CN115279881A - Cell recovery device - Google Patents

Abstract

The cell recovery device of the present invention has: an outer structure part, a shutter part opened and closed by moving up and down on the front part, and the interior can be made into a clean environment; a manipulator; a culture container setting table, on which the culture container for culturing cells is placed; a reagent container A stage is set for placing reagent containers; a cell recovery container is set up for placement of a cell recovery container, wherein the cell recovery container recovers the cells taken out from the culture container, and each stage can be held by the manipulator within the moving range. The container is provided inside the outer structure, the culture container table is provided near the left and right center of the outer structure and between the shutter and the manipulator, the reagent container The placement platform and the cell recovery container placement platform are arranged on the left and right sides and rear of the culture container placement platform.

Description

cell recovery device

Cross-reference of related applications

This application claims priority based on Japanese Patent Application No. 2020-036640, which is incorporated by reference in the description of this application specification

technical field

The invention relates to a cell recovery device for automatically recovering cells.

Background technique

In recent years, the cultivation of tissues, cells, fertilized eggs, etc. from various parts of the human body, and the use of the cultured cells in regenerative medicine have been put into practical use. One type of cell culture as adherent cells is static culture (monolayer culture), in which cells are sown into culture vessels such as culture dishes and culture bottles, and then placed in an incubator to allow the cells to attach to the culture vessel and Culture was performed in a monolayer state. Adherent cells are subcultured by taking out the culture medium from the culture vessel, washing the cell surface with PBS, etc., detaching the cells from the culture vessel with enzymes (such as trypsin), and then transferring a desired amount of cells to It was carried out with a new culture container filled with culture medium.

In the past, the subculture of cells was performed manually by humans. Especially for cells used in regenerative medicine, etc., it is necessary to safely cultivate high-quality cells, so cell culture specialists are required to perform cell subculture. However, it takes time to train a cell culture specialist, and the operational burden on the cell culture specialist is also heavy.

On the other hand, there is an apparatus (for example, described in JP 2008-54690 A) which can automate cell culture. However, such an apparatus automates medium exchange and subculture as a series of operations, so the apparatus is large and its installation location is limited.

In addition, although it is hoped that cells can be cultured and used in patients in hospitals and clinics, it is difficult to introduce automated devices in reality due to restrictions on installation locations in hospitals and clinics. Therefore, it is desired to develop a small replacement device that can be installed in hospitals, clinics, and the like.

prior art literature

patent documents

Patent Document 1: Japanese Patent Laid-Open No. 2008-54690.

Contents of the invention

The problem to be solved by the invention

Therefore, in view of the above circumstances, the object of the present invention is to provide a compact cell recovery device which is exclusively used for cell recovery for cell subculture.

solutions to problems

The present invention is a cell recovery device, comprising: an outer structure part, which has a gate part that can be opened and closed by moving up and down on the front part, and can make the inside a clean environment; a robot arm that is installed inside the outer structure part; , for placing the culture container for culturing cells; the reagent container is placed on a table, for placing the reagent container with reagents, wherein the reagent is used to take out cells from the culture container; the cell recovery container is placed on a table, for the cell recovery container to be placed, wherein the The cell recovery container recovers the cells taken out from the culture container, and each stage is installed inside the outer structure in such a manner that the manipulator can hold each container within the moving range of the manipulator. The container stand is installed near the left and right center of the outer structure part and between the gate part and the manipulator, and the reagent container stand and the cell recovery container stand are arranged on the left and right sides of the culture container stand. side and rear.

In addition, the reagent container table can also move back and forth inside the outer structure part.

In addition, it may also be configured such that a front air intake portion with an upper opening is provided in front of the outer structure portion and behind the shutter portion, and the culture container stand is provided on a front panel with air permeability in the vertical direction, so that A part of the front panel is provided to cover a part of the front air intake part from above.

Description of drawings

FIG. 1 is a front view schematically showing the structure of a cell recovery device according to one embodiment of the present invention.

FIG. 2 is a plan view schematically showing the internal structure of the cell recovery device, omitting the illustration of the top of the external structure.

Fig. 3 is a right side view showing schematically the internal structure of the cell recovery device, omitting the illustration of the side part of the external structure part.

Fig. 4A is a front view showing a hand (or grip) in a manipulator of the cell recovery apparatus.

Fig. 4B is a right side view showing a hand (or grip) in the manipulator of the cell recovery apparatus.

Fig. 4C is a view taken along the line c-c of Fig. 4B showing the hand (or holding part) in the manipulator of the cell recovery device.

Fig. 5A is a front view and a right side perspective view showing the left and right side parts of the hand (or grip part).

Fig. 5B is a front view showing the left and right sides of the hand (or grip).

Fig. 6A is a rear view showing the left and right sides of the hand (or grip).

Fig. 6B is a plan view showing the left and right sides of the hand (or grip).

Fig. 7A is a bottom view showing the left and right sides of the hand (or grip).

Fig. 7B is a right side view showing the left and right sides of the hand (or grip).

Fig. 8A is a left side view showing the left and right sides of the hand (or grip).

Fig. 8B is a perspective view showing the back and left sides of the left and right sides of the hand (or grip).

Fig. 9A is a front view showing both a culture vessel and a holding unit used in the cell recovery device.

Fig. 9B is a plan view showing both the culture container and the holding unit used in the cell recovery device with the cover removed.

Fig. 10A is a front view showing a holding part for a culture container used in the cell recovery device.

Fig. 10B is a plan view showing a holding portion for a culture container used in the cell recovery apparatus.

Fig. 10C is a bottom view showing a holding part for a culture container used in the cell recovery device.

Fig. 10D is a left side view showing a holding part for a culture container used in the cell recovery device.

Fig. 11A is a front view showing both a cell recovery container and a holding unit used in the cell recovery device.

Fig. 11B is a plan view of the holding unit alone.

Fig. 12A is a front view showing both a reagent container and a holding unit used in the cell recovery device.

Fig. 12B is a plan view showing both a reagent container and a holding unit used in the cell recovery device.

Fig. 12C is a rear view showing both the reagent container and the holder used in the cell recovery device.

