JP5270967B2 - Automatic cell culture equipment - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an automatic cell culture apparatus that operates a plurality of culture vessels collaterally and carries out a part of cull operation without involvement of operation robot. <P>SOLUTION: The operation part 12 of the automatic cell culture apparatus is equipped with an operation station 20 having five stages 21-25, five dishes corresponding to each stage are placed on one dish table 26, which is installed on the operation station 20. The dish is operated by the operation stages 21, 22 and 25 for carrying out necessary operations for cell culture among the stages 21-25 and each dish is moved to an adjacent stage by the rotation of the dish table 26. A series of operations for the five dishes is completed by repeating the operations. <P>COPYRIGHT: (C)2010,JPO&amp;INPIT

Description

  The present invention relates to an automatic cell culture apparatus, and more specifically, a plurality of culture containers sequentially move on a plurality of stages, so that a series of operations required for each culture container can be performed in parallel. The present invention relates to an automatic cell culture apparatus.

  Conventionally, an automatic cell culture apparatus equipped with an operation robot has been put into practical use for automatically culturing cells, tissues, etc. of human skin, cartilage, bone, blood vessels, organs and the like (for example, Patent Document 1). ). With the development of an automatic cell culture device equipped with an operation robot, it is possible to provide regenerative medicine safely and at low cost by preventing contamination and cross-contamination of cultured cells.

However, in an automatic cell culture apparatus equipped with such an operation robot, the operation robot performs most of a series of operations such as medium exchange operation and subculture operation necessary for culture, and these operations require a long time. There's a problem. That is, operations such as taking out a culture container such as a dish, opening and closing the lid of the culture container, taking out a pipettor tip, attaching and removing a tip to and from the pipetter are performed individually for each culture container, Since the operation robot also performs removal and storage, opening and closing of the lid of the chemical solution container, etc., there may be a problem that the operations for all the culture containers cannot be completed within a predetermined time.
JP 2008-54690 A

  The present invention has been made to solve the above-described problems of the prior art, and an object of the present invention is to provide an automatic cell culture apparatus capable of sequentially operating a plurality of culture vessels in parallel. Is to provide.

The automatic cell culture device of the present invention is an automatic cell culture device including an operation unit that performs a series of operations necessary for cell culture on a plurality of culture vessels, wherein the operation unit includes the plurality of cultures. An operation stage on which a table for holding each container is placed and an operation station having a plurality of stages is provided, and at least a part of the plurality of stages performs at least one of the plurality of operations. The plurality of culture containers are moved to the next stage after the operation is performed, and the operations are performed in parallel in the plurality of stages, and the plurality of culture containers are moved to the next stage. The movement is performed by the table, the table holds the plurality of culture vessels arranged in a circle, and the table has a plurality of trays on which the culture vessels are placed, The plurality of culture containers are further held by the table via the tray and further include an operation robot for performing operations necessary for cell culture in the automatic cell culture apparatus, and the operation robot is connected to the operation station. The table is installed and the table is taken out from the operation station .

  In the above, the operation in the operation stage includes, for example, opening / closing operation of the lid of the culture vessel, tilting operation on the culture vessel, pipetting operation on the culture vessel, rotation operation on the culture vessel, and the culture vessel Swinging operation, tapping operation of the culture vessel, and microscopic observation of the cells in the culture vessel.

  The table of the present invention is for holding the plurality of culture vessels in each of the plurality of stages in an operation station, holding the plurality of culture vessels in the plurality of stages, and the next stage of the plurality of culture vessels Used in the automatic cell culture device described above.

  The table may include a plurality of trays on which culture containers are placed, and the plurality of culture containers may be held on the table via the trays.

  According to the automatic cell culture apparatus of the present invention, a plurality of culture containers sequentially move on a plurality of stages, so that a series of operations necessary for each culture container can be performed in parallel, so that all Compared to a conventional automatic cell culture apparatus that performs a culture operation for each culture vessel one by one, the time to complete the operation can be greatly shortened, and the operation efficiency can be greatly improved.

  Embodiments of the present invention will be described with reference to the drawings, but the present invention is not limited to the following embodiments.

