WO2021177048A1 - Dispositif de récupération de cellules - Google Patents

Dispositif de récupération de cellules Download PDF

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Publication number
WO2021177048A1
WO2021177048A1 PCT/JP2021/006089 JP2021006089W WO2021177048A1 WO 2021177048 A1 WO2021177048 A1 WO 2021177048A1 JP 2021006089 W JP2021006089 W JP 2021006089W WO 2021177048 A1 WO2021177048 A1 WO 2021177048A1
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WO
WIPO (PCT)
Prior art keywords
container
stand
culture
robot arm
cell recovery
Prior art date
Application number
PCT/JP2021/006089
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English (en)
Japanese (ja)
Inventor
昇平 杉本
古野　哲生
Original Assignee
ロート製薬株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by ロート製薬株式会社 filed Critical ロート製薬株式会社
Priority to CN202180017787.0A priority Critical patent/CN115279881A/zh
Publication of WO2021177048A1 publication Critical patent/WO2021177048A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • the present invention relates to a cell recovery device that automatically recovers cells.
  • the cells are seeded in a culture vessel such as a dome or a culture flask, and the cells are allowed to stand in an incubator to adhere to the culture vessel and cultured in a monolayer state. There is (monolayer culture).
  • a culture vessel such as a portrait or a culture flask
  • the cells are allowed to stand in an incubator to adhere to the culture vessel and cultured in a monolayer state.
  • the cell surface is washed with PBS or the like, the cells are peeled off from the culture vessel with an enzyme (for example, trypsin), and the required amount of cells is placed in a new culture vessel containing the medium. It is done by transferring.
  • an enzyme for example, trypsin
  • the present invention has an exterior portion having a shutter portion opened and closed by vertical movement on the front portion to create a clean environment inside, a robot arm provided inside the exterior portion, and a culture container for culturing cells.
  • a cell on which a culture container stand on which the cells are placed, a reagent container stand on which a reagent container for storing a reagent used for removing cells from the culture container is placed, and a cell recovery container for collecting cells taken out from the culture container are placed.
  • a collection container stand is provided, and each stand is provided inside the exterior portion so that the robot arm can grip each container within the moving range of the robot arm, and the culture container stand is the culture container stand.
  • the reagent container pedestal and the cell collection container pedestal are provided near the center of the left and right sides of the exterior structure and at a position between the shutter portion and the robot arm, and the reagent container pedestal and the cell collection container pedestal are laterally and rearward to the left and right of the culture container pedestal. It is a cell recovery device provided.
  • the reagent container stand may be movable back and forth inside the exterior portion.
  • a front intake portion opened upward is provided in front of the exterior portion and behind the shutter portion, and the culture container stand is provided on a front panel having air permeability in the vertical direction.
  • a part of the front panel may be provided so as to cover a part of the front intake portion from above.
  • FIG. 1 is a front view schematically showing a configuration of a cell recovery device according to an embodiment of the present invention.
  • FIG. 2 is a plan view showing the top portion of the exterior portion, which is not shown, in order to schematically show the internal configuration of the configuration of the cell recovery device.
  • FIG. 3 is a right side view showing the side portion of the external structure portion, which is not shown, in order to schematically show the internal configuration of the configuration of the cell recovery device.
  • FIG. 4A is a front view showing a hand portion (or grip portion) of the robot arm of the cell recovery device.
  • FIG. 4B is a right side view showing a hand portion (or grip portion) of the robot arm of the cell recovery device.
  • FIG. 4C is a view taken along the line cc of FIG.
  • FIG. 4B showing a hand portion (or grip portion) of the robot arm of the cell recovery device.
  • FIG. 5A is a front and right side perspective views showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 5B is a front view showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 6A is a rear view showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 6B is a plan view showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 7A is a bottom view showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 7B is a right side view showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 5A is a front and right side portion of the hand portion (or grip portion).
  • FIG. 5B is a front view showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 6A is a rear view showing one
  • FIG. 8A is a left side view showing one left and right side portion of the hand portion (or grip portion).
  • FIG. 8B is a perspective view of the back surface and the left side surface showing the left and right one side portions of the hand portion (or grip portion).
  • FIG. 9A is a front view showing the culture vessel used in the cell recovery device together with the holder.
  • FIG. 9B is a plan view showing the culture vessel used in the cell recovery device together with the holder in a state where the cap is removed.
  • FIG. 10A is a front view showing a holder for a culture container used in the cell recovery device.
  • FIG. 10B is a plan view showing a holder for a culture container used in the cell recovery device.
  • FIG. 10C is a bottom view showing a holder for a culture vessel used in the cell recovery device.
  • FIG. 10D is a left side view showing a holder for a culture vessel used in the cell recovery device.
  • FIG. 11A is a front view showing the cell collection container used in the cell collection device together with the holder.
  • FIG. 11B is a plan view of the holder alone.
  • FIG. 12A is a front view showing the reagent container used in the cell recovery device together with the holder.
  • FIG. 12B is a plan view showing the reagent container used in the cell recovery device together with the holder.
  • FIG. 12C is a rear view showing the reagent container used in the cell recovery device together with the holder.
  • FIG. 13A is a front view showing the culture medium recovery container used in the cell recovery device together with the holder.
  • FIG. 13B is a plan view of the holder alone.
  • FIG. 14A shows a state in which the robot arm of the cell recovery device holds a culture container to which a holder is attached, and shows the first state.
  • FIG. 14B shows a second state in which the robot arm of the cell recovery device holds a culture container to which a holder is attached.
