placement portion 22, and the container body receiving portion 21 and the lid placement portion 22 form a placement position corresponding to the container body CB and the lid CL of the container C, respectively, one to one, and a temporary storage position for temporarily storing the container C and a tilting/rocking position for causing the container body CB of the container C to perform tilting movement (normally, the lid CL is not engaged)/reciprocating movement (normally, the lid CL is engaged).

Further, the container body accommodating portions 21 are for accommodating the container bodies CB corresponding to the containers C and have an accommodating space matching the sectional shape of the container bodies CB, and a plurality of container body accommodating portions 21 are distributed on the table 1 in such a manner as to surround the robot 5, so that the robot 5 can operate the containers C in the container body accommodating portions 21 as conveniently as possible. The container body housing portion 21 includes a fixed housing portion 211 and a movable housing portion 212 depending on whether or not the container body housing portion 21 can make relative movement.

The fixed receiving portion 211 holds the corresponding container body CB so that the corresponding container body CB is fixed to the operating surface 1a, and in the present embodiment, the fixed receiving portion 211 makes the center axis of the corresponding container body CB perpendicular to the operating surface 1a, that is, the center axis of the container body CB extends in the vertical direction V. In this way, the opening of the container body CB accommodated in the fixed accommodating portion 211 is always directed upward in the vertical direction V. The fixed receiving portion 211 forms the above-mentioned placing position and temporary storage position.

The movable receiving portion 212 holds the corresponding container body CB so that the corresponding container body CB can rotate with respect to the operating surface 1 a. In the present embodiment, the movable receiving portion 212 for receiving the first bottle C1 is rotatable about an axis extending in the up-down direction V and linearly displaceable in the up-down direction V, so that the first bottle C1 can be at different positions in the predetermined up-down direction V and different positions in the horizontal direction H. If necessary, containers C other than the first bottle C1 may be provided in the movable storage section 212 so that each container C can be moved to an advantageous position where the robot 5 operates each container C. The movable housing portion 212 is also formed at the above-described tilting/rocking position so that the container C moved to the tilting/rocking position by the robot 5 can perform a desired tilting movement and/or reciprocating movement in the movable housing portion 212. The movable housing portion 212 for forming the above-described tilting/rocking position houses only a specific container C when necessary, so that the central axis of the container body CB of the container C can be tilted with respect to the horizontal work surface 1a and/or the container C is caused to perform a reciprocating motion within a predetermined range, promoting uniform mixing and distribution of the solution in the container C.

Further, the cap housing portion 22 is used to house the cap CL screwed off from the container C, thereby forming a housing position for housing the cap CL. In the present embodiment, the cover placing portion 22 has a cover receiving member 221 that can be grasped by the robot, and the cover receiving member 221 is formed with a plurality of concave portions 222 that match the shape of each cover CL so that the concave portions 222 can accommodate the corresponding covers CL, one concave portion 222 corresponding to the placing position of one cover CL. The cover receiving member 221 can be moved by the robot 5 from the table 1 to the cover operating part 3 and align the corresponding recess 222 on the cover receiving member 221 with the cover CL screwed down by the cover operating part 3, thereby placing the cover CL in the recess 222. In addition, the cap receiving member 221 can be moved by the robot 5 from the table 1 to the cap operating part 3 and align the cap CL placed on the cap receiving member 221 with the cap operating part 3, thereby engaging the cap operating part 3 with the cap CL and then screwing the cap CL onto the corresponding container body CB.

In the present embodiment, the cap operating means 3 is used to operate the caps CL of different containers C to perform the engaging and disengaging operations of the caps CL with the corresponding container bodies CB. The cap operating member 3 includes engaging claws 31 that are engageable with the cap CL of the container C and are rotatable bidirectionally about the up-down direction V, and the engaging claws 31 include three claw portions 311 that are evenly distributed in the circumferential direction of the cap CL so that the engaging claws 31 can screw/unscrew the cap CL of the container C. After the engagement claws 31 are engaged with the lid CL of the container C mounted to the container body CB from the outside, the rotation of the engagement claws 31 toward one direction can unscrew the lid CL from the container body CB, thereby separating the lid CL from the container body CB; on the contrary, the engagement claws 31 can engage the lid CL and the container body CB together, thereby closing the opening of the container body CB.