  FIG. 1 is a plan view showing an entire automatic cell culture apparatus according to an embodiment of the present invention. The automatic cell culture apparatus according to the present embodiment includes a housing 10, an operating robot 11 installed in the housing 10, a horizontal axis 13 for moving the operating robot 11, and various operations necessary for cell culture. An operation unit 12 for performing the operation and an incubator 14 for performing cell culture in the culture vessel are provided. The operation robot 11 in this embodiment is an articulated robot provided with two fingers that hold a culture container, a drug container, or the like. A refrigerated storage (not shown) for storing various drugs at a low temperature is installed under the incubator 14, and a centrifuge (for separating cultured cells) is provided under the operation unit 12. (Not shown) is installed. A room temperature storage shelf 15 for storing various drugs at room temperature is provided on the left side of the inside of the housing 10, and the room temperature storage shelf 15 includes a culture instrument such as a dish or pipette tip, a drug, a medium, and the like. Are stored, and a warehousing mechanism (not shown) for performing these warehousing is provided. Further, on the right side of the inside of the housing 10, there are provided a cell loading / unloading unit 16 for loading and unloading cells, a trash box (not shown) for discarding the used culture apparatus, and the like. A compressor 17 for driving the automatic cell culture device, a nitrogen gas generator 18 used for sterilization work in the housing 10 and the like are provided on the left outer side of the housing 10.

  In the automatic cell culture device of this embodiment, the operation unit 12 temporarily places an operation station 20 for performing various operations on the culture vessel, a pipette device 54, a centrifuge tube, a drug container, and the like. And a capper 53 for opening and closing caps such as a centrifuge tube and a medicine container.

  A plan view of the operation station 20 in this embodiment is shown in FIG. The operation station 20 includes five stages from first to fifth stages 21 to 25, and a dish table 26 is placed on these stages 21 to 25. FIG. 3A shows a plan view of the dish table 26, and FIG. 3B is a cross-sectional view taken along the line BB in FIG. As shown in FIG. 3A, the dish table 26 is provided with five opening windows 27, 27... Corresponding to the above five stages 21 to 25. Each opening window 27 has a dish tray 57. It is placed. 4A is a plan view of the dish tray 57, and FIG. 4B is a side view of the dish tray 57. FIG.

  As shown in FIG. 4B, the dish tray 57 of the present embodiment has an opening 58 at the center, and an engaging portion 58 having an L-shaped cross section is provided around the opening 58. A dish for cell culture is held by the locking portion 59. The dish tray 57 is provided with two locking members 61, and the locking member 61 engages with a locking hole 62 shown in FIG. It is held on the table 26. Further, the dish tray 57 is provided with an attachment 60, and the operation robot 11 described above can carry the attachment 60 by gripping the attachment 60.

  Further, as shown in FIG. 3B, a recess 28 in which the thickness of the dish table 26 is reduced is provided on the outer periphery of the opening window 27 of the dish table 26. The locking part 59 is placed. Therefore, the dish is held on the stages 21 to 25 via the dish tray 57. Therefore, each culture container can be moved to the next stage by the rotation of the dish table 26. The dish table 26 is provided with a number of locking holes 62 that engage with the locking members 61 of the dish tray 57. Further, as shown in FIG. 3 (b), an attachment 29 is attached to the lower surface of the dish table 26, and the operating robot 11 holds the attachment 29, thereby operating the five dishes once. It is possible to carry them together.

  In the present embodiment, among the stages 21 to 25, the first stage 21, the second stage 22, and the fifth stage are operation stages that operate on the dish, and the third and fourth stages 23. And 24 are stages where no operation is performed on the dish.

  In the first stage 21 of the present embodiment, four operations are performed: a dish lid opening / closing operation, a dish tilting operation, a pipetting operation, and a dish rotating operation.

  FIG. 5 is a right side view of the operation station 20 of FIG. As shown in FIGS. 1 and 5, the first stage 21 includes a stepping motor 32 fixed on a block 30 and a turning arm 31 that can be turned by the stepping motor 32. A suction cup unit 33 is attached. The suction cup unit 33 is provided with a suction cup pad 34 for sucking and opening and closing the lid 50b of the dish 50, and further provided with a contact switch 35 and a detection plate 36. A vacuum is supplied to the suction pad 34 from a vacuum source. The suction of the dish lid 50b by the suction pad 34 is detected when the suction pad 34 has a negative pressure. That is, when the dish lid 50b is correctly positioned, the contact switch 35 and the detection plate 36 maintain a predetermined distance, but when the dish lid 50b rides on the dish body 50a and is not properly closed, the suction cup When the pad 34 and the detection plate 36 are raised and contact the contact switch 35, a failure in the operation of closing the lid is detected. In this embodiment, since the position of the suction pad 34 needs to be adjusted according to the thickness of the dish, the vertical position of the stepping motor 32 can be adjusted by changing the height of the block 30 using an air cylinder. It is configured.