  • FIG. 15 is a vertical cross-sectional view showing the relationship between the rotary grip portion and the cap (a solid line display is an example of a large cap and a two-dot chain line display is an example of a small cap) among the cap opening / closing devices of the cell collection device.
  • the cell recovery device 1 according to the embodiment of the present invention will be described.
  • the expression of the direction in the following corresponds to the direction shown in FIG. 1 with respect to "up and down”.
  • the lower side shown in FIG. 2 (the side with the shutter portion 102 shown in FIG. 3) is the front
  • the upper side (the side opposite to the shutter portion 102 shown in FIG. 3) is the rear.
  • “left and right” correspond to the directions shown in FIGS. 1 and 2.
  • the cell recovery device 1 of the present embodiment is a device only for recovering cultured cells. Therefore, unlike an apparatus that performs a series of cell cultures, it is basically unexpected to collect a large amount of cultured cells. 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 the present embodiment is configured as shown in FIGS. 1 to 3, and mainly includes an exterior portion 10, a robot arm 11, a culture container stand 121, a reagent container stand 13, and a cell recovery container stand. It includes 122. In addition to these, a medium recovery container stand 123, a cap opening / closing device 14, a cap sensor 15, a cap stand 16, a tilt stand 17, a shaking stand 18, a pipette device 19, a temporary stand 20, and an operation panel 21 are provided. In the cell recovery device 1, physically moving parts (robot arm 11, reagent container stand 13, cap opening / closing device 14, tilting table 17, shaking table 18, pipette device 19) are moved by a control unit (not shown). Be controlled.
  • a control unit not shown. Be controlled.
  • the exterior part 10 has a box shape and has a space inside, and various structures necessary for cell recovery are arranged in this space. A part of these structures is installed on the table part 101, and a part of the other part is installed on the table part 101, with respect to the table part 101 having an upwardly long rectangular flat surface inside the exterior part 10. Is installed so as to penetrate from the bottom to the top. At least the outer surface of each structure and the table portion 101 is formed of a material having chemical resistance to chemicals for decontamination (for example, ethanol, hydrogen peroxide, sodium hypochlorite aqueous solution). In this embodiment, it is made of stainless alloy.
  • the basic structure of the exterior part 10 is the same as the structure of a commercially available safety cabinet (for this reason, the description of the part not directly related to the present invention is omitted for the exterior part 10).
  • the exterior portion 10 of the present embodiment has a shutter portion 102 that is opened and closed by moving up and down with respect to a lower region of the front portion, but even if the shutter portion 102 is opened (specifically, the front portion of the exterior portion 10). Even if the lower area of the is open), it is configured so that the inside can be a clean environment.
  • the cleanliness is set to, for example, Grade A specified by PIC / S GMP.
  • the shutter portion 102 is, for example, a flat plate formed of colorless transparent tempered glass, and is opened and closed by, for example, a worker.
  • an opening is formed below the lower edge of the shutter portion 102. Therefore, the lower region of the front surface portion of the exterior portion 10 is opened. Materials (each container 51 to 54, etc.) can be carried in and out using this opening.
  • a shutter release sensor (not shown) is provided on the front portion of the exterior portion 10, and when the shutter portion 102 is opened beyond a predetermined opening degree, a warning is given by a display on the operation panel 21 or the like. It has become.
  • a front intake unit 103 opened upward is provided in front of the exterior unit 10 and behind (immediately after) the shutter unit 102, and the operator moves the shutter unit 102 upward.
  • the front intake portion 103 is covered with a perforated plate or wire mesh so as to be ventilated in the vertical direction.
  • the front intake portion 103 of the present embodiment has a band shape, and is provided continuously to the left and right along the width direction of the shutter portion 102 in the front view.
  • the structure required for cell recovery is provided behind the front inspiratory portion 103 in the external structure portion 10 (including the case where a part of the structure overlaps the front inspiratory portion 103).
  • the culture container stand 121 and the medium recovery container stand 123 are formed on the front panel 12.
  • the front panel 12 is made of a material having breathability in the vertical direction, for example, a perforated plate, and is provided so as to float above the table portion 101.
  • a block-shaped stopper that holds the position of each of the culture container 51, the cell recovery container 52, and the medium recovery container 54 while being attached to the holders 61, 62, 64 is attached to one perforated plate.
  • the culture container stand 121, the cell recovery container stand 122, and the culture medium recovery container stand 123 are formed on the same plane by being partitioned by. Since a plurality of container stands 121 to 123 are provided on the same plane in one front panel 12, for example, cleaning work is easy.
  • a part of the front panel 12 is arranged on the table portion 101 so as to cover a part of the front intake portion 103 from above. Since the front panel 12 is formed of a perforated plate, the suction of airflow by the front intake portion 103 is less likely to be hindered even in a positional relationship that covers the front intake portion 103.
  • the hole diameter of the perforated plate of the front panel 12 is formed to be larger than the hole diameter of the perforated plate of the front intake portion 103, so that air permeability in the vertical direction is ensured. Further, due to such a positional relationship, it is advantageous that the space above the front intake unit 103 can be effectively used as the culture container stand 121 and the medium recovery container stand 123.
  • the robot arm 11 is provided inside the exterior portion 10.
  • the robot arm 11 is provided near the center of the table portion 101 of the exterior portion 10 on the front, back, left, and right sides. Therefore, since the structures necessary for cell recovery can be arranged in a well-balanced manner within the reach of the robot arm 11, the cell recovery device 1 can be made compact as a whole and can contribute to reducing the occupied area.
