JP7440732B2 - Culture vessel support device - Google Patents

Description

The present invention relates to a culture vessel support device that supports a culture vessel for culturing cells, and more particularly to a culture vessel support device that can horizontally move and tilt the supported culture vessel.

In recent years, automated devices have been developed for cell culture in regenerative medicine to perform operations efficiently and aseptically. In operations necessary for culturing cells, it is necessary to inject and aspirate various liquids such as cell suspensions and culture fluids into the culture container. In particular, when the culture container is a well plate with a plurality of wells formed therein, a device for moving a nozzle relative to the well plate is required in order to sequentially inject or aspirate liquid into each well.
Therefore, a device has been proposed in which the above-mentioned operations are automated using a robot (for example, Patent Document 1). In the apparatus of Patent Document 1, a nozzle is supported by a three-axis orthogonal coordinate robot to inject or suck liquid into each well of a well plate. Further, the well plate can be tilted while being supported horizontally on the pedestal.

Patent No. 3413418

By the way, in devices equipped with complex mechanical structures such as the above-mentioned three-axis orthogonal coordinate robot, the size of the device increases, and the robot operates above the culture container, which generates dust from sliding parts. There is a problem that foreign substances such as dirt and abrasion powder get mixed into the culture container. In addition, in order to maintain sterility, it is necessary to perform decontamination periodically or as needed, so the 3-axis orthogonal coordinate robot is required to be resistant to decontamination agents and capable of decontamination. Another problem is that the shape is complicated and decontamination takes a long time. Furthermore, the motions performed by the three-axis orthogonal coordinate robot replace the motions performed by humans, and there is a problem in that there is a limit to the improvement of processing power.

In view of the above-mentioned circumstances, the present invention according to claim 1 is a culture container support device that supports and horizontally moves a culture container,
An X-axis guide rod arranged horizontally, a Y-axis guide rod arranged horizontally orthogonal to the X-axis guide rod, and a Y-axis that moves the X-axis guide rod along the orthogonal horizontal Y-axis. a moving mechanism, an X-axis moving mechanism for moving the Y-axis guide rod along the orthogonal horizontal X-axis, and an X-axis moving mechanism provided at the intersection of the X-axis guide rod and the Y-axis guide rod, A passive member that moves while being guided by the Y-axis guide rod as the X-axis guide rod moves, and also moves while being guided by the X-axis guide rod as the Y-axis guide rod moves by the X-axis moving mechanism. and a support member that is provided on the passive member and supports the culture container, and a tilt drive mechanism that tilts the support member,
The operation of the Y-axis moving mechanism, the X-axis moving mechanism, and the tilting drive mechanism is controlled by a control device to tilt and horizontally move the culture container supported by the support member. It is something.

According to such a configuration, it is possible to provide a culture container support device for horizontally moving a culture container, which is small in size, has a simple configuration, is easy to decontaminate, and is optimal for use in a sterile environment.

FIG. 1 is a sectional view of a cell processing device equipped with a culture vessel support device according to the present invention. FIG. 3 is a plan view showing a moving mechanism section of the culture container support device according to the present invention. 3 is a sectional view taken along line III-III in FIG. 2. FIG. FIG. 3 is a partial cross-sectional view showing a container support part of the culture container support device according to the present invention. FIG. 2 is an explanatory diagram illustrating operations required for culturing using a cell processing device equipped with a culture vessel support device according to the present invention.

Hereinafter, the present invention will be described with reference to the illustrated embodiment. In FIG. 1, a cell processing device 1 includes a culture container support device 4 that supports a well plate 3 as a culture container in a sterile working chamber 2A provided in an isolator 2; It is configured to include a dispensing device 6 holding a nozzle 5 as a liquid processing device.
The internal space of the isolator 2 is divided into upper and lower parts by a horizontal base table 2B, with the upper side of the base table 2B serving as the sterile working room 2A, and the lower side serving as the machine room 2C. The machine room 2C houses the drive units for the culture vessel support device 4 and the dispensing device 6, as well as a control device 7 that controls the operation of the cell processing device 1. Further, an air purifying device 8 is disposed above the sterile working chamber 2A to supply clean air to the sterile working chamber 2A from above.