  6A and 6B are cross-sectional views taken along line P-P in FIG. As shown in FIG. 5 and FIG. 6A, a stepping air cylinder 38 with a guide and a stepping unit attached to the first stage 21 of the present embodiment so as to be movable up and down by driving the air cylinder with guide 38 are provided. A motor 39 is provided, and a rotating plate 40 is attached to the stepping motor 39. In the present embodiment, after the dish 50 is lifted by raising the guided air cylinder 38, the dishing 50 is rotated in the horizontal plane by the rotation of the stepping motor 39.

  Further, as shown in FIG. 6A, the first stage 21 of the embodiment is provided with a ROBO cylinder 41, and the ROBO cylinder 41 has two atlases 42 (1 in FIG. 6B). Only the book is shown). The dish body 50 a is inclined by the upper and lower portions of the ROBO cylinder 41, and the contents can be easily sucked out by the chip 43. The degree of inclination of the dish body 50a can be adjusted by moving the robot cylinder 41 up and down as shown in FIG. 6 (b).

 FIG. 7 is a side view taken along the line QQ in FIG. 2 and shows the second stage 22 in the present embodiment. In the second stage 22, a swing operation is performed by tilting and rotating the dish 50. As shown in FIG. 7, the second stage 22 is provided with an air cylinder 44 with a guide and a stepping motor 45 attached so as to be movable up and down by driving the air cylinder 44 with a guide. A swing arm 46 and a ball plunger 47 are attached to the motor 45. In the present embodiment, after the ball plunger 47 tilts the dish 50 by the ascending air cylinder 44 with guide, the dish 50 is swung by the rotation of the stepping motor 45.

  In the operation station 20 of this embodiment, no particular operation is performed on the third stage 23 and the fourth stage 24, and the presence or absence of the dish 50 is detected by the fiber sensor 51 (FIG. 5). Only.

  FIG. 8 is a front view of the operation station 20 (viewed from below in FIG. 2). In the figure, the second stage 22 and the fifth stage 25 described above can be seen, and the third stage 23 and the fourth stage 24 are located on the front side of these stages 22 and 25. Although not shown in FIG. In the fifth stage 25 of the present embodiment, a tapping operation for the dish 50 is performed. As shown on the right side of FIG. 8, the fifth stage 25 is provided with a compact cylinder 48, and an elevator plate 49 is attached to the compact cylinder 48. In the present embodiment, after the lift plate 49 raises the dish 50 by the compact cylinder 48, the lift plate 49 is sharply lowered to give a shock to the dish 50 by dropping, whereby the dish 50 is tapped.

  Further, in the operation station 20 of the present embodiment, a fiber sensor 52 (FIG. 5) for detecting the presence or absence of the dish table 26 is provided between the fourth stage 24 and the fifth stage 25.

  The automatic cell culture device of the present embodiment having the above configuration is used as follows, for example. First, the case where the medium in the dish 50 is exchanged will be described.

  First, the operation robot 11 takes out the dish table 26 on which the five dishes 50 during cell culture are placed from the incubator 14 (FIG. 1). The removal is performed when the operating robot 11 grips the attachment 29 of the dish table 26. Next, the operation robot 11 conveys the dish table 26 to the operation unit 12 and installs it on the operation station 20. The installation of the dish table 26 is detected by the fiber sensor 52 (FIG. 5).

  Next, in the first stage 21, the suction pad 34 descends to a position substantially at the center of the lid 50 b of the dish 50 by the turning of the turning arm 31, and the lid 50 b is sucked. Whether or not the lid 50b is sucked is determined by whether or not the pressure of the suction pad 34 has become negative. Next, when the suction of the lid 50 b is confirmed, the lid 50 b is opened by the turning of the turning arm 31.