  • the tip of the robot arm 11 is as shown in FIGS. 4A to 4C and 5 to 8 (note that there is a slight difference in shape between FIGS. 4A to C and FIGS. 5 to 8 due to the relationship shown in the drawings).
  • the hand portion (or grip portion) 111 capable of holding each of the containers 51 to 54 is provided in the above.
  • the hand portion 111 includes a pair of gripping pieces 112, 112 that approach and separate from each other.
  • the movement at the time of approaching / separating is a linear movement such that the pair of gripping pieces 112, 112 are kept in a parallel state.
  • Each grip piece 112 has a shape suitable for supporting the container.
  • Each grip piece 112 is provided with a plurality of convex portions 113 to 113 protruding inward in the hand portion 111 in the approaching / separating direction.
  • the hand portions 111 configured in this way not only sandwich the containers 51 to 54 (specifically, the holders 61 to 64 mounted on the containers 51 to 54), but also in a state of uneven fitting. Therefore, the robot arm 11 can reliably support the containers 51 to 54 even if the containers 51 to 54 are held in various postures (for example, upside down or sideways) for stirring and discharging the contents. ..
  • each gripping piece 112 The plurality of convex portions 113 to 113 provided on each gripping piece 112 are formed into a set of two, and each set is provided in two directions having an orthogonal relationship. Specifically, in the state shown in FIG. 4B, there are a set in which two are arranged in the vertical direction and a set in which the two are arranged in the horizontal direction. As a result, each container 51 to 54 can be supported vertically or horizontally with the hand portion 111 as a reference. Moreover, it can be supported in the upside-down direction in the vertical direction and in the left-right upside direction in the horizontal direction. Therefore, when transferring the liquid contents to another container, the original container can be supported in the direction in which the liquid contents are most easily transferred. Therefore, efficient transfer work is possible.
  • FIG. 14A is the first state
  • FIG. 14B is the second state.
  • the basic gripping state is the first state, but when the contents are transferred to another container, the culture container 51 held by the robot arm 11 is tilted until the mouth faces downward, and the culture container 51 is tilted.
  • the robot arm 11 changes the culture container 51 to the second state.
  • the switching between the first state and the second state is performed, for example, after the culture container 51 or the reagent container 53 is once placed on the temporary storage table 20.
  • the robot arm 11 is controlled so as to temporarily stop at the origin position (the position where the hand portion 111 faces forward) in each operation.
  • Holders 61 to 64 are for making it easier for the robot arm 11 to hold commercially available containers 51 to 54.
  • the holder 61 for the culture container 51 has the shape shown in FIGS. 10A to 10D, and when attached to the container, the holder 61 has the shape shown in FIGS. 9A and 9B.
  • the holders 62 to 64 for other containers the state of being attached to the respective containers 52 to 54 or the state of a single unit is shown in FIGS. 11A to 13B.
  • Protruding portions 611 to 641 are provided on the sides of the holders 61 to 64.
  • the protrusions 611 to 641 are sandwiched by a pair of gripping pieces 112 and 112 of the robot arm 11.
  • the protrusions 611 to 641 are formed with recesses 612 to 642 that coincide with the protrusions 113 provided on the gripping pieces 112 of the robot arm 11. If the robot arm 11 directly holds the containers 51 to 54, the side portions of the containers 51 to 54 may bend or slip off from the hand portion 111 of the robot arm 11. On the other hand, by using the holders 61 to 64 as in the present embodiment, the robot arm 11 can surely hold the containers 51 to 54 via the holders 61 to 64.
  • the protruding portion 631 of the holder 63 for the reagent container 53 is not in a protruding form in the main body of the holder 63 as shown in FIGS. 12A to 12C, and is directly directed to the side portion of the main body of the holder 63.
  • a recess 632 is formed.
  • a bottom recess 613 is formed at the bottom of the holder 61 for the culture container 51.
  • the holders 62 to 64 for other containers are also formed with bottom recesses 623 to 643 at the bottom.
  • the bottom recesses 613 to 643 formed in the holders 61 to 64 are formed at positions that are asymmetrical in line symmetry (180 degrees) and rotational symmetry (90 degrees, 270 degrees).
  • a plurality of bottom recesses are formed, and the positions and / or sizes of the bottom recesses 613 to 643 at the bottom are different combinations for each type of container 51 to 54.
  • the positions of the cleaning liquid (PBS) container 531 and the enzyme (trypsin) container 532 at the bottom of each bottom recess 633 are different.
  • each of the pedestals 121, 122, 123, 13 convex portions corresponding to the bottom recesses 613 to 643 of the holders 61 to 64 are formed by pins fixed to, for example, the front panel 12, and the pedestals 121 are formed.
  • the containers 51 to 54 are correctly attached to the, 122, 123, and 13
  • uneven fitting is performed.
  • the cleaning liquid (PBS) container 531 and the enzyme (trypsin) container 532 which are the reagent containers 53, have the same shape, and the holder 63 attached to each container 531 and 532 also excludes the bottom recess 633.
  • the outer shape is the same.
  • the bottom recess 633 is formed at a separate position, they can be distinguished from each other when mounted on the reagent container stand 13.
  • the fastening rings 71 and 73 made of a ring-shaped elastic body (silicon rubber or the like) are used by the operator. It has been handed over in advance (before being carried into the cell recovery device 1).
  • the holder 61 for the culture container 51 hook portions 614 and 614 are provided so as to project outward from a pair of surfaces facing each other in the thickness direction.
  • a hook portion 634 is provided so as to project outward from one surface.