The lid operating unit 3 further includes a lid state detector (not shown) capable of detecting whether or not the container C is covered with the lid CL, and the lid state detector may be an optical sensor. In the cell recovery method described below, after all operations involving the engaging operation or the separating operation of the lid CL, the lid state detector detects the operations to determine whether or not the lid CL is securely engaged with and separated from the container body CB. When the control part detects that the work is not in the expected state, the control part controls the buzzer to give an alarm and suspends the work.

In addition, in the lid operating member 3, a clamping assembly for clamping the container body CB of the container C may be provided, the clamping assembly including two clamping portions that can approach and separate from each other, so that the container body CB of the container C can be fixedly held by the two clamping portions, so that the container body CB is relatively fixed. In this way, in the process of engaging and disengaging the lid CL with and from the container body CB by the operation of the lid operating member 3 on the lid CL of the container C, the container body CB is not freely rotated in accordance with the operation of the engaging claws 31 of the lid operating member 3. Further, the inner peripheral wall of the engagement claw 31 may be formed with engagement portions having two or more inner diameter sizes, so that the engagement claw 31 can be engaged with the caps CL having different diameter ranges, thereby being able to grip the caps CL having different diameter size ranges. For example, when the engaging portions of the inner peripheral wall of the engaging claw 31 are formed in the inner diameter sizes of 5cm and 7cm, the cap CL of 5cm to 6cm in diameter can be engaged and also the cap CL of 7cm to 8cm in diameter can be engaged. Of course, any design with different inner diameter dimensions may be adopted, and the specific applicable dimension range may be adjusted according to specific requirements.

In this embodiment, the drawing member 4 has a pipette (50ml), a silicone tube, and a glass syringe assembled together. The pipette is used for directly sucking and discharging the solution in the container body CB by extending into the container body CB. The silicone tube is located between the pipette and the glass syringe and has a certain flexibility and functions to hermetically connect the pipette and the glass syringe so that the movement of the pipette is not hindered by the inability of the connecting portion to deform when the pipette is moved in the up-down direction V. Both the pipette and the silicon tube are disposable. The glass syringe can act as a piston to power the aspiration and discharge operations. In this way, the drawing and injecting section 4 serves to perform a suspension treatment (dispersion/homogenization/suspension of cells) of the cells in the cell solution recovered into the second bottle C2. The pumping means 4 sucks the solution from the container body CB of the container C at different fixed amounts (sucking operation) or discharges the solution to the container body CB of the container C (discharging operation) to perform the above-mentioned floating treatment.

In the present embodiment, the mechanical structure of the robot 5 has six degrees of freedom, and moves the container body CB and the lid receiving member 221 of the container C. The robot 5 includes a plurality of