The culture vessel support device 4 is provided with a vessel support part 4A on the top that supports the well plate 3, and the vessel support part 4A is arranged in a horizontal X-axis direction, a horizontal Y-axis direction orthogonal thereto, and a composite direction thereof. It is configured to include a moving mechanism section 4B at the lower part.
Further, in this embodiment, the well plate 3 supported by the culture vessel support device 4 has a total of six wells 3A formed by arranging three wells 3A in two rows, as shown in FIG. Each well 3A is a concave portion having the same volume and having a circular opening of the same size, and has a flat bottom. In such a well plate 3, cells are accommodated in each well 3A and cultured.
In addition, the container support part 4A may support a plurality of well plates 3 in parallel, and furthermore, a petri dish or a flask other than the well plate 3 may be supported as a culture container, These different types of culture vessels may be supported in parallel. Moreover, a culture container and a container other than the culture container can be supported in parallel.

The dispensing device 6 includes an elevating mechanism 9 for elevating the nozzle 5, and the nozzle 5 is held by an elevating member 9A of the elevating mechanism 9 so as to face vertically downward. Further, in the machine room 2C below the base table 2B, a servo motor 9B for driving the lifting mechanism 9 is arranged, and an air supply/exhaust device 10 is installed, and is connected to the nozzle 5 via a tube 10A. has been done. In the dispensing device 6, the nozzle 5 is a pipette, and the exhaust action of the supply/exhaust device 10 draws gas from the nozzle 5 to suck the liquid, and the air supply action supplies gas to the nozzle 5 to discharge the liquid. It has become.
In the dispensing device 6 configured as described above, the nozzle 5 is lowered by operating the elevating mechanism 9 and inserted into the well 3A of the well plate 3 located below, and the nozzle 5 is inserted into the well 3A by operating the air supply/exhaust device 10. Pipetting, which stirs the liquid, can be performed by suctioning and holding the liquid contained in the nozzle 3A and by discharging the liquid held in the nozzle 5 and injecting it into the well 3A. Further, the liquid contained in another container (not shown) supported simultaneously by the container support section 4A is sucked into the nozzle 5 and held therein, and the container support section 4A is horizontally moved by the moving mechanism section 4B, and the well plate 3 It is possible to perform dispensing to sequentially inject into a plurality of wells 3A.

FIG. 2 is a plan view showing the moving mechanism section 4B of the culture container support device 4, and FIG. 3 is a sectional view taken along line III--III in FIG. 2.
The moving mechanism part 4B that horizontally moves the container support part 4A that supports the well plate 3 includes an The Y-axis guide rod 14 is composed of two guide rods 13, 13 that are perpendicular to each guide rod 11, 11 of the X-axis guide rod 12 and arranged parallel and horizontally at a predetermined interval; A Y-axis moving mechanism 15 moves the rod 12 along a horizontal Y-axis perpendicular to the longitudinal direction of the guide rods 11 and 11, and a Y-axis moving mechanism 15 moves the Y-axis guide rod 14 along a horizontal Y-axis perpendicular to the longitudinal direction of the guide rods 13 and 13. An X-axis moving mechanism 16 for moving along the X-axis is provided at the intersection where each guide rod 11, 11 of the X-axis guide rod 12 and each guide rod 13, 13 of the Y-axis guide rod 14 intersect The passive member 17 is configured to be horizontally moved by the Y-axis moving mechanism 15 and the X-axis moving mechanism 16, and the passive member 17 is provided with the container support section 4A. This constitutes the culture container support device 4.

The X-axis guide rod 12 has one end of each guide rod 11, 11 fixed to a plate-shaped fixing member 12A, and the other end fixed to a plate-shaped fixing member 12B, and these fixing members 12A, 12B are arranged oppositely, and the pair of guide rods 11, 11 are of equal length and are maintained horizontally in parallel and at the same height with a constant interval. Similarly, the Y-axis guide rod 14 is configured such that one end of each guide rod 13, 13 is fixed to a plate-shaped fixing member 14A, and the other end is fixed to a plate-shaped fixing member 14B. The members 14A, 14B are arranged oppositely, and the paired guide rods 13, 13 are of equal length and are maintained horizontally in parallel and at the same height at regular intervals.
In this embodiment, the distance between the guide rods 11, 11 and the distance between the guide rods 13, 13 are equal, and the length of the guide rods 11, 11 is longer.