  • Each hook portion 614, 634 has a shape in which the diameter dimension of the tip is enlarged.
  • the retaining ring 71 has a ring shape as shown on the figure or 8 on the pair of hook portions 614 and 614 and the mouth portion (neck portion) of the culture vessel 51. It is hung in the shape of.
  • the fastening ring 73 has a hook portion 634 and the mouth portion (neck portion) of the reagent container 53, and the mouth portion is fastened to the mouth portion (neck portion). It is handed over as if it were.
  • the culture container stand 121 is a stand on which the culture container 51 for culturing cells is placed with the holder 61 attached.
  • the culture container stand 121 is provided near the center of the left and right sides of the outer structure portion 10 and at a position between the shutter portion 102 and the robot arm 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.
  • the culture container 51 of the present embodiment a commercially available "Hyper Flask TM" (manufactured by Corning International Co., Ltd.) is used, and a holder 61 is attached and used as shown in FIGS. 9A and 9B.
  • the culture container 51 has a substantially rectangular parallelepiped shape with a rectangular bottom surface, and the inside is divided into multiple layers in the thickness direction. The position of the mouth of the culture vessel 51 in the thickness direction is asymmetric.
  • a sensor for detecting that the culture container 51 is placed is provided in a section of the culture container stand 121 where each culture container 51 is placed. In this embodiment, an optical sensor that detects by shielding light rays is used as this sensor. The sensor is provided at the bottom of the culture vessel stand 121, and the direction of irradiation of light rays is upward. Similar sensors are also provided in the reagent container stand 13, the cell recovery container stand 122, and the medium recovery container stand 123.
  • a plurality of (specifically, three) culture containers 51 can be vertically placed on the culture container stand 121 of the present embodiment.
  • An arbitrary number of culture containers 51 can be placed on the culture container stand 121.
  • an arbitrary number of culture containers 51 can be placed on the culture container stand 121 in the range of 1 to 3.
  • the three culture containers 51 to 51 are arranged side by side in the left-right direction (straight direction) on the culture container stand 121. That is, the culture container stand 121 is provided along the left-right direction (straight line direction). Therefore, the distances of the three culture containers 51 to 51 from the shutter portion 102 are equidistant.
  • the culture containers 51 are arranged on the culture container stand 121 so that the thickness direction is along the front and back and the mouth portion, which is asymmetric in the thickness direction, is located in the front.
  • the culture vessel 51 is temporarily taken out of the apparatus during the treatment (after the completion of the first step) by the operator for reasons such as incubator treatment, and then put into the apparatus again after the treatment is completed (before the start of the second step). Be done.
  • the culture container stand 121 is provided at the substantially central portion of the cell collection device 1 and at the frontmost portion. Due to such a positional relationship, the culture container 51 can be easily taken in and out without contaminating the inside of the cell recovery device 1.
  • the cell recovery container stand 122 is a stand on which the cell recovery container 52 for collecting cells taken out from the culture container 51 is placed with the holder 62 attached.
  • the cell collection container 52 contains an enzyme (tripsin) added to peel the cells from the surface (cell adhesion surface) to which the cells cultured inside the culture container 51 adhere, and the inside of the culture container 51.
  • the cleaning solution (PBS) to be washed is also transferred.
  • the cell collection container stand 122 is rear right of the culture container stand 121 (direction in the plan view of the cell recovery device 1; the same applies hereinafter) and front right of the robot arm 11 (reference of the center of rotation in the plan view of the robot arm 11; the same applies hereinafter). It is provided in.
  • the cell recovery container stand 122 is provided together with the culture container stand 121 and the medium recovery container stand 123 on one front panel 12.
  • the medium recovery container stand 123 is a stand on which the medium recovery container 54 for collecting the medium taken out from the culture container 51 is placed with the holder 64 attached.
  • the medium recovery container stand 123 is provided on the right rear side of the culture container stand 121 (between the culture container stand 121 and the cell recovery container stand 122) and on the right front of the robot arm 11.
  • the medium recovery container stand 123 is provided together with the culture container stand 121 and the cell recovery container stand 122 on one front panel 12.
  • the reagent container stand 13 is a stand on which the reagent container 53 containing the reagent used for extracting cells from the culture container 51 is placed with the holder 63 attached.
  • the reagent container 53 (particularly the cleaning liquid container 531) is also used for taking out and collecting the reagent (specifically, the cleaning liquid (PBS)) once put in the culture container 51.
  • the reagent container stand 13 is provided on the left rear side of the culture container stand 121 and on the left front side of the robot arm 11.
  • the reagent container stand 13 of the present embodiment is a two-stage type, the upper stage 131 is a table on which a cleaning liquid container 531 containing a cleaning liquid (PBS in this embodiment) is placed, and the lower portion 132 is an enzyme (in this embodiment). It is a table on which the enzyme container 532 containing trypsin) is placed.
  • the upper portion 131 and the lower portion 132 are supported by a support shaft extending in the vertical direction.
  • the upper portion 131 and the lower portion 132 can rotate around the support shaft 133 separately.
  • the rotation range is the range indicated by the illustrated arrow between the state shown by the solid line and the alternate long and short dash line in FIG.
  • the solid line display state is a state in which the robot arm 11 is in the rear position when holding the reagent container 53.
  • the state of the alternate long and short dash line display is a state in which the operator carries in the reagent container 53 and places the reagent container 53 in the upper portion 131 or the lower portion 132 in the front position. Since the upper portion 131 and the lower portion 132 of the reagent container stand 13 can be moved to the front positions by rotation, the operator can easily place the reagent container 53 and the robot arm 11 can easily hold the reagent container 53.