  Subsequently, the ROBO cylinder 41 raises the hit 42. As a result, the dish body 50a is tilted, and the culture medium accumulates on one side of the dish body 50a. In this state, as shown in FIG. 6A, the tip 43 held by the pipette device 54 (FIG. 1) is lowered, and the medium is sucked out. Note that a tip 43 is attached to the pipette device 54 in advance. Next, the ROBO cylinder 41 lowers the claws 42, and the suction pad 34 holding the lid 50b by suction is lowered by the turning of the turning arm 31, and covers the dish body 50a with the lid 50b. Here, whether or not the dish lid 50b is properly closed without riding on the dish body 50a is determined by whether or not the detection plate 36 has contacted the contact switch 35. In this state, the lid 50 b is released from the suction pad 34 by stopping suction at the suction pad 34. The suction pad 34 with the lid 50 b released rises as the turning arm 31 turns.

  Next, the dish table 26 rotates 360/5 degrees in the direction of the arrow 56 in FIG. 2 to move the dish 50 that has been discarded of the culture medium to the second stage 22, and the next dish is moved to the first stage 21. Move up. By repeating such an operation four more times, discarding of the medium of all the dishes 50 is completed. Next, the chip 43 used for discarding the medium is replaced with a new one.

  Next, the operation robot 11 takes out a container containing a new culture medium from the room temperature storage shelf 15 (FIG. 1) or the refrigerated storage, is opened by the capper 53, and is then transported to the temporary station 55 (FIG. 1). The

  Subsequently, in the first stage 21, the lid 50 b of the dish 50 in which the old culture medium has already been discarded is opened in the same manner as described above by turning the turning arm 31. Next, the tip 43 sucks the medium from the container containing the new medium in the temporary station 55 and pours a predetermined amount into the dish 50. Further, the lid 50b is closed in the same manner as described above, and the dish 50 that has been exchanged for a new medium is moved to the second stage 22 by rotating the dish table 26, and the next dish is placed on the first stage 21. Move to. By repeating this operation four more times, the medium exchange of all the dishes 50 is completed. When the medium exchange is completed, the operating robot 11 grips the attachment 29 of the dish table 26 and returns the dish table 26 to the incubator 14.

  Next, the case where subculture operation is performed by the automatic cell culture apparatus of this embodiment is demonstrated. First, in the same manner as described above, the dish table 26 is taken out of the incubator 14 by the operation robot 11 and placed on the operation station 20. Next, in the first stage 21, the lid 50b of the dish 50 is opened by the suction pad 34 in the same manner as described above. Furthermore, after the dish main body 50a is tilted by the rise of the claws 42 in the same manner as described above, the medium is sucked by the tip 43 and discarded. Further, in the same manner as described above, the lid 50 b is closed, and the dish table 26 is rotated to move to the second stage 22, and the next dish is moved onto the first stage 21. By repeating this operation four more times, discarding of the medium of all the dishes 50 is completed. Here, the tip 43 used for sucking the old medium after use is replaced with a new one.

  Next, the container containing the solution for detaching cultured cells containing trypsin and the like is taken out from the refrigerated storage by the operating robot 11, opened by the capper 53, and then transferred to the temporary station 55. The pipetting device 54 sucks the peeling solution with the tip 43. In parallel with this, in the first stage 21, the lid 50b of the dish 50 is opened in the same manner as described above, and after the dish body 50a is placed on the rotating plate 40 by the ascending air cylinder 38 with guide, The dish body 50a is rotated by the rotation of the stepping motor 39. In this state, a peeling solution is poured from the chip 43 to the dish 50. As a result, the stripping solution is evenly distributed on the dish 50. Next, the rotation of the rotating plate 40 stops and descends, and the lid 50b is closed. Subsequently, by the rotation of the dish table 26, the dish 50 containing the peeling solution moves to the second stage 22, and the next dish moves onto the first stage 21. The lid 50b is similarly opened for the next dish 50, and a peeling solution is poured from the chip 43 to the dish 50, and the lid 50b is further closed.

  On the other hand, the dish 50 moved to the second stage 22 by the rotation of the dish table 26 is tilted by the ascending of the swivel arm 46 by the air cylinder 44 and is further subjected to a swing operation by the rotation of the swivel arm 46. Thereby, the peeling solution can be adapted to the cells.