The X-axis guide rod 12 and the Y-axis guide rod 14 are shaped like a rectangular frame whose four corners are supported by four pillars 18 erected on the base table 2B in the sterile working room 2A. It is supported by a support member 19 so as to be horizontally movable.
The frame-shaped support member 19 has a pair of sides perpendicularly facing each guide rod 11 of the X-axis guide rod 12 among the four sides of the frame, and an X-axis support side 19A that supports the X-axis guide rod 12; 19B, and a pair of sides of the Y-axis guide rod 14 that are perpendicularly opposed to each guide rod 13, 13 are Y-axis support sides 19C, 19D that support the Y-axis guide rod 14. In this embodiment, The Y-axis support sides 19C and 19D are longer than the X-axis support sides 19A and 19B.
The upper surfaces of these X-axis supporting sides 19A, 19B and Y-axis supporting sides 19C, 19D are formed as horizontal surfaces, and the Y-axis rails 20A, 20B are provided for guiding along the horizontal Y-axis. Furthermore, X-axis rails 20C and 20D are provided on the upper surfaces of the Y-axis support sides 19C and 19D to guide the Y-axis guide rod 14 along the horizontal X-axis perpendicular to the guide rods 13 and 13.

The fixing members 12A, 12B of the X-axis guide rod 12 and the fixing members 14A, 14B of the Y-axis guide rod 14 are located outside the frame of the frame-shaped support member 19, respectively. Slide members 21A and 21B guided by Y-axis rails 20A and 20B are provided on each opposing surface, and slide members 21C guided by X-axis rails 20C and 20D are provided on each opposing surface of fixed members 14A and 14B. , 21D are provided. As a result, the X-axis guide rod 12 is movable along the horizontal Y-axis, and the Y-axis guide rod 14 is movable along the horizontal X-axis.
The guide rods 11, 11 of the X-axis guide rod 12 are arranged in parallel at the same height by fixing members 12A, 12B. Further, the guide rods 13, 13 of the Y-axis guide rod 14 are similarly arranged in parallel at the same height by the fixing members 14A, 14B, but as shown in FIG. Each guide rod 13, 13 of the Y-axis guide rod 14 is perpendicular to each guide rod 11, 11 at a higher position than each guide rod 11, 11 of the above-mentioned X-axis guide rod 12, and the above-mentioned Y-axis guide rod The fixing member 12A, 12B and fixing members 14A and 14B are set.

A passive member 17 is provided between the guide rods 11, 11 and between the guide rods 13, 13, which are the intersection points of the X-axis guide rod 12 and the Y-axis guide rod 14.
The passive member 17 includes a body part 17A formed as a regular square prism with rounded corners, and a body part 17A whose upper surface is formed by extending the four sides equally with respect to the cross section of the body part 17A at the upper part of the body part 17A. It is composed of a square stage section 17B larger than the cross section of the stage section 17B. The body portion 17A is formed to fit between each guide rod 11, 11 and between each guide rod 13, 13 with almost no gaps, and the stage portion 17B can have a container support portion 4A attached to its upper surface, and has an overhang. The lower surface engages with each guide rod 13, 13 located above each guide rod 11, 11, and is placed on each guide rod 13, 13 so that the passive member 17 is supported. ing.
According to such a passive member 17, when the X-axis guide rod 12 moves along the Y-axis, the body portion 17A of the passive member 17 is pushed by one of the guide rods 11, and each of the Y-axis guide rods 14 Guided by the guide rods 13, 13, it is moved in the front-rear direction of the Y-axis, and when the Y-axis guide rod 14 moves along the X-axis, the body portion 17A is pushed by one of the guide rods 13, It is guided by the respective guide rods 11, 11 of the X-axis guide rod 12, and is moved in the front-rear direction of the X-axis. By moving the X-axis guide rod 12 and the Y-axis guide rod 14 at the same time, the passive member 17 can be moved in the direction in which these movements are combined, and the passive member 17 can be moved not only in a straight line but also in a curved trajectory. 17 can be moved.
In this embodiment, the guide rods 11 and 13 are made of stainless steel, and the passive member 17 is made of a resin material as an engineering plastic that has sliding properties, wear resistance, and chemical resistance. It moves smoothly, does not generate foreign matter such as dust or abrasion powder, and can be decontaminated.