  • the reagent container stand 13 (upper stage 131 and lower stage 132) is provided with a flat plate-shaped back plate 134 extending in the vertical direction.
  • a notch 135 is formed on the upper end edge of the back plate 134 so as to extend downward.
  • a hook portion 634 having an enlarged tip diameter dimension of the holder 63 mounted on the reagent container 53 is sandwiched in the notch portion 135. As a result, the reagent container 53 is supported so that the upper portion 131 or the lower portion 132 does not fall even if it rotates.
  • the quantity of the cleaning liquid container 531 and the quantity of the enzyme container 532 are the quantity of the culture vessel 51 (arbitrary quantity, 1 to 3 in the present embodiment). It is set to match. Therefore, since the quantity of the reagent container 53 required for cell recovery is unified, it is easy for the operator to grasp the quantity of the reagent container 53, and it is possible to prevent the operator from making a mistake in the quantity of the reagent container 53 placed on the reagent container stand 13. ..
  • the cap opening / closing device 14 opens / closes the caps of each container 51 to 54.
  • the cap opening / closing device 14 is provided on the left rear side of the robot arm 11.
  • the cap opening / closing device 14 has a rotary grip portion 141 that rotates the cap with respect to the main body of each container 51 to 54 by rotating around a central axis extending in the vertical direction with the cap sandwiched from the outer diameter direction. ing.
  • the rotary grip portion 141 is a combination of a plurality of expansion pieces whose lower side expands or narrows, and is moved up and down with respect to each of the containers 51 to 54 placed on the container stand 142 provided at the lower part of the cap opening / closing device 14. Move.
  • the rotary grip portion 141 Since a commercially available container to which a screw cap is attached can be used by the rotary grip portion 141, it is not necessary to prepare a special container provided with a cap having a special structure for facilitating opening and closing.
  • the rotary grip portion 141 is composed of two upper and lower stages, the inner diameter of the upper grip portion 1411 is small, and the inner diameter of the lower grip portion 1412 is large.
  • Two types of caps having different outer diameters are used for each of the containers 51 to 54 used in the present embodiment.
  • the cap C1 having a relatively small outer diameter is used for the culture container 51 and the reagent container 53
  • the cap C2 having a relatively large outer diameter is used for the cell recovery container 52 and the medium recovery container 54.
  • the cap C1 having a relatively small outer diameter is sandwiched by the upper grip portion 1411 in the rotary grip portion 141, and the cap C2 having a relatively large outer diameter is sandwiched by the lower grip portion 1412.
  • the container stand 142 is provided with a container holding device 1421.
  • the cap of some containers is opened and closed by the rotary grip portion 141 while being horizontally sandwiched by the container holding device 1421. As a result, the cap can be opened and closed stably.
  • the cap sensor 15 is a sensor for detecting the presence or absence of a cap in each of the containers 51 to 54, and in the present embodiment, an optical sensor in which the irradiation direction of light rays is set downward is used.
  • the cap opening / closing device 14 is provided on the right rear side of the robot arm 11. After the opening / closing operation is performed by the cap opening / closing device 14, each container 51 to 54 is always moved to the position of the cap sensor 15. As a result, the presence or absence of the cap in each of the containers 51 to 54 is reliably detected, and it is confirmed that the containers 51 to 54 are actually opened and closed.
  • the cap stand 16 is a stand for temporarily placing the caps removed from the containers 51 to 54.
  • the cap stand 16 is provided on the right rear side of the robot arm 11 and in front of the cap sensor 15.
  • the cap stand 16 has a fixed base 161 that is immovable above the table portion 101, and a moving base 162 that is attached to and detached from the fixed base 161.
  • the cap removed from each of the containers 51 to 54 is placed on the moving table 162.
  • the moving table 162 is provided with a portion similar to the protruding portions 611 to 641 of the holders 61 to 64, and the portion is held by the hand portion 111 of the robot arm 11, so that the cap stand 16 and the cap opening / closing device 14 are provided. Move to and from.
  • the tilting table 17 can have the upper surface 171 in a horizontal state and a state in which the upper surface 171 is tilted so as to face diagonally forward.
  • FIG. 1 shows an inclined state.
  • the tilting table 17 is provided on the right side of the robot arm 11. By tilting each container (specifically, the culture container 51, the reagent container 53, and the medium recovery container 54) placed on the upper surface 171 of the tilting table 17, the robot arm 11 with respect to each of the containers 51, 53, 54. The liquid contents of other containers held in the medium can be efficiently transferred without spilling.
  • the shaking table 18 is a table that reciprocates (swings) to the left and right with respect to the table portion 101.
  • the shaking table 18 is provided on the right front side of the robot arm 11 and behind and to the right of the cell collection container stand 122.
  • the culture vessel 51 is placed on the shaking table 18 on its side so that the thickness direction is up and down.
  • the shaking table 18 reciprocates left and right with the culture vessel 51 mounted on the culture vessel 51, so that the culture vessel 51 can be impacted. By this impact, cells can be peeled off from the surface (cell adhesion surface) to which the cultured cells adhere inside the culture vessel 51 containing either the washing solution (PBS), the enzyme (trypsin), or both. can.
  • the culture vessel 51 containing the cleaning liquid is vibrated on the shaking table 18.
  • the pipette device 19 is used to suspend the contents containing cells in the cell collection container 52.
  • the pipette device 19 is provided on the right side of the shaking table 18.