Fig. 13A is a front view showing both a culture medium recovery container and a holding unit used in the cell recovery device.

Fig. 13B is a plan view of the holding unit alone.

Fig. 14A shows the first state in which the manipulator of the cell recovery device grips the culture container to which the holder is attached.

Fig. 14B shows the second state in which the manipulator of the cell recovery apparatus grips the culture container to which the holder is attached.

Fig. 15 is a vertical diagram showing the relationship between the rotating grip part and the cover (an example of a large cover shown by a solid line and an example of a small cover shown by a two-dot chain line) in the cover opening and closing device of the cell recovery device. cutaway view.

Detailed ways

Hereinafter, a cell recovery device 1 according to an embodiment of the present invention will be described. Furthermore, in the following representations regarding directions, "up and down" corresponds to the directions shown in FIG. 1 . In addition, regarding "front and rear", the lower side shown in FIG. 2 (the side where the gate part 102 shown in FIG. 3 is located) is regarded as the front, and the upper part shown in FIG. as the rear. In addition, about "left and right", it corresponds to the direction shown in FIG.1 and FIG.2.

The cell recovery device 1 of the present embodiment is a device for recovering cultured cells only. Therefore, recovery of a large number of cultured cells is basically not conceived unlike a device that performs a series of cell cultures. However, the technical scope of the present invention does not exclude the recovery of cells cultured in large quantities.

The cell recovery device 1 of this embodiment is structured as shown in FIGS. 1 to 3 , and mainly includes an external structure 10 , a manipulator 11 , a culture container stand 121 , a reagent container stand 13 , and a cell recovery container stand 122 . In addition, in addition to these, it also has a culture medium recovery container setting platform 123, a cover opening and closing device 14, a cover sensor 15, a cover setting platform 16, a tilting platform 17, a vibration table 18, a pipette device 19, a temporary storage platform 20, and an operating system. Panel 21. In the cell recovery apparatus 1, the movement of physically active parts (manipulator 11, reagent container setting table 13, cover opening and closing device 14, tilt table 17, vibration table 18, pipette device 19) is controlled by a control unit (not shown). ) to control.

The outer structure part 10 is box-shaped and has a space inside, and various structures necessary for cell recovery are disposed in the space. With respect to the table portion 101 in the interior of the outer structure portion 10 , which has a rectangular plane facing upward and extending left and right, some of these structures are provided on the table portion 101 , and the other parts are provided from below. It penetrates the table part 101 upward. At least the outer surface of each structure and the table portion 101 is formed of a material resistant to chemicals used for decontamination (for example, ethanol, hydrogen peroxide, sodium hypochlorite solution). In this embodiment, it is made of stainless steel alloy. The basic structure of the exterior structure part 10 is the same as that of a commercially available safety cabinet (therefore, regarding the exterior structure part 10, the description of the part not directly related to the present invention is omitted). The outer structure part 10 of the present embodiment has the shutter part 102 opened and closed by moving up and down in the lower area of the front part, but even if the shutter part 102 is opened (specifically, even if the lower area of the front part of the outer structure part 10 In the open state), the exterior structure part 10 of this embodiment is also configured so that the interior can be made into a clean environment. The degree of cleanliness is set to, for example, class A stipulated by PIC/S GMP. The shutter unit 102 is, for example, a flat member made of colorless and transparent tempered glass, and is opened and closed by, for example, an operator's hand. When the shutter portion 102 is opened, an opening is formed below the lower edge of the shutter portion 102 . Therefore, the lower area of the front part in the outer structure part 10 is opened. Materials (containers 51 to 54 and the like) can be carried in and out through the opening. In addition, a shutter opening sensor (not shown) is provided on the front portion of the exterior structure 10 , and when the shutter 102 is opened beyond a predetermined opening degree, an alarm is issued by displaying on the operation panel 21 or the like.

In front of the outer structure part 10 and behind (immediately behind) the gate part 102, a front suction part 103 opening upward is provided. The air part 103 sucks the airflow directed downward in the inside of the exterior structure part 10 (the space above the table part 101). The front air intake portion 103 is covered with a perforated plate or a metal mesh so as to allow ventilation in the vertical direction. The front air intake part 103 of this embodiment is strip-shaped, and is provided continuously on the left and right along the width direction of the front view of the shutter part 102. As shown in FIG.

When the operator loads materials into the exterior structure part 10, even if the shutter part 102 is opened to open the lower area of the front part of the exterior structure part 10 and form an opening, the airflow is sucked in through the front air suction part 103. , so that there is no air flow passing through the opening from the inside to the outside. Therefore, the cleanliness of the internal space of the exterior structure part 10 can be maintained.

Structures necessary for recovering cells are provided behind the front air suction part 103 of the outer structure part 10 (including a case where a part overlaps above the front air suction part 103).

The culture container stand 121 and the culture medium recovery container stand 123 are formed on the front panel 12 . The front panel 12 is formed of a vertically air-permeable material such as a perforated plate, and is installed so as to float above the table portion 101 . In this embodiment, the culture container 51, the cell recovery container 52, and the culture medium recovery container 54 are attached to the holding parts 61, 62, and 64 respectively on a single multi-well plate by mounting blocks for maintaining their positions. The culture container mount 121, the cell recovery container mount 122, and the culture medium recovery container mount 123 are formed on the same plane by dividing them by the shape stoppers. In one front panel 12, since a plurality of container mounts 121 to 123 are provided on the same plane, cleaning work, for example, is easy. A part of the front panel 12 is arranged on the table part 101 so as to cover a part of the front air intake part 103 from above. Since the front panel 12 is formed of a perforated plate, even in a positional relationship covering the front air intake portion 103 , it is difficult to hinder the intake of the airflow by the front air intake portion 103 . In addition, in this embodiment, the hole diameter of the perforated plate of the front panel 12 is formed larger than the hole diameter of the perforated plate of the front air intake part 103, and the ventilation property of an up-down direction is ensured. In addition, through such a positional relationship, there is an advantage that the space above the front air suction unit 103 can be effectively used as the culture container stand 121 and the culture medium recovery container stand 123 .