As the Y-axis moving mechanism 15, a Y-axis rack 22 is provided at the lower end of one fixed member 12A of the X-axis guide rod 12 in parallel with the Y-axis rail 20A. , a pinion gear 23A rotated by a servo motor 23 provided in a machine room 2C below the base table 2B is engaged with the pinion gear 23A. Further, as the X-axis moving mechanism 16, an X-axis rack 24 is provided at the lower end of one of the fixing members 14A of the Y-axis guide rod 14 in parallel with the X-axis rail 20C. A pinion gear 25A rotated by a servo motor 25 provided in the machine room 2C below the base table 2B is engaged with the pinion gear 25A.
As a result, by rotating the servo motor 23 in the forward and reverse directions, the X-axis guide rod 12 can be moved forward and backward in the Y-axis direction along the horizontal Y-axis, and by rotating the servo motor 25 in the forward and reverse directions. , the Y-axis guide rod 14 can be moved back and forth in the X-axis direction along the horizontal X-axis, and the passive member 17 and the container support section 4A can be moved in the horizontal direction.
In addition, as shown in FIG. 3, the Y-axis rack 22 and the X-axis rack 24 are installed at different heights so that they do not interfere with each other. In this embodiment, the Y-axis rack 22 is the same as the X-axis rack. It is located at a lower position than 24. Therefore, the position of the Y-axis rack 22 relative to the slide member 21A on the X-axis guide rod 12 is lower than the position of the X-axis rack 24 relative to the slide member 21C on the Y-axis guide rod 14, so that the fixed member 12A and the fixed member The mounting position for 14A is set.

FIG. 4 shows the container support part 4A of the culture container support device 4, with (a) showing the horizontal state and (b) showing the inclined state.
The container support section 4A includes a support member 26 that supports the well plate 3, a base member 27 on which the support member 26 is tiltably movable, and a tilt drive mechanism 28 that tilts the support member 26. A positioning portion 26A is formed on the upper surface of the support member 26, into which the well plate 3 is fitted and positioned.
The support member 26 is provided with a rotation shaft portion 26B at one end of the lower surface, and a horizontal support shaft 26Ba protrudes from the rotation shaft portion 26B toward both sides. A shaft support 27A that rotatably supports both ends of the support shaft 26Ba is provided at one end, and by pivotally supporting the support shaft 26Ba on the upper surface of the base member 27, the support shaft 26 is provided so as to be rotatable up and down.
The support shaft 26Ba is arranged parallel to the horizontal Y-axis or the X-axis, and in this embodiment, as shown in FIG. They are arranged in parallel.

As the tilt drive mechanism 28, a grooved cam member 28A constituting an opposite cam is provided on the lower surface of the support member 26 near the tip opposite to the base end where the rotation shaft portion 26B is provided, A groove 28Aa extending in a direction perpendicular to the support shaft 26Ba is formed in the grooved cam member 28A so as to descend toward the distal end side of the support member 26. Further, on the upper surface of the base member 27, a direct-acting forward/backward drive device 28B such as an air cylinder or an electric actuator for driving the opposite cam is mounted, with the forward/backward rod 28Ba thereof being horizontally directed toward the distal end side of the support member 26. arranged to move forward. A cam follower 28C is provided at the tip of the reciprocating rod 28Ba, and the cam follower 28C is configured to reciprocate while being engaged with the groove 28Aa of the grooved cam member 28A.
With this configuration, the cam follower is moved as shown in FIG. 4(b) by extending the advancing/retracting rod 28Ba by the advancing/retracting drive device 28B from the horizontal state of the support member 26 and well plate 3 shown in FIG. 4(a). 28C advances toward the distal end side of the support member 26, and the grooved cam member 28A is pushed upward according to the shape of the groove 28Aa. Accordingly, the support member 26 is rotated upward using the support shaft 26Ba of the rotation shaft portion 26B as a fulcrum, thereby tilting the well plate 3 supported thereon.