  • the pipette device 19 mainly includes a pipette portion 191 and a connecting tube 192, a cylinder portion 193, and a drop tray 194.
  • the pipette portion 191 has a tubular shape (specifically, a cylindrical shape) extending in the vertical direction and a shape in which the lower end is narrowed.
  • the pipette portion 191 rotates and moves up and down in a plan view with respect to the center of rotation when the swivel arm 195 whose upper end is supported swivels and moves up and down.
  • the pipette portion 191 is replaced after each cell collection operation.
  • the connecting tube 192 connects the upper end of the pipette portion 191 and the cylinder portion 193 in a ventilated manner.
  • the cylinder portion 193 extends in the vertical direction and is provided with a cylinder that reciprocates inside. As the cylinder portion 193 reciprocates, the liquid is sucked into the internal space of the pipette portion 191 via the lower end of the pipette portion 191. Or, it is configured to discharge the liquid from the internal space of the pipette portion 191.
  • the drop tray 194 is provided in a portion of the rotation range of the pipette portion 191 that does not overlap with the front panel 12 (to the right of the front panel 12), and receives the drops dropped from the pipette portion 191.
  • the temporary stand 20 is mainly a stand having a flat upper surface on which the containers 51 to 54 are temporarily placed when the robot arm 11 changes the containers 51 to 54. In this embodiment, it is provided at a position higher than the table portion 101.
  • the operation panel 21 is a touch panel, and the real-time operating state is displayed, and the operator can set the content of each operation. In addition, a warning is displayed when an abnormality occurs.
  • the cell recovery device 1 of the present embodiment is provided with a wired controller 22 for operating the robot arm 11 at the time of maintenance or the like.
  • Each structure (including each stand 121, 122, 123, 13) provided inside the exterior portion 10 used in the cell recovery work is within the moving range of the robot arm 11 (the range within the reach of the robot arm 11).
  • Each container 51 to 54 can be gripped, and each container 51 to 54 is provided so as to be processable.
  • the moving range of the robot arm 11 is an arcuate range.
  • the positional relationship of each stand is as shown in FIG. 2, and they are arranged at substantially the same distance from the rotation center of the robot arm 11. Therefore, the work using the robot arm 11 is easier than when the gripping objects are in different positions.
  • the reagent container stand 13 and the cell recovery container stand 122 are provided on the left and right sides (either the left or right side) of the culture container stand 121 and at the rear.
  • the reagent container stand 13 is provided on the left side of the culture container stand 121
  • the cell recovery container stand 122 is provided on the right side.
  • the cell recovery container 52, the reagent container 53, and the medium recovery container 54 are carried in before the start of the cell recovery work, and are carried out after the completion of the cell recovery work. Therefore, in relation to the culture vessel 51 (preferably in the foremost position) where the cells are taken in and out during the cell recovery work, the culture vessel stand is located at a position away from the central portion of the cell recovery device 1. It is provided behind 121.
  • the stands 121 to 123, 13 on which the workers place the containers 51 to 54 are provided at positions close to the table portion 101 in the height direction. Therefore, even if the worker is a short person, it can be easily reached, and the work of placing the containers 51 to 54 on the stands 121 to 123, 13 is unlikely to be hindered.
  • a culture container 51 (3 pieces) containing cells after culturing, a reagent container 53 (3 wash solution (PBS) containers 531 and 3 enzyme (trypsin) containers 532), The following will be described after the medium recovery container 54 (1 piece) is carried into the apparatus and placed on the stands 121, 123, 13 respectively.
  • the culture container 51 (3 pieces), the reagent container 53 (3 wash solution (PBS) containers 531), and the cell recovery container 52 (1 piece) that have undergone the first step are carried into the apparatus.
  • the following will be described after the state in which the containers are placed on the stands 121, 122, and 13 is set. The following description in each step is described in chronological order.
  • the cap of the medium recovery container 54 is removed.
  • the medium recovery container 54 with the cap removed is placed on the medium recovery container stand 123 (returned to the position before moving).
  • the removed cap is moved to the cap stand 16.
  • the cap of the cleaning liquid (PBS) container 531 is removed, and the container is placed at a predetermined position (the position that was placed before the movement, the same applies hereinafter) of the upper portion 131 of the reagent container stand 13.
  • the removed cap is moved to the cap stand 16.
  • the cap of the enzyme (trypsin) container 532 is removed, and the container is placed at a predetermined position on the lower portion 132 of the reagent container stand 13.
  • the removed cap is moved to the cap stand 16.
  • the cap of the culture container 51 is removed, and the culture container 51 is placed on the temporary stand 20.
  • the removed cap is held by the cap opening / closing device 14.
  • the medium recovery container 54 is moved to the tilting table 17, and the tilting table 17 is tilted so that the mouth of the medium recovery container 54 faces diagonally forward.
  • the robot arm 11 holds the culture container 51, and by tilting the culture container 51, about half of the medium in the culture container 51 is transferred to the medium recovery container 54.
  • the robot arm 11 changes the culture container 51, the rest of the medium in the culture container 51 is transferred to the medium recovery container 54.
  • the culture container 51 is placed on the temporary stand 20.
  • the medium recovery container 54 is moved to the medium recovery container stand 123.
  • the culture container 51 is moved to the tilting table 17, and the tilting table 17 is tilted so that the mouth of the culture container 51 faces diagonally forward.
  • the robot arm 11 holds the cleaning liquid (PBS) container 531 and transfers the cleaning liquid (PBS) to the culture container 51.