The manipulator 11 is provided inside the outer structure part 10 . The manipulator 11 is provided near the center of the front, rear, left, and right sides of the table portion 101 of the outer structure portion 10 . Therefore, structures necessary for cell recovery can be arranged in a balanced manner within the reach of the manipulator 11 , so that the cell recovery device 1 as a whole can be made more compact and can contribute to a reduction in occupied area.

As shown in Figures 4A to 4C and Figures 5 to 8 (in addition, Figures 4A to 4C and Figures 5 to 8 have some shape differences due to the relationship shown), the manipulator 11 has a front end capable of holding each Hands (or grips) 111 of the containers 51-54. The hand 111 has a pair of grip pieces 112 and 112 that approach and separate from each other. The movement at the time of approaching and separating is a linear movement in which the pair of grip pieces 112 and 112 maintain a parallel state. Each grip piece 112 is formed in a shape suitable for supporting a container. In each grip piece 112, a plurality of protrusions 113 to 113 are provided so as to protrude toward the inner side of the hand 111 in the approaching/separating direction. The hands 111 configured in this way not only hold the containers 51 to 54 (specifically, the holding parts 61 to 64 attached to the containers 51 to 54), but also hold the containers in a concave-convex fit state. Therefore, the manipulator 11 can reliably support the containers 51 to 54 even if the containers 51 to 54 are placed in various postures (for example, upside down, sideways) for stirring and discharging the contents.

The plurality of protrusions 113 to 113 provided on each grip piece 112 form a set of two, and each set is provided in two directions in an orthogonal relationship. Specifically, in the state shown in FIG. 4B , there are a group in which two convex portions are aligned in the vertical direction and a group in which two convex portions are aligned in the lateral direction. Thereby, each container 51-54 can be supported vertically and horizontally based on the hand 111. FIG. Moreover, when it is vertical, it can support upside down, and when it is horizontal, it can support upside down. Therefore, when transferring the liquid content to another container, the original container can be supported in the direction in which the liquid content is most easily transferred. Therefore, transfer work can be performed efficiently.

14A shows the first state, and FIG. 14B shows the second state of the state in which the manipulator 11 holds the culture container 51 to which the holder 61 is attached. The basic holding state is the first state. When transferring the contents to another container, when the mouth of the culture container 51 held by the manipulator 11 is tilted downward, and when the culture container 51 is shaken in the air , the manipulator 11 switches and grips the culture container 51 to be in the second state. The same applies to the containers 52 to 54 other than the culture container 51 . Switching between the first state and the second state is performed by, for example, temporarily placing the culture container 51 or the reagent container 53 on the temporary placing table 20 .

In addition, in order to facilitate recovery work when an abnormality occurs, the manipulator 11 is controlled to temporarily stop at the origin position (position where the hand 111 faces forward) during each operation.

In the cell recovery device 1 of the present embodiment, for example, commercially available containers 51 to 54 used for hand culture are used in a state where holding parts 61 to 64 are attached. By doing so, there is an advantage that it is not necessary to use a dedicated container of special shape. In each of the containers 51 to 54, a screw-type (rotary) cap is attached to a mouth formed to protrude in a cylindrical shape.

The holding parts 61 to 64 are members for enabling the robot hand 11 to easily hold the commercially available containers 51 to 54 . For example, the holding portion 61 for the culture container 51 is formed in the shape shown in FIGS. 10A to 10D , and becomes the shape shown in FIGS. 9A and 9B after being attached to the container. As for the holding parts 62-64 for other containers, the state which attached each container 52-54 or the state of a single body is shown in FIG. 11A-FIG. 13B.

Protruding portions 611 to 641 are provided on side surfaces of the respective holding portions 61 to 64 . The protrusions 611 to 641 are sandwiched between the pair of grip pieces 112 and 112 of the manipulator 11 . Further, recessed portions 612 to 642 corresponding to the convex portions 113 of the respective grip pieces 112 provided on the manipulator 11 are formed on the protruding portions 611 to 641 . If the manipulator 11 directly holds the respective containers 51 to 54 , the side portions of the respective containers 51 to 54 may bend and slip from the hand 111 of the manipulator 11 . On the other hand, by using the respective holding portions 61 to 64 as in this embodiment, the robot hand 11 can securely hold the respective containers 51 to 54 via the respective holding portions 61 to 64 . In addition, as shown in FIGS. 12A to 12C , the protruding portion 631 in the holding portion 63 for the reagent container 53 is not in the form of protruding from the main body of the holding portion 63 , but a recess is directly formed on the side of the main body of the holding portion 63 . 632.

As shown in FIG. 10C , a bottom recess 613 is formed at the bottom of the holding portion 61 for the culture container 51 . The same applies to the holding parts 62 to 64 for other containers, and bottom concave parts 623 to 643 are formed on the bottoms thereof. The bottom recesses 613 to 643 formed in the respective holding portions 61 to 64 are formed at asymmetrical positions with respect to axial symmetry (180 degrees) and rotational symmetry (90 degrees, 270 degrees). In this embodiment, a plurality of bottom recesses are formed, and the positions and/or sizes of the bottoms of the bottom recesses 613 to 643 are in different combinations depending on the types of containers 51 to 54 . Although the reagent container 53 is not shown in the figure, the positions of the respective bottom recesses 633 in the bottom are different for the cleaning solution (PBS) container 531 and the enzyme (trypsin) container 532 .

On the other hand, on each stand 121, 122, 123, 13, for example, a pin fixed on the front panel 12 is formed with a convex portion corresponding to the bottom concave portion 613-643 of each holding portion 61-64, and each container 51-64 If the 54 is accurately installed on each of the mounting platforms 121, 122, 123, and 13, the concave and convex will fit together. Through the combination of the unevenness, the respective containers 51 to 54 can be positioned at the precise positions of the respective mounting tables 121 , 122 , 123 , and 13 . In addition, it is possible to prevent misinstallation (the same container but in the wrong direction and when another container is installed).