An example of operations required for culturing cells using the well plate 3 as a culture vessel using the cell processing apparatus 1 having the culture vessel support device 4 configured as described above will be described with reference to FIG. 5.
FIG. 5 shows an operation for collecting cultured cells, and FIG. 5(a) shows an operation for peeling off cells C adhering to the bottom of the well 3A of the well plate 3.
That is, in FIG. 5(a), culture solution F is accommodated in well 3A, and cells C are stuck to the bottom. With the well plate 3 positioned by the positioning portion 26A of the support member 26, the nozzle 5 consisting of a pipette is lowered by the operation of the elevating mechanism 9 in the dispensing device 6 under the control of the control device 7, and the culture solution in the well 3A is lowered. While submerging the tip 5A in F, the nozzle 5 repeatedly sucks and discharges a predetermined amount of the culture solution F by operating the supply/exhaust device 10 to stir it. At this time, under the control of the control device 7, the well plate 3 is positioned by operating the Y-axis moving mechanism 15 and the X-axis moving mechanism 16 of the drive mechanism section 4B so that the nozzle 5 is located off the center of the well 3A. From this position, the Y-axis moving mechanism 15 and the X-axis moving mechanism 16 are operated to horizontally move the well plate 3 so that the nozzle 5 rotates relatively within the well 3A. As a result, the culture solution F is repeatedly sucked and discharged to the entire area of the cells C adhering to the bottom of the well 3A, and the detachment of the cells C is promoted.

FIGS. 5(b) and 5(c) show an operation for collecting detached cells C in the well 3A.
That is, most of the cells C are detached by the detachment operation shown in FIG. ing. Therefore, as shown in FIG. 5(b), the tilt drive mechanism 28 in the container support section 4A is operated under the control of the control device 7 to tilt the support member 26 to bring the well plate 3 into the tilted state. In this state, in the dispensing device 6, the nozzle 5 is lowered by operating the elevating mechanism 9 and inserted into the well 3A, and the culture solution F and cells C are sucked into the nozzle 5 by operating the supply/exhaust device 10. At this time, the control device 7 operates the Y-axis moving mechanism 15 and the X-axis moving mechanism 16 of the moving mechanism section 4B to control the base end side of the support member 26 that constitutes the container support section 4A, where the rotation shaft section 26B is provided. Move the well plate 3 so that the end of the well 3A, which is to be The nozzle 5 is caused to suck the culture solution F and the cells C in this state.
Further, in this embodiment, as shown in FIG. 5(c), under the control of the control device 7, the nozzle 5 is raised by operating the lifting mechanism 9 to such an extent that it does not come out of the inclined well 3A, and the well plate 3 is raised. The Y-axis moving mechanism 15 of the drive mechanism section 4B or The X-axis moving mechanism 16 is operated to move horizontally, and in this positional relationship, the culture fluid F and cells C that have already been sucked and held are discharged. Thereafter, the Y-axis movement mechanism 15 or the X-axis movement mechanism 16 is operated to reciprocate the well plate 3 so that the nozzle 5 relatively reciprocates between the lowest point and the highest point. Suction at the lowest point and ejection at the highest point are repeated several times, and through this operation, cells C still attached to the bottom of the well 3 are washed away and collected.
After repeating suction and discharge multiple times, the well plate 3 is moved horizontally and the nozzle 5 is lowered again so that the tip 5A of the nozzle 5 is located at the lowest point of the inclined well 3A. Culture solution F and cells C are aspirated to recover cells C along with culture solution F.
In addition, regarding horizontal movement of the inclined well plate 3 to position the lowest point and the highest point in the well 3A with respect to the position of the nozzle 5, the support shaft of the rotation shaft portion 26B of the support member 26 is used. In this embodiment, the support shaft 26Ba is disposed orthogonal to the Y-axis guide rod 14 and parallel to the horizontal X-axis. , the support member 26 supporting the well plate 3 in an inclined state is moved in the horizontal Y-axis direction perpendicular to the support shaft 26Ba.