arm portions

51, 52, 53, 54 connected in sequence, wherein a container holding portion 542 as an end portion of the fourth arm portion 54 of the terminal arm is capable of holding the container C in at least two relative attitudes (corresponding to the first holding mode and the second holding mode) with respect to the container C, and when the container holding portion 542 holds the container C in the first holding mode, the holding portion 542 of the fourth arm portion 54 is angled at a first angle with respect to the central axis of the container body CB of the container C, and when the container holding portion 542 holds the container C in the second holding mode, the angle formed by the holding portion 542 of the fourth arm portion 54 and the central axis of the container body CB of the container C is a second angle, and the difference between the first angle and the second angle may be any value greater than 0 degrees and less than 180 degrees, so that the container C is brought into a predetermined movement (linear displacement movement and shaking). The fourth arm portion 54 has a base portion 541 extending in the first direction and a holding portion 542 extending in the second direction orthogonal to the first direction, the base portion 541 and the holding portion 542 are fixedly connected together, the base portion 541 is connected to a third arm portion 53 described below, and the holding portion 542 is used to hold the container C. The holding portion 542 can hold the container C in such a manner that the center axis of the container C coincides with the first direction or with the second direction, thereby achieving holding of the container C via the container body holding portion CH in the first holding mode and the second holding mode, respectively, when the difference between the first angle and the second angle is 90 degrees. In the case where the central axis of the container C coincides with the first direction (as shown, for example, in fig. 2C, 3C, 4b and 5 b), the robot 5 mainly makes the container C perform a linear translation in an attitude with the opening facing upwards, that is, for moving the container C between different positions. In the case where the centerline axis of the container C coincides with the first direction and with the second direction, the robot 5 can tilt the opening of the container C at an arbitrary angle with respect to the up-down direction V to pour the solution in the container body CB or can shake the container C while holding the container C to mix the solution in the container C uniformly. In this way, the solution in the container C is poured by two opposite postures, and the solution in the container C can be poured to more thoroughly prevent excessive solution from remaining similarly to the effect of the human body to positively hold and reversely hold the container C; shaking the container C with two relative postures enables the solution inside the container C to be mixed more uniformly. In the present embodiment, the holding portion 542 includes a first holding portion and a second holding portion that are located at different positions, and the first holding portion and the second holding portion are capable of holding the same portion of the container C, and are in the first holding mode when the first holding portion is held and in the second holding mode when the second holding portion is held. In addition, the holding portion 542 includes a connecting pin 542P for connecting with the container C so as to hold the container C by clamping via the connecting pin 542P, and the connecting pin 542P is engageable with a predetermined held hole of the container C so that the holding portion 542 and the container C are relatively fixed. In the present embodiment, two sets of the connecting pins 542P are provided, two connecting pins 542P are provided for each set of the connecting pins 542P, and one set of the connecting pins 542P and the other set of the connecting pins 542P are provided so as to protrude in opposite directions.

Further, the plurality of

arms

51, 52, 53 of the robot 5 include a first arm 51 having one end provided on the table 1 of the regenerative cell recovery device, a second arm 52 connected to the first arm 51, and a third arm 53 connected to the second arm 52, in addition to the fourth arm 54. The first arm 51 extends in the vertical direction V and is rotatable about the vertical direction V, the second arm 52 extends linearly, one end of the second arm 52 is connected to the other end of the first arm 51 via a first joint 51J, the second arm 52 has at least one rotational degree of freedom with respect to the first arm 51 by the first joint 51J, the third arm 53 extends linearly, one end of the third arm 53 is connected to the other end of the second arm 52 via a second joint 52J, the third arm 53 has at least one rotational degree of freedom with respect to the second arm 52 by the second joint 52J, the fourth arm 54 is connected to the third arm 53 by the third joint 53J, and the fourth arm 54 has at least one rotational degree of freedom with respect to the third arm 53 by the third joint 53J. More specifically, the first joint 51J is provided with a first shaft extending in the horizontal direction H, the second arm portion 52 is rotatable about the first shaft, the second joint 52J is provided with a second shaft extending in the horizontal direction H, the third arm portion 53 is rotatable about the second shaft, the third joint 53J is provided with a third shaft extending in the horizontal direction H, the fourth arm portion 54 is rotatable about the third shaft, and both the third arm portion 53 and the fourth arm portion 54 are rotatable about respective central axes. In this way, the robot 5 can realize six degrees of freedom of movement.

By adopting the above configuration, the robot 5 has a six-degree-of-freedom mechanical structure, and can hold other components in various required postures, thereby completing operations such as linear displacement, rotation, tilting, and shaking of the container C. Specifically, the robot 5 can move each container C to the lid operating part 3, can align each concave portion 222 of the lid receiving member 221 with the unscrewed lid CL, can move each container C to the placement position, the temporary storage position, the tilting position, and the like, and can perform various required works smoothly.

In the present embodiment, the control unit can control the robot 5 to perform various operations on the container C according to a predetermined program, control the engagement operation and the disengagement operation of the cap operating member 3, and control the drawing member 4 to selectively perform the suction operation and the discharge operation. Although not explicitly described in the present embodiment, the control section may include a plurality of corresponding control units for controlling the operations of the respective components so as to perform the various operations described above according to a predetermined program.

In the present embodiment, as shown in fig. 6a and 6b, the safety cabinet 6 is used to provide a sterile work environment, and the work table 12 may be provided on a detachable work table 62 in the safety cabinet 6 and may be the same as the work table 62. The safety cabinet 6 includes a cabinet body 61, an exhaust assembly 63, a measurement assembly 64, a sterilization assembly 65, a lighting assembly 66, and a movement adjustment assembly 67.