In addition, in this embodiment, the cleaning solution (PBS) container 531 and the enzyme (trypsin) container 532 as the reagent container 53 have the same shape, and the holding part 63 for mounting the respective containers 531 and 532 has the same shape except for the bottom concave part 633. Also the same. However, since the bottom recesses 633 are formed at different positions, they can be distinguished from each other when they are attached to the reagent container stand 13 .

The fixing rings 71, 73 made of annular elastic body (silicone rubber, etc.) are hung on the culture container 51 (as shown in FIGS. and the reagent container 53 (as shown in FIGS. 12A to 12C ). Specifically, in the holding portion 61 for the culture container 51 , hook portions 614 , 614 are provided so as to protrude outward from a pair of opposing surfaces in the thickness direction. In the holding part 63 for the reagent container 53, the hook part 634 is provided so that it may protrude outward from one surface. Each hook part 614, 634 is formed in the shape whose diameter dimension of the front-end|tip expands. In the holding portion 61 for the culture container 51, as shown in FIGS. 9A and 9B , the fixing ring 71 is hung on a pair of hooks 614, 614 and the mouth of the culture container 51 in a ring shape or a figure-eight shape as shown in the drawing. (neck). In the holder 63 for the reagent container 53 , as shown in FIGS. 12A to 12C , the fixing ring 73 is hung on one hook 634 and the mouth (neck) of the reagent container 53 so as to go around the mouth. The culture container 51 and the reagent container 53 are held on the holding parts 61, 63 by the fixing rings 71, 73. Therefore, even if the manipulator 11 holds the holding parts 61, 63 and shakes them upside down, the containers 51, 53 will not move. The contents can be smoothly discharged from the respective containers 51 and 53 by dropping from the respective holding portions 61 and 63 using the manipulator 11 .

The culture container mounting table 121 is a table on which the culture container 51 for culturing cells with the holding unit 61 mounted thereon is placed. The culture container stand 121 is provided near the left and right center of the outer structure part 10 and between the shutter part 102 and the manipulator 11 . In the present embodiment, the culture container stand 121 is provided so as to substantially coincide with the center of the width of the shutter portion 102 .

As the culture container 51 of the present embodiment, commercially available "HYPERFlask™" (manufactured by Corning International Corporation) was used, and it was used with the holder 61 attached thereto as shown in FIGS. 9A and 9B . The culture container 51 has a substantially rectangular parallelepiped shape with a rectangular bottom, and its interior is divided into multiple layers in the thickness direction. The position of the mouth of the culture container 51 in the thickness direction is asymmetrical. A sensor for detecting the placement of the culture container 51 is provided in the section where each culture container 51 is placed in the culture container stand 121 . In this embodiment, the sensor uses an optical sensor that detects light by blocking it. The sensor is arranged at the bottom of the culture container platform 121, and the irradiation direction of the light is upward. In addition, the same sensors are provided on the reagent container set 13 , the cell recovery container set 122 , and the culture medium recovery container set 123 .

A plurality of (specifically, three) culture containers 51 can be vertically placed on the culture container stand 121 of the present embodiment. In addition, any number of culture containers 51 can be placed on the culture container stand 121 . In the present embodiment, any number of culture containers 51 within a range of one to three can be placed on the culture container stand 121 . Three culture containers 51 to 51 are arranged side by side in the left-right direction (linear direction) on the culture container stand 121 . That is, the culture container stand 121 is provided along the left-right direction (linear direction). Therefore, the three culture containers 51 to 51 are equidistant from the gate unit 102 . Therefore, there is no unevenness in the operator's ease of placement of the respective culture containers 51 on the culture container stand 121 , which is superior in operability, for example, compared to a configuration in which a part of the culture containers 51 have to be placed behind. In addition, since the culture container 51 can be quickly put into the cell recovery device 1 and quickly taken out, it is possible to avoid a decrease in cleanliness inside the cell recovery device 1 caused by taking out the culture container 51 . In the culture container setting table 121, the culture containers 51 are arranged so that the thickness direction runs front and rear, and the mouth portion which is asymmetrical in the thickness direction is located in the front.

For reasons such as incubator processing, the culture container 51 is temporarily taken out of the device by the operator during the processing (after the first step is completed), and is put back into the device after the processing is completed (before the second step is started). For the convenience of taking out and putting in, the culture container stand 121 is installed in the approximate center of the cell recovery device 1 and at the forefront. Such a positional relationship makes it possible to easily take out and put in the culture container 51 without polluting the inside of the cell recovery device 1 .

The cell recovery container mounting stand 122 is a stand on which the cell recovery container 52 which collects the cells taken out from the culture container 51 is mounted with the holder 62 attached thereto. In the cell recovery container 52, in addition to the cells, enzymes (trypsin) and a cleaning solution (PBS) for cleaning the inside of the culture container 51 are transferred to the inside of the culture container 51 in order to remove the cells from the cultured cells. The bonded surface (cell-attached surface) strips the enzymes added to the cells. The cell recovery container stand 122 is installed at the right rear of the culture container stand 121 (direction in plan view of the cell recovery device 1, hereinafter the same) and right front of the manipulator 11 (the reference of the rotation center of the manipulator 11 in plan view, the same hereinafter). The cell recovery container set 122 is provided on one front panel 12 together with the culture container set 121 and the culture medium recovery container set 123 .

The culture medium recovery container mounting stand 123 is a stand on which the culture medium recovery container 54 in which the culture medium taken out from the culture container 51 is recovered is mounted with the holder 64 attached thereto. The culture medium collection container stand 123 is provided at the right rear of the culture container stand 121 (between the culture container stand 121 and the cell recovery container stand 122 ) and in the right front of the manipulator 11 . The culture medium recovery container stand 123 is provided on one front panel 12 together with the culture vessel stand 121 and the cell recovery container stand 122 .

The reagent container mounting table 13 is a table on which the reagent container 53 in which the reagent for taking out the cells from the culture container 51 is placed is mounted with the holder 63 attached thereto. In addition, the reagent container 53 (in particular, the cleaning solution container 531 ) is also used to take out and recover the reagent (specifically, cleaning solution (PBS)) once placed in the culture container 51 . The reagent container stand 13 is provided at the left rear of the culture container stand 121 and at the left front of the manipulator 11 .