  • the cleaning liquid container 531 is placed at a predetermined position on the upper portion 131 of the reagent container stand 13.
  • the cap of the culture vessel 51 is closed.
  • the robot arm 11 holds the culture container 51 and shakes it in the air.
  • the cap of the culture container 51 is opened and placed at a predetermined position on the culture container stand 121.
  • the empty cleaning liquid container 531 is placed on the tilting table 17 (but not tilted).
  • the cleaning liquid in the culture container 51 is transferred to the cleaning liquid container 531.
  • the cleaning liquid transferred to the culture container 51 is returned to the original cleaning liquid container 531.
  • the culture container 51 is placed at a predetermined position on the culture container stand 121.
  • the cleaning liquid container 531 is placed at a predetermined position on the upper portion 131 of the reagent container stand 13.
  • the culture container 51 is moved to the tilting table 17, and the tilting table 17 is tilted so that the mouth of the culture container 51 faces diagonally 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 portion 132 of the reagent container stand 13.
  • the cap of the culture vessel 51 is closed.
  • the robot arm 11 holds the culture container 51 and shakes it in the air.
  • the culture container 51 is placed at a predetermined position on the culture container stand 121.
  • the operator opens the shutter unit 102, takes out the culture container 51, and then closes the shutter unit 102.
  • the culture vessel 51 once taken out from the cell recovery device 1 is moved by an operator to a separately provided incubator for processing.
  • the cap is moved from the cap stand 16 together with the moving stand 162 to the rotary grip portion 141 in the cap opening / closing device 14, and each container 51 to 54 is moved to the container stand 142 in the cap opening / closing device 14. Then, the cap gripped by the rotary grip portion 141 is attached to each of the containers 51 to 54. This completes the first step.
  • the second step Before the start of the second step, the operator opens the shutter portion 102, places the culture container 51 treated by the incubator in a predetermined position (the position where it was placed in the first step) of the culture container stand 121, and then shutters. Part 102 is closed.
  • the cap of the cell collection container 52 is removed. The removed cap is moved to the cap stand 16.
  • the cap of the cleaning liquid (PBS) container 531 is removed, and the container is placed at a predetermined position on the upper portion 131 of the reagent container stand 13.
  • the removed cap is moved to the cap stand 16.
  • the culture vessel 51 is shaken in the air and then placed on the shaking table 18 to operate the shaking table 18 (tapping).
  • the culture vessel 51 is shaken again in the air.
  • the cap of the culture vessel 51 is removed, and the contents are transferred to the cell recovery vessel 52.
  • the removed cap is held by the cap opening / closing device 14.
  • the culture container 51 is moved to the tilting table 17, and the tilting table 17 is tilted so that the mouth of the culture container 51 faces diagonally forward.
  • the robot arm 11 holds the cleaning liquid (PBS) container 531 and transfers the cleaning liquid (PBS) to the culture container 51. Close the cap of the culture vessel 51.
  • the culture vessel 51 is shaken in the air and then placed on the shaking table 18 to operate the shaking table 18 (tapping). Then, the culture vessel 51 is shaken again in the air.
  • the cap of the culture vessel 51 is removed, and the contents are transferred to the cell recovery vessel 52.
  • the cap of the culture vessel 51 is closed, and the culture vessel 51 is moved to the culture vessel stand 121.
  • the pipette portion 191 of the pipette device 19 is inserted into the contents of the cell collection container 52, and the cylinder portion 193 is operated to suck and discharge the contents to disperse the contents (pipetting). Then, by pulling up the tip of the pipette portion 191 from the contents of the cell collection container 52 and operating the cylinder portion 193 (performing a blank shot), the residue inside the pipette portion 191 is put into the cell collection container 52. .. Next, the cap of the cell collection container 52 is closed, and the cap of the washing liquid container 531 is closed. This completes the series of operations in the cell recovery device 1.
  • the cell collection operation from the culture container 51 is completed.
  • the caps of all the reagent containers 53 are opened, and then the treatment of each culture container 51 is performed. , Repeat in the same manner as the above series of operations.
  • the caps of all the other cell recovery containers 52 and the reagent containers 53 are closed.
  • the culture container stand 121 is located near the center of the left and right sides of the outer structure portion 10 and between the shutter portion 102 and the robot arm 11. It is easy for the operator to place and take the culture container 51 with respect to the culture container stand 121, and the reagent container stand 13 and the cell recovery container stand 122 are on the left and right sides of the culture container stand 121. Since it is provided on the side and in the rear, at least the stands 121, 122, and 13 can be compactly arranged inside the exterior portion 10. Therefore, the operator only needs to put in and take out each of the containers 51 to 54, and the operation for collecting cells is automated, and the cell collecting device 1 can be made compact.
  • the reagent container stand 13 is movable back and forth inside the exterior portion 10. Therefore, the operator can easily place the reagent container 53, and the robot arm 11 can easily hold the reagent container 53.
  • an exterior portion 10 having a shutter portion 102 opened and closed by vertical movement on the front portion to create a clean environment inside, a robot arm 11 provided inside the exterior portion 10, and cells.
  • the culture container stand 121 on which the culture container 51 in which the cells were cultured is placed, the reagent container stands 131 and 132 on which the reagent container 53 containing the reagent used for taking out the cells from the culture container 51 is placed, and the culture container 51 are taken out.
  • the cell collection container stand 122 on which the cell recovery container 52 for collecting the collected cells is placed is provided, and each stand is placed inside the outer structure portion 10 and each container is placed in the robot arm 11 within the moving range of the robot arm 11.