Specifically, in the present embodiment, the cabinet 61 has a cubic shape and the inside forms a sterile working environment, and the work table 62 is mounted inside the cabinet 61 and arranged in a horizontal manner.

Further, in the present embodiment, the exhaust assembly 63 serves to communicate the inside of the cabinet 61 with the outside. The exhaust assembly 63 includes an intake port 631 and an exhaust port 635 provided to the cabinet 61, intake filter sheets 632 (two in the present embodiment) provided to the cabinet 61 at the intake port 631, and exhaust filter sheets 634 (one in the present embodiment) provided to the cabinet 61 at the exhaust port 635, so as to filter dust, particles, and the like during gas exchange between the inside and the outside of the cabinet 61. The exhaust assembly 63 further includes a fan 633 provided inside the exhaust opening 635, the fan 633 for facilitating air inside the cabinet 61 to be exhausted to the outside. In addition, the exhaust assembly 63 further includes an exhaust filter differential pressure gauge 636 for monitoring the differential pressure of the exhaust filter 634.

Further, in the present embodiment, the measurement assembly 64 is used to measure a predetermined parameter of the interior of the cabinet 61. More specifically, the measurement assembly 64 includes a particle counter 641 for measuring the concentration of dust and particulates within the cabinet 61 and a polyalphaolefin inlet side measurement valve 642 for measuring the polyalphaolefin input, thereby providing the user with the desired parameter information.

Further, in the present embodiment, the sterilization unit 65 is provided in the cabinet 61 and is configured to sterilize the inside of the cabinet 61. Specifically, the sterilizing assembly 65 includes two sterilizing lamps (15 w) disposed at the side wall of the cabinet 61, so that the inner space of the cabinet 61 can be effectively sterilized.

Further, in the present embodiment, the illumination assembly 66 is used to illuminate the inside of the cabinet 61. Specifically, the lighting assembly 66 includes three LEDs disposed at the top of the interior space of the cabinet 61.

Further, in the present embodiment, a movement adjusting assembly 67 is provided at a lower portion of the cabinet 61 for moving the cabinet 61 and adjusting an angle between the cabinet 61 and the ground. Specifically, the movement adjustment assembly 67 includes a plurality of rollers 671, an adjustment mechanism 672, and a leg 673. The rollers 671 facilitate travel over the ground as the cabinet 61 moves. The adjustment mechanism 672 is used to support the cabinet 61 on the one hand and to adjust the angle between the cabinet 61 and the ground on the other hand. The legs 673 can provide support for the cabinet 61 when lowered.

Further, in the present embodiment, the operation display unit 68 is provided on the outer wall of the cabinet 61, and is used to provide parameter display inside the cabinet 61, input an operation command for controlling the operation of each component of the safety cabinet 6, and the like.

The present invention is not limited to the above-described embodiments, and various modifications and variations can be made without departing from the spirit and scope of the present invention.

(example of operation of the apparatus for collecting regenerative cells according to one embodiment of the present invention)

In the operation of the regenerative cell recovery apparatus according to an embodiment of the present invention, the method includes the steps of:

a medium discharging step of discharging the medium in the third bottle C3 as a culture container into the fourth bottle C4 by the robot 5 so that most of the medium in the third bottle C3 is discharged in the step;

a washing step of pouring the cell washing solution stored in the first bottle C1 into the third bottle C3 by the robot 5 so that the residual medium can be completely washed by the cell washing solution, and then discharging the cell washing solution into the fourth bottle C4, wherein the third bottle C3 is tilted to one side by the movable storage part 212 to facilitate the pouring work when the cell washing solution is poured into the third bottle C3;

a peeling step of pouring a cell peeling solution stored in the first bottle C1 into the third bottle C3 by the robot 5 to peel off the cells in the third bottle C3, and when pouring the cell peeling solution into the third bottle C3, tilting the third bottle C3 to one side by the movable containing part 212 to facilitate the pouring operation;

an enzyme inactivation step of inactivating the enzyme in the cell stripping solution in the third vial C3 by appropriately heating the third vial C3 with a heating device not shown in the figure, for example, and the cells in the third vial C3 are not affected by the heating;

a recovery step of discharging the cell separation solution and the cells in the third vial C3 to the second vial C2 as a cell recovery vial by using the robot 5; and

and a suspension step of suspending the cells in the second vial C2 using the aspiration and injection member 4.