In addition, the above-mentioned embodiment describes the cell processing apparatus 1 which is provided with the dispensing apparatus 6 as a liquid processing apparatus in the sterile operation chamber 2A, and uses a pipette as the nozzle 5 to perform suction and discharge as an operation example. We have described the case where cells C are detached and collected by repeating pipetting, but culture solution F is prepared by storing it in a container such as a bottle, and culture solution F is sucked from this container and held in nozzle 5. It is also possible to operate the moving mechanism section 4B under the control of the control device 7 to perform a dispensing operation to discharge and inject into the well plate 3 or other culture container located below the nozzle 5. In place of the dispensing device 6, there is a liquid feeding device equipped with a nozzle 5 that directly sends other liquids such as the culture solution F and washing solution from the container to the culture container, or a liquid delivery device that is equipped with a nozzle 5 that directly sends other liquids such as the culture solution F and washing solution from the well 3A of the well plate 3. The culture vessel support device 4 of the present invention can also be applied to a cell processing device 1 equipped with a device such as an aspirator having a nozzle 5 for suctioning and collecting such substances.
In the cell processing device 1 equipped with the culture container support device 4 according to the present invention, the nozzles 5 in these liquid processing devices are moved up and down by the lifting mechanism 9, and in the horizontal direction, the culture containers such as the well plate 3 are moved horizontally. This prevents foreign matter such as dust and abrasion powder from entering the culture container. In addition, since the movement of the nozzle 5 relative to the culture container is divided into the vertical movement of the nozzle 5 and the horizontal movement of the culture container, it is possible to operate the nozzle 5 in parallel, allowing efficient cell processing. I can do it.

Furthermore, although the X-axis guide rod 12 and the Y-axis guide rod 14 are composed of two guide rods 11, 11 and guide rods 13, 13 in this embodiment, the number of guide rods is two. It is not limited to a book, and may be one or three. For example, one X-axis guide rod 12 and one Y-axis guide rod 14 are connected to the passive member 17 by making them orthogonal to each other at different heights. By penetrating the X-axis guide rod 12 and the Y-axis guide rod 14, as in the present embodiment, as one guide rod moves, the passive member 17 is guided by the other guide rod. Can be moved horizontally.
In either case, the components for moving the container support section 4A in the XY direction, including the X-axis guide rod 12, the Y-axis guide rod 14, and the frame-shaped support member 19, are mostly composed of rod-shaped members, It is small, has a simple configuration, and is easy to decontaminate, making it ideal for use in the sterile work room 2A, which is a sterile environment.

1. Cell processing device 2A.. Sterile working room 3. Well plate (culture container) 4. Culture container support device 5. Nozzle 6. Dispensing device 7. Control device 12. X-axis guide rod 14. Y-axis guide rod 15. Y-axis movement mechanism 16.X-axis movement mechanism 17.Passive member 26.Supporting member 28.Tilt drive mechanism

Claims (3)

A culture container support device that supports and horizontally moves a culture container,
An X-axis guide rod arranged horizontally, a Y-axis guide rod arranged horizontally orthogonal to the X-axis guide rod, and a Y-axis that moves the X-axis guide rod along the orthogonal horizontal Y-axis. a moving mechanism, an X-axis moving mechanism for moving the Y-axis guide rod along the orthogonal horizontal X-axis, and an X-axis moving mechanism provided at the intersection of the X-axis guide rod and the Y-axis guide rod, A passive member that moves while being guided by the Y-axis guide rod as the X-axis guide rod moves, and also moves while being guided by the X-axis guide rod as the Y-axis guide rod moves by the X-axis moving mechanism. and a support member that is provided on the passive member and supports the culture container, and a tilt drive mechanism that tilts the support member,
The operation of the Y-axis moving mechanism, the X-axis moving mechanism, and the tilting drive mechanism is controlled by a control device to tilt and horizontally move the culture container supported by the support member. Culture vessel support device. The X-axis guide rod and the Y-axis guide rod are both composed of two parallel guide rods,
The passive member is characterized in that it is interposed between each of the two guide rods at an intersection point where each of the two guide rods intersects, and is supported by either one of the two guide rods. The culture vessel support device according to claim 1. The support member is tilted by the tilt drive mechanism about a support shaft parallel to the horizontal Y axis or the X axis,
The culture vessel support according to claim 1 or 2, wherein the control device horizontally moves the culture vessel supported by the support member in a tilted state in a direction perpendicular to the support shaft. Device.

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