  • the culture vessel stand 121 is provided so that the arm 11 can be gripped, and the culture container stand 121 is provided near the center of the left and right sides of the exterior portion 10 and at a position between the shutter portion 102 and the robot arm 11.
  • the reagent container stands 131 and 132 and the cell recovery container stand 122 are cell recovery devices 1 provided on the left and right sides and behind the culture container stand 121.
  • the culture container stand 121 is provided near the center of the left and right sides of the outer structure portion 10 and at a position between the shutter portion 102 and the robot arm 11, the work is performed on the culture container stand 121. It is easy for a person to place and remove the culture container 51, and the reagent container stands 131, 132 and the cell recovery container stand 122 are provided on the left and right sides and behind the culture container stand 121, so that each stand can be placed. , Can be compactly arranged inside the exterior portion 10.
  • the reagent container stands 131 and 132 may be movable back and forth inside the exterior portion 10.
  • the operator can easily place the reagent container 53, and the robot arm 11 can easily hold the reagent container 53.
  • a front intake portion 103 opened upward is provided in front of the exterior portion 10 and behind the shutter portion 102, and the culture container stand 121 is a front panel having air permeability in the vertical direction. 12 is provided, and a part of the front panel 12 may be provided so as to cover a part of the front intake portion 103 from above.
  • the suction of the air flow by the front intake unit 103 is unlikely to be hindered. Then, the space above the front intake unit 103 can be effectively used at least as the culture vessel stand 121.
  • each stand can be compactly arranged inside the outer structure portion 10. .. Therefore, it is only necessary for the operator to put in and take out the container, and the operation for collecting cells is automated, and the compact cell collecting device 1 can be provided.
  • Retaining ring (reagent container), 101 ... Table section, 102 ... Shutter part, 103 ... Front intake part, 111 ... Hand part, 112 ... Grip piece, 113 ... Convex part, 121 ... Culture container stand, 122 ... Cell recovery container stand, 123 ... Medium recovery container stand, 131 ... Upper part ( Reagent container stand), 132 ... Lower part (Reagent container stand), 133 ... Support shaft, 134 ... Back plate, 135 ... Notch, 141 ... Rotating grip, 142 ... Container stand, 161 ... Fixing stand (cap stand), 162 ... Moving table (cap stand), 171 ... Upper surface of tilting table, 191 ... Pipet part, 192 ...

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Abstract

Dispositif de récupération de cellules étant équipé des éléments suivants : une partie extérieure capable de créer un environnement propre à l'intérieur, la partie extérieure ayant une partie d'obturation qui s'ouvre et se ferme en se déplaçant vers le haut et vers le bas sur la partie avant ; un bras de robot ; un support de récipient de culture sur lequel est placé un récipient de culture dans lequel des cellules sont cultivées ; un support de récipient de réactif sur lequel est placé un récipient de réactif ; et un support de récipient de récupération de cellule sur lequel est placé un récipient de récupération de cellule pour récupérer des cellules retirées du récipient de culture. Chaque support est disposé à l'intérieur de la partie extérieure de telle sorte que chaque récipient peut être saisi à l'intérieur de la plage de déplacement du bras de robot. Le support de récipient de culture est disposé à proximité du centre des côtés droit et gauche de la partie extérieure et à un emplacement entre la partie d'obturateur et le bras de robot. Le support de récipient de réactif et le support de récipient de récupération de cellule sont disposés sur le côté et derrière le support de récipient de culture.
PCT/JP2021/006089 2020-03-04 2021-02-18 Dispositif de récupération de cellules WO2021177048A1 (fr)

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JP2020036640A JP7422564B2 (ja) 2020-03-04 2020-03-04 細胞回収装置
JP2020-036640 2020-03-04

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006115798A (ja) * 2004-10-25 2006-05-11 Kawasaki Heavy Ind Ltd オートクレーブ滅菌を利用した自動細胞培養装置及びその使用方法
WO2016104666A1 (fr) * 2014-12-26 2016-06-30 テルモ株式会社 Procédé de transport de liquide
WO2016170623A1 (fr) * 2015-04-22 2016-10-27 株式会社安川電機 Système de culture cellulaire et procédé de culture cellulaire
WO2017069148A1 (fr) * 2015-10-20 2017-04-27 ロート製薬株式会社 Dispositif de traitement de cellules
JP2018000020A (ja) * 2016-06-28 2018-01-11 株式会社デンソーウェーブ 細胞培養容器，自動細胞培養装置，液体収容器，ロボットハンド及びロボットシステム

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2012147685A (ja) 2011-01-17 2012-08-09 Tokyo Women's Medical College 細胞培養処理システム

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006115798A (ja) * 2004-10-25 2006-05-11 Kawasaki Heavy Ind Ltd オートクレーブ滅菌を利用した自動細胞培養装置及びその使用方法
WO2016104666A1 (fr) * 2014-12-26 2016-06-30 テルモ株式会社 Procédé de transport de liquide
WO2016170623A1 (fr) * 2015-04-22 2016-10-27 株式会社安川電機 Système de culture cellulaire et procédé de culture cellulaire
WO2017069148A1 (fr) * 2015-10-20 2017-04-27 ロート製薬株式会社 Dispositif de traitement de cellules
JP2018000020A (ja) * 2016-06-28 2018-01-11 株式会社デンソーウェーブ 細胞培養容器，自動細胞培養装置，液体収容器，ロボットハンド及びロボットシステム

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JP7422564B2 (ja) 2024-01-26
JP2021136901A (ja) 2021-09-16

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