Further, the cell recovery method further comprises a supplementary recovery step after the recovery step and before the suspension step. In the replenishment recovery step, the cell washing solution is poured into the third bottle C3, and then the residual cells are poured into the second bottle C2 using the cell washing solution, so that the cells remaining in the third bottle C3 can be recovered into the second bottle C2 as sufficiently as possible.

In the cleaning step, the peeling step, the recovering step, and the replenishing and recovering step, the third bottle C3 is shaken by the robot 5. The shaking operation was performed in a state where the thickness direction of the third bottle C3 was substantially in the up-down direction, and the culture container was turned upside down at least once during the shaking operation, which did not affect the state where the thickness direction was substantially in the up-down direction. In the recovery step and the supplementary recovery step, the third bottle C3 may be placed in the movable storage 212 during the shaking operation by the robot 5, and the third bottle C3 may be displaced back and forth by the movable storage 212, for example, by a back and forth shaking operation in which the duration is 10 seconds or more and the speed is 1 to 2 times per second, and the shaking operation may be stopped immediately after the back and forth acceleration to simulate an effect similar to a beat, thereby allowing the cells in the third bottle C3 to be sufficiently taken into the solution and the solution in the third bottle C3 to be sufficiently mixed, thereby achieving sufficient recovery.

The operation of pouring the liquid in the above steps is performed by the pouring operation of the robot 5, and the liquid in the container C is discharged in the pouring operation 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 completely discharged as much as possible. In all the steps described above, it is preferable that an operation of moving the robot 5 to the initial state is provided in the process of connecting all the operation operations of the robot 5. In this way, it is convenient to set an initial state in which the robot 5 performs the next action and it is easy to remove a trouble when the robot 5 malfunctions. After the fault is cleared, the next action is simply to continue downward from the initial state set in the action. Here, the operation of moving the robot 5 to the initial state is provided in all the operation links of the robot 5, but the operation of moving the robot 5 to the initial state may be provided only in a part of the operation operations of the robot 5. In addition, during most of the time period of the suspension step, the suction and injection member 4 is extended below the liquid level of the liquid in the second bottle C2 to perform suction and discharge operations, thereby performing the suspension process; at the final stage of the suspension step, the pumping means 4 is moved away from the liquid surface of the liquid in the second bottle C2, but the pumping operation is repeated in the air, whereby the cells remaining in the pumping means 4 can be completely pumped into the second bottle C2.

In summary, the present invention provides a novel regenerative cell recovery device, which is not limited to the above-mentioned embodiments. In addition, supplementary explanation is made as follows.

(i) Although it is described in the above embodiment that the working surface 1a of the table 1 is horizontal, the present invention is not limited thereto, and the working surface 1a may be set at a predetermined angle with respect to the horizontal plane as needed.

(ii) Although the structure of the safety cabinet 6 of the regenerative cell recovery apparatus according to the present invention is described in the above embodiment, the present invention is not limited thereto, the safety cabinet 6 may further have a required power outlet and power connection lines, and the cabinet body 61 may include a detachable structure for maintenance and a transparent structure for viewing the inner space.

(iii) Although it has been described in the above embodiment that the robot 5 of the regenerative cell recovery apparatus according to the present invention has six degrees of freedom, the present invention is not limited thereto. The robot 5 may also be a redundant robot with seven degrees of freedom.

(iv) Although the above embodiment describes an example in which the cap CL is gripped by the engagement claw 31, the means for gripping the cap CL by the cap operating member 3 is not limited to the claw type, and may be another type of gripping portion, and the other gripping portion may be provided with a plurality of inner diameters that can accommodate a plurality of gripping size ranges.

(v) Although the above embodiment describes an example in which the movable housing portion 212 can perform both the reciprocating oscillating motion and the tilting motion in the simulated impact, the movable housing portion 212 may perform various motions by different members.

(vi) Although the example in which the cover placing portion 22 has the concave portion 222 has been described in the above embodiment, the cover CL may be placed directly at a flat position without providing the concave portion 222. The portions for placing the respective covers CL may be connected or separated.