The reagent container set table 13 of the present embodiment is a two-layer type, the upper part 131 is a platform for placing the cleaning solution container 531 containing the cleaning solution (in this embodiment, PBS), and the lower part 132 is for placing the enzyme (PBS). In this embodiment, it is a stand on which the enzyme container 532 of trypsin is placed. The upper portion 131 and the lower portion 132 are supported by support shafts extending in the vertical direction. The upper layer part 131 and the lower layer part 132 can individually rotate around the support shaft 133 . The rotation range is the range indicated by the arrows in the illustration between the states shown by the solid line and the dashed-two dotted line in FIG. 2 . The state indicated by the solid line is the state at the rear position when the manipulator 11 holds the reagent container 53 . The state indicated by the dashed-two dotted line is the state at the front position when the operator carries in the reagent container 53 and places the reagent container 53 on the upper section 131 or the lower section 132 . By rotating, the upper part 131 and the lower part 132 of the reagent container stand 13 can be moved to the front position, so that the operator can easily place the reagent container 53 and the manipulator 11 can easily hold the reagent container 53 .

A flat rear panel 134 extending in the vertical direction is provided on the reagent container stand 13 (the upper section 131 and the lower section 132 ). A notched portion 135 is formed on the upper edge of the rear panel 134 so as to extend downward. The hook portion 634 having an enlarged diameter at the tip of the holding portion 63 attached to the reagent container 53 is sandwiched by the notch portion 135 . Thereby, the reagent container 53 is supported so that it will not fall even if the upper part 131 or the lower part 132 rotates.

In addition, in this embodiment, among the reagent containers 53, the number of cleaning solution containers 531 and the number of enzyme containers 532 are set to be equal to the number of culture containers 51 (arbitrary number, 1 to 3 in this embodiment). ) consistent. Therefore, since the number of reagent containers 53 required for cell recovery is unified, the operator can easily grasp the number of reagent containers 53 , and it is possible to prevent errors in the number of reagent containers 53 placed on the reagent container table 13 .

The lid opening and closing device 14 opens and closes the lids of the respective containers 51 to 54 . The cover opening and closing device 14 is provided at the left rear of the manipulator 11 . The lid opening and closing device 14 has a rotating grip portion 141 that rotates around a central axis extending in the vertical direction while holding the lid from the radially outward direction, thereby moving the lid relative to the containers 51-51. 54 body rotation. The rotating handle 141 is combined with a plurality of expanding pieces whose lower side expands or narrows, and moves up and down relative to the respective containers 51 to 54 placed on the container stand 142 provided at the lower part of the lid opening and closing device 14 . By rotating the grip portion 141 , a commercially available container with a screw cap can be used, so it is not necessary to prepare a special container having a cap with a special structure that is easy to open and close. As shown in FIG. 15 , the rotating grip part 141 is composed of upper and lower layers, the upper grip part 1411 has a smaller inner diameter, and the lower grip part 1412 has a larger inner diameter. Two types of caps having different outer diameters are used for the respective containers 51 to 54 used in the present embodiment. The cap C1 with a relatively small outer diameter is used for the culture container 51 and the reagent container 53 , and the cap C2 with a relatively large outer diameter is used for the cell recovery container 52 and the medium recovery container 54 . Therefore, the cap C1 having a relatively small outer diameter is held by the upper grip portion 1411 of the rotating grip portion 141 , and the cap C2 having a relatively large outer diameter is held by the lower grip portion 1412 . A container holding device 1421 is provided on the container setting table 142 . Some containers are held in the horizontal direction by the container holding device 1421 to open and close the lid by rotating the grip part 141 . Thereby, it is possible to stably open and close the lid.

The lid sensor 15 is a sensor for detecting the presence or absence of a lid on each of the containers 51 to 54, and in the present embodiment, an optical sensor whose irradiation direction of light is set downward is used. A cover opening and closing device 14 is provided at the right rear of the manipulator 11 . The respective containers 51 to 54 must be moved to the position of the lid sensor 15 after the opening and closing operation is performed by the lid opening and closing device 14 . Thereby, presence or absence of a cap of each container 51-54 can be detected reliably, and the actual state of opening and closing of each container 51-54 can be confirmed.

The cover stand 16 is a stand for temporarily placing the covers removed from the respective containers 51 to 54 . The capping stand 16 is provided at the rear right of the manipulator 11 and in front of the cap sensor 15 . The cover stand 16 has a fixed stand 161 immovable above the table portion 101 , and a movable stand 162 that can be attached to and detached from the fixed stand 161 . The caps removed from the respective containers 51 to 54 are placed on the moving table 162 . The moving table 162 is provided with the same portion as the protruding portions 611 to 641 of the respective holding portions 61 to 64, and this portion can be moved between the cover mounting table 16 and the cover opening and closing device 14 by being held by the hand 111 of the manipulator 11. .

The tilt table 17 can have the upper surface 171 in a horizontal state and a state in which the upper surface 171 is inclined obliquely forward. Fig. 1 shows a tilted state. The tilt table 17 is located on the right side of the manipulator 11 . By tilting the containers placed on the upper surface 171 of the tilt table 17 (specifically, the culture container 51, the reagent container 53, and the culture medium recovery container 54), it is possible to keep the liquid contents of other containers held by the manipulator 11 from It is effectively transferred to the respective containers 51 , 53 , 54 by spilling them.

The vibrating table 18 is a table that reciprocates (swings) left and right with respect to the table portion 101 . The vibrating table 18 is arranged at the right front of the manipulator 11, behind and to the right of the cell collection container setting table 122. The culture container 51 is placed horizontally on the vibrating table 18 so that the thickness direction thereof is vertical. By reciprocating the vibrating table 18 left and right while the culture container 51 is placed, impact can be applied to the culture container 51 . By this impact, in the inside of the culture vessel 51 in which either one or both of the cleaning solution (PBS) and the enzyme (trypsin) are placed, the cells can be separated from the surface (cells) to which the cultured cells are attached. Adhesive surface) peeled off. In the present embodiment, in the second step described later, the culture container 51 containing the cleaning solution is vibrated on the vibration table 18 .

The pipette device 19 is used to suspend the cell-containing content of the cell recovery container 52 . The pipette device 19 is arranged on the right side of the vibrating table 18 . The pipette device 19 mainly includes a pipette unit 191 , a connecting tube 192 , a hydraulic cylinder unit 193 , and a drip tray 194 . The pipette unit 191 has a cylindrical shape (specifically, a cylindrical shape) extending in the vertical direction and has a shape in which the lower end is narrowed. The pipette unit 191 is rotated and moved up and down by the swivel arm 195 supporting the upper end thereof, and rotates and moves up and down with respect to the rotation center in plan view. In addition, the pipette unit 191 is replaced every time a cell recovery operation is performed. The connection tube 192 connects the upper end of the pipette unit 191 and the hydraulic cylinder unit 193 in a breathable manner. The hydraulic cylinder part 193 is configured as a hydraulic cylinder extending in the vertical direction and reciprocating inside, and as the hydraulic cylinder part 193 reciprocates, the liquid is sucked into the internal space of the pipette part 191 through the lower end of the pipette part 191. , or discharge the liquid from the inner space of the pipette part 191 . The drip tray 194 is provided at a portion not overlapping the front panel 12 (the right side of the front panel 12 ) within the rotation range of the pipette unit 191 , and receives leaked liquid dripped from the pipette unit 191 .

The temporary placing table 20 is mainly a table with a flat upper surface on which the containers 51 to 54 are temporarily placed when the manipulator 11 alternately holds the containers 51 to 54 . In this embodiment, the temporary placement table 20 is installed at a position higher than the table part 101 .

The operation panel 21 is a touch panel, and displays the real-time operation status, and the operator can set the contents of each operation. In addition, an alarm display is performed when an abnormality occurs. In addition to this, the cell collection device 1 of the present embodiment is provided with a wired controller 22 for operating the manipulator 11 during maintenance and the like.

The various structures (including the respective sets 121, 122, 123, and 13) provided inside the outer structure part 10 used in the cell recovery work are installed so that they are within the movement range of the manipulator 11 (where the manipulator 11 can reach) Each container 51-54 can be grasped within the range), and each container 51-54 can be handled. The movement range of the manipulator 11 is an arc-shaped range. In particular, the positional relationship of each mounting table is as shown in FIG. 2 , and each mounting table is arranged at a position approximately the same distance from the rotation center of the manipulator 11 . Therefore, the work using the manipulator 11 is easier than when the objects to be grasped are located at scattered positions.

In addition, in particular, the reagent container stand 13 and the cell recovery container stand 122 are provided on the left and right sides of the culture container stand 121 (the sides of either the left or right) and the rear. Specifically, the reagent container stand 13 is provided on the left side of the culture container stand 121, and the cell collection container stand 122 is provided on the right side. Here, the cell recovery container 52, the reagent container 53, and the medium recovery container 54 are carried in before the cell recovery operation starts, and are carried out after the cell recovery operation is completed. Therefore, based on the relationship with the culture container 51 (preferably located at the frontmost position) that may be loaded and unloaded during the cell collection operation, the reagent container mounting table 13 and the cell collection container mounting table 122 are provided at a position away from the center of the cell collection apparatus 1. Moreover, it is set at the rear of the culture vessel setting table 121.

The placing stands 121 to 123 and 13 on which an operator places the respective containers 51 to 54 are provided at positions close to the table portion 101 in the height direction. Therefore, even if the operator is short, he can easily reach out, and does not hinder the work of placing the containers 51 to 54 on the respective stands 121 to 123 , 13 .

Next, an example of a series of operations of the cell recovery operation (first step, second step) performed by the cell recovery device 1 of this embodiment will be described. In addition, in the first step, culture containers 51 (three), reagent containers 53 (three wash solution (PBS) containers 531, three enzyme (trypsin) containers 532, ), the culture medium recovery container 54 (one) is carried into the device, and the situation after the state after being placed on each platform 121, 123, 13 will be described. In addition, in the second step, the culture containers 51 (three), the reagent containers 53 (three cleaning solution (PBS) containers 531 ), and the cell collection container 52 (one) that have passed the first step are loaded into the apparatus, and The cases after the state in which these are placed on the respective mounting stands 121 , 122 , 13 will be described. The following description of each step is performed in order of time.

In addition, all the movement of the containers 51 to 54 and the caps in the cell recovery apparatus 1 is performed by the robot arm 11 . In addition, opening and closing of the lids of the respective containers 51 to 54 is performed by moving the respective containers 51 to 54 relative to the lid opening and closing device 14 . The movement of the lid between the lid opening and closing device 14 and the lid placement stage 16 is performed by moving the moving stage 162 in the lid placement stage 16 by a robot. In addition, after the caps are opened and closed by the cap opening and closing device 14 , the openings of the containers 51 to 54 are shielded by the manipulator 11 below the cap sensor 15 to detect the state of opening and closing of the caps. In addition, although omitted in the description of the operation below, each container 51 to 54 is temporarily placed on the temporary placement table 20 when the robot arm 11 alternately holds the respective containers 51 to 54 .

First, the first step will be described. First, remove the cover of the culture medium recovery container 54 . The medium recovery container 54 with the cover removed is placed on the medium recovery container stand 123 (restored to the position before the movement). The removed cap is moved to the capping stage 16 . Next, the cap of the cleaning solution (PBS) container 531 is removed, and it is placed at a predetermined position on the upper section 131 of the reagent container stand 13 (the position placed before the movement, the same applies hereinafter). The removed cap is moved to the capping stage 16 . Then, the lid of the enzyme (trypsin) container 532 is removed, and it is placed at a predetermined position on the lower part 132 of the reagent container table 13 . The removed cap is moved to the capping stage 16 . Then, the cover of the culture container 51 is removed, and the culture container 51 is placed on the temporary placement table 20 . The removed cap remains in the state held by the cap opening and closing device 14 .