Claims

1. A regenerative cell recovery device, wherein the regenerative cell recovery device comprises:

a plurality of containers (C), each of said containers (C) comprising a container body (CB) and a lid (CL) cooperating with said container body (CB);

A workbench (1) provided with a cap operating part (3), a drawing part (4), a robot (5) and a cap placing part (22), the cap operating part (3) The cap (CL) is used for joining or separating the corresponding container body (CB), and the pumping member (4) is used for the suction operation of the container body (CB). and spit out operation, the robot (5) can selectively hold the container (C) and drive it to move, and the cover placing part (22) is used to temporarily store the containers (C) respectively correspondingly and temporarily. CB) each said cover (CL) removed;

A control unit capable of controlling movement of the robot (5), controlling the joining operation and the separating operation of the lid operating member (3), and controlling the operation of the pumping member (4) the suction operation and the said discharge operation; and

A safety cabinet (6), the safety cabinet (6) is used to provide a sterile working environment, and the workbench (1) is arranged in the safety cabinet (6) so that the cover placement portion (22) , the cover operating part (3), the pumping part (4) and the robot (5) are all located in the working environment.

2. The regenerative cell recovery device according to claim 1, wherein

The regenerative cell recovery apparatus further includes a lid state detector capable of detecting whether or not the container (C) is in a state where the lid (CL) is covered.

3. The regenerative cell recovery device according to claim 2, further comprising a movable accommodating part (212), wherein the movable accommodating part (212) is capable of hitting the container body (CB) The corresponding container body (CB) is held by moving and/or tilting the container body (CB) relative to the table (1).

4 . The regenerative cell recovery device according to claim 3 , wherein the hitting motion is a reciprocating oscillating operation with a duration of more than 10 seconds and a rate of 1-2 times per second. 5 .

5 . The regenerative cell recovery device according to claim 4 , wherein the container further comprises a container body holding part (CH) for fixedly holding the container body (CB), the container body holding part (CH) having A gripping part to be gripped by the robot.

6. The regenerative cell recovery device according to claim 5, wherein the portion of the lid operating member (3) for holding the lid (CL) has two or more inner diameters so that the diameter can be grasped The lids (CL) with different ranges can be opened and closed.

7. The regenerative cell recovery device according to claim 6, characterized in that the lid placement part (22) has a lid receiving member (221) capable of being grasped by the robot for receiving the container from the container The cover (CL) separated from the body (CB).

8. A cell recovery method using the regenerative cell recovery device according to any one of claims 1 to 7, characterized in that the work process under the control of the control unit comprises the following steps in sequence:

The culture medium discharge step is to discharge the culture medium in the culture container in the container (C);

In the washing step, the cell washing liquid is poured into the culture vessel, and then the cell washing liquid is discharged;

In the stripping step, a cell stripping solution is poured into the culture container to strip the cells in the culture container;

Enzyme inactivation step, inactivating the enzyme in the cell stripping solution in the culture vessel;

a recovery step of discharging the cells in the culture vessel to a cell recovery flask in the vessel (C); and

In the suspending step, the suction operation and the discharge operation are repeatedly performed on the cell recovery bottle by the pumping part (4), thereby suspending the cells therein.

9. The cell recovery method according to claim 8, wherein the cell recovery method further comprises a supplementary recovery step after the recovery step and before the suspending step, and in the supplementary recovery step, in the The cell washing solution is poured into the culture vessel, and the residual cells are then poured into the cell recovery flask by using the cell washing solution.

10. The cell recovery method according to claim 8 or 9, characterized in that, using the regenerative cell recovery device according to any one of claims 2 to 7, in all parts involving the cover (CL) After the operation of the joining operation or the separation operation, the lid state detector is used for detection.

11 . The cell recovery method according to claim 9 , wherein, in the cleaning step, the stripping step, the recovery step and the supplementary recovery step, the robot is used for the culturing vessel. 12 . Perform a shaking operation.

12. The cell recovery method according to any one of claims 8 to 11, characterized in that, in at least one step of all operations of the robot (5), a device is provided to move the robot (5) to a Action in the initial state.