Next, the culture medium recovery container 54 is moved onto the tilt table 17, and the tilt table 17 is tilted so that the mouth of the culture medium recovery container 54 faces obliquely forward. The manipulator 11 holds the culture container 51 and tilts the culture container 51 , thereby transferring about half of the culture medium in the culture container 51 to the culture medium recovery container 54 . The manipulator 11 alternately holds the culture container 51 , and transfers the remaining medium in the culture container 51 to the medium recovery container 54 . Then, the culture container 51 is placed on the temporary placement table 20 . Next, the tilt table 17 is returned to a horizontal state, and the culture medium collection container 54 is moved to the culture medium collection container mounting table 123 on this basis.

In addition, when transferring the content of each container 51 to 54 to another container, in this embodiment, the movement of the content itself is not detected, and the control unit ensures that the time for moving the content is a constant time. way to control,

Next, the culture container 51 is moved to the tilt table 17, and the tilt table 17 is tilted so that the mouth of the culture container 51 faces obliquely forward. The manipulator 11 holds the cleaning solution (PBS) container 531 and transfers the cleaning solution (PBS) to the culture container 51 . The cleaning solution container 531 is placed at a predetermined position on the upper section 131 of the reagent container setting table 13 . Then, the tilt table 17 is returned to a horizontal state, and the lid of the culture container 51 is closed. Next, the manipulator 11 holds the culture container 51 and shakes it in the air. Then, the lid of the culture container 51 is opened, and placed at a predetermined position on the culture container stand 121 . Then, the empty cleaning solution container 531 is placed on the tilt table 17 (but not tilted). Next, the cleaning solution in the culture container 51 is transferred to the cleaning solution container 531 . Through this operation, the cleaning solution transferred to the culture container 51 returns to the original cleaning solution container 531 . The culture container 51 is placed at a predetermined position on the culture container stand 121 . The cleaning solution container 531 is placed at a predetermined position on the upper section 131 of the reagent container stand 13 .

Next, the culture container 51 is moved to the tilt table 17, and the tilt table 17 is tilted so that the mouth of the culture container 51 faces obliquely forward. The robot arm 11 holds the enzyme (trypsin) container 532 and transfers the enzyme (trypsin) to the culture container 51 . The enzyme container 532 is placed at a predetermined position on the lower section 132 of the reagent container table 13 . Then, the tilt table 17 is returned to a horizontal state, and the lid of the culture container 51 is closed. Next, the manipulator 11 holds the culture container 51 and shakes it in the air. Then, the culture container 51 is placed at a predetermined position on the culture container stand 121 . Next, the operator opens the shutter 102 to take out the culture container 51 , and then closes the shutter 102 . The culture container 51 temporarily taken out from the cell recovery apparatus 1 is moved by an operator to an incubator installed separately for processing.

Next, the lids of the medium recovery container 54 , the cleaning solution container 531 , and the empty enzyme container 532 are closed in this order. When the lid is closed, the moving table 162 is moved together with the lid from the lid mounting table 16 to the rotary gripping portion 141 of the lid opening and closing device 14 , and the containers 51 to 54 are moved to the container mounting table 142 of the lid opening and closing device 14 . Then, the caps held by the rotary grip 141 are attached to the respective containers 51 to 54 . At this point, the first step is completed.

Next, the second step will be described. Before the start of the second process, the operator opens the gate 102, places the culture container 51 processed in the incubator on the predetermined position of the culture container stand 121 (the position placed in the first process), and then closes the gate. 102. In the second step, first, the cap of the cell recovery container 52 is removed. And the lid that has been taken off is moved toward the lid setting platform 16 . Next, the cap of the cleaning solution (PBS) container 531 is removed, and it is placed at a predetermined position on the upper section 131 of the reagent container table 13 . The removed cap is moved toward the capping stage 16 . Then, the culture container 51 is shaken in the air, and then placed on the vibrating table 18 to operate the vibrating table 18 (tapping). Then, the culture container 51 is again shaken in the air. Then, the lid of the culture container 51 is removed, and the contents are transferred to the cell recovery container 52 . The removed cap is held by the cap opening and closing device 14 .

Next, the culture container 51 is moved to the tilt table 17, and the tilt table 17 is tilted so that the mouth of the culture container 51 faces obliquely forward. Next, the manipulator 11 holds the cleaning solution (PBS) container 531 and transfers the cleaning solution (PBS) to the culture container 51 . The lid of the culture container 51 is closed. Next, after shaking the culture container 51 in the air, it is placed on the vibrating table 18 and the vibrating table 18 is operated (impacted). Next, the culture container 51 is again shaken in the air. Then, the lid of the culture container 51 is removed, and the contents are transferred to the cell recovery container 52 . The lid of the culture container 51 is closed and moved to the culture container stand 121 . Then, the pipette unit 191 of the pipette unit 19 is inserted into the contents of the cell recovery container 52, and the hydraulic cylinder unit 193 is operated to suck and discharge the contents, thereby dispersing (piping). Then, the tip of the pipette unit 191 is lifted from the contents of the cell recovery container 52 and the hydraulic cylinder unit 193 is operated (idle rotation (idle rotation)), thereby removing the residual matter in the pipette unit 191. The (residual liquid) drops into the cell recovery container 52 . Next, the lid of the cell recovery container 52 is closed, and the lid of the cleaning solution container 531 is closed. The series of actions of the cell recovery device 1 is now over.

After the operator opens the shutter unit 102 and takes out the cell recovery container 52 and other containers from the cell recovery device 1, the cell recovery operation from the culture container 51 is completed. In addition, in the case of collecting cells from a plurality of (specifically, three in this embodiment) culture containers 51, first, the lids of all the reagent containers 53 are opened, and then the same series of operations as described above are performed. The treatment of each culture container 51 is repeatedly performed in a manner. After the treatment of all the culture containers 51 is completed, the lids of all the other cell recovery containers 52 and reagent containers 53 are closed.

According to the cell recovery apparatus 1 of the present embodiment configured as above, the culture container mounting table 121 is installed near the left and right center of the outer structure part 10 and at a position between the gate part 102 and the manipulator 11. It is easier to place and take the container 51 with respect to the culture container set table 121. Since the reagent container set table 13 and the cell recovery container set table 122 are arranged on the left, right, and rear sides of the culture container set table 121, at least each of the set tables 121, 122 can be placed. , 13 are compactly arranged inside the outer structure part 10 . Therefore, the operator only needs to take and put in the respective containers 51 to 54, the operation for cell recovery is automated, and the cell recovery device 1 can be made compact.

In addition, in the cell recovery device 1 of the present embodiment, the reagent container mounting table 13 can move back and forth inside the outer structure part 10 . Therefore, the operator can easily place the reagent container 53 , and the robot hand 11 can easily hold the reagent container 53 .

One embodiment of the present invention has been described above, but the present invention is not limited to the above-mentioned embodiment, and various changes can be made without departing from the gist of the present invention even if it is an unspecified form.

The structure and operation of the above-mentioned embodiment are summarized as follows. The cell recovery device 1 of the above-mentioned embodiment has: an outer structure part 10, which has a shutter part 102 that can be opened and closed by moving up and down on the front part, and can make the inside a clean environment; a robot arm 11 is provided on the outer structure part Inside of 10; the culture container 121 is placed on the culture container 51 for culturing cells; the reagent container 131, 132 is placed on the reagent container 53, wherein the reagent container 53 is put into the reagent for taking out the cells from the culture container 51; The cell recovery container setting table 122 is used for placing the cell recovery container 52 for recovering the cells taken out from the culture container 51, and each setting table is set so that the manipulator 11 can hold each container within the moving range of the manipulator 11 Inside the outer structure part 10, the culture container stand 121 is provided near the left and right center of the outer structure part 10 and at a position between the gate part 102 and the manipulator 11, and the reagent container The stands 131 , 132 and the cell recovery container stand 122 are provided on the left, right, and rear sides of the culture container stand 121 .

Thus, by disposing the culture container stand 121 at a position close to the left and right center of the outer structure part 10 and between the gate part 102 and the manipulator 11, the operator can easily place the culture container 51 on or from the culture container stand 121. 121, by placing the reagent container sets 131, 132 and the cell recovery container set 122 on the left, right, and rear sides of the culture container set 121, each set can be compactly arranged inside the outer structure part 10.

In addition, the reagent container mounts 131 and 132 can also move back and forth inside the outer structure part 10 .

Thus, by moving the reagent container mounting tables 131 and 132 back and forth, the operator can easily place the reagent container 53 , and the manipulator 11 can easily hold the reagent container 53 .

In addition, at the front of the outer structure part 10 and behind the gate part 102, a front air suction part 103 opening upward is provided, and the culture container stand 121 is provided on a front panel with air permeability in the vertical direction. 12. A part of the front panel 12 may also cover a part of the front air intake part 103 from above.

Accordingly, even in the positional relationship where the front panel 12 covers the front air intake section 103 , it is difficult to hinder the airflow intake at the front air intake section 103 . Furthermore, the space above the front air suction unit 103 can be effectively used at least as the culture container stand 121 .

As described above, according to the above-described embodiment, the operator can easily place the culture container 51 on or take it out from the culture container stand 121 , and can arrange each stand compactly inside the exterior structure portion 10 . Therefore, it is possible to provide a compact cell recovery device 1 that only needs to take out and put in the container, and the operation for cell recovery is automated.

Explanation of reference signs

1: Cell recovery device, 10: External structure department, 11: Manipulator, 12: Front panel, 13: Reagent container mounting table, 14: Lid opening and closing device, 15: Lid sensor, 16: Lid mounting table, 17: Tilting table, 18 : Vibration table, 19: Pipette device, 20: Temporary storage stand, 21: Operation panel, 22: Wired controller, 51: Culture container, 52: Cell recovery container, 53: Reagent container, 54: Medium recovery container , 61: holding part for culture container, 62: holding part for cell recovery container, 63: holding part for reagent container, 64: holding part for medium recovery container, 71: fixing ring (culture container), 73: fixing ring ( Reagent container), 101: workbench, 102: gate, 103: front suction part, 111: hand, 112: grip piece, 113: convex part, 121: culture container set, 122: cell recovery container set , 123: culture medium recovery container stand, 131: upper part (reagent container stand), 132: lower part (reagent container stand), 133: support shaft, 134: back plate, 135: notch part, 141: rotating grip Part, 142: container stand, 161: fixed stand (cover stand), 162: mobile stand (cover stand), 171: upper surface of the inclined stand, 191: pipette part, 192: connecting tube, 193: hydraulic cylinder part , 194: drip tray, 195: swivel arm, 531: washing solution (PBS) container, 532: enzyme (trypsin) container, 613: bottom recess (culture container), 623: bottom recess (cell recovery container), 633: Bottom concave part (reagent container), 643: bottom concave part (medium recovery container), 614: hook part (culture container), 634: hook part (reagent container), 1411: upper grip part, 1412: lower grip part, 1421: Container holding device.

Claims (3)

1. A cell recovery device comprising:

an external structure part which is provided with a gate part which is opened and closed by moving up and down on the front part and can enable the inside to be a clean environment;

a manipulator provided inside the outer structure portion;

a culture container table on which a culture container for culturing cells is placed;

a reagent container placing table for placing a reagent container containing a reagent for taking out a cell from the culture container;

a cell collection container table on which a cell collection container for collecting cells taken out from the culture container is placed,

each of the tables is provided inside the exterior structure so that the robot can hold each of the containers within a movement range of the robot,

the culture container stand is provided at a position between the gate section and the robot near the center of the outer structure section,

the reagent container stand and the cell collection container stand are provided on the left and right sides and the rear of the culture container stand.

2. The cell recovery apparatus according to claim 1, wherein the reagent container table is movable back and forth inside the external structure section.

3. The cell recovery device according to claim 1 or 2, wherein a front suction unit having an upper opening is provided in front of the outer structure unit and behind the shutter unit,

the culture container stand is provided on a front panel having ventilation in the vertical direction,

a part of the front panel is provided so as to cover a part of the front intake unit from above.

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