  Further, by repeatedly rotating the dish table 26, the dish 50 containing the peeling solution moves to the fifth stage 25, and is subjected to a tapping operation by an impact caused by the ascending and descending of the elevating plate 49. This further promotes cell detachment.

  Next, the dish 50 moved to the first stage 21 again is poured with physiological saline using a new tip 43 after the lid 50b is opened. At this time, the chip 43 has already been replaced with a new one. Next, after the mixing is performed uniformly by raising and rotating the rotating plate 40, the lid 50b is closed. Further, it is sent to the next second stage 22 by the rotation of the dish table 26. This operation is repeated four more times.

  Next, the lid 50b of the dish 50 returned to the first stage 21 is opened again, and the liquid containing the cells is sucked by the chip 43 replaced with a new one and put into the centrifuge tube. The centrifuge tube is conveyed to the temporary station 55 by the operation robot 11 in advance. Furthermore, by the rotational movement of the dish table 26, the liquid containing the cells of the other dish 50 is transferred to the centrifuge tube. When the cell-containing liquid of all the dishes 50 is collected in the centrifuge tube, the operating robot 11 closes the centrifuge tube with the capper 53, and then installs the centrifuge tube in the centrifuge so that centrifugation is performed. On the other hand, the dish tray 57 on which the emptied dish 50 is placed is removed from the dish table 26 by the operating robot 11 holding and transporting the attachment 60. Next, the dish tray 57 on which the new dish 50 is placed is taken out from the room temperature storage shelf 15 by the operating robot 11 and placed on the dish table 26. Thereby, the exchange of the dish 50 is completed.

  When the centrifugation is completed, the supernatant is discarded by the tip 43 in the temporary station 55, and a new medium is poured.

  Next, in the first stage 21, the lid 50 b of the new dish 50 is opened, and a liquid containing cells is poured from the centrifuge tube by the chip 43. Further, the lid 50 b is closed as described above, and the other dish 50 is moved to the first stage 21 by the rotational movement of the dish table 26. By repeating this operation, after the liquid containing the cells is poured into all of the five dishes 50, the dish table 26 is conveyed to the incubator 14 by the operation robot 11. Further, a dish table 26 on which another new dish 50 is placed is set in the operation station 20, and an operation of pouring a liquid containing cells into the new dish 50 is performed in the same manner as described above. Thus, the passage operation using the automatic cell culture device of the present embodiment is completed.

  In the above description, no operation is performed in the third stage 23 and the fourth stage 24. However, for example, a microscope observation stage for confirming the proliferation of cells in the culture vessel using a microscope may be used. Various stages can be provided depending on the purpose.

  In the above description, the operation station 20 includes the five stages 21 to 25. However, the present invention is not limited to this, and the number of stages can be increased or decreased as necessary.

  Furthermore, although the case where each stage was arrange | positioned circularly was demonstrated above, this invention is not limited to this, For example, it is good also as a structure which arrange | positions each stage in a line and reciprocates a dish table.

  By using the automatic cell culture apparatus of the present invention, various operations in cell culture can be performed quickly, so that the regenerative medicine industry that regenerates cells, tissues, etc. of human skin, cartilage, bone, blood vessels, organs, etc., It can be used in fields such as the bio equipment industry.

It is a top view showing the whole automatic cell culture device concerning one embodiment of the present invention. It is a top view of the operation station in the automatic cell culture apparatus of FIG. (A) is a top view of the dish table used with the automatic cell culture apparatus of FIG. 1, (b) is BB arrow sectional drawing of (a). (A) is a top view of the dish tray used with the dish table of FIG. 3, (b) is the side view. FIG. 3 is a right side view of the operation station of FIG. 2. (A) And (b) is PP sectional view taken on the line in the operation station of FIG. It is a side view showing the 2nd stage 22 along the QQ line of FIG. It is a front view (figure seen from the lower part of Drawing 2) of the operation station of this embodiment.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Case 11 Operation robot 12 Operation part 13 Horizontal axis 14 Incubator 15 Room temperature storage shelf 16 Cell loading / unloading part 17 Compressor 18 Nitrogen gas generator 20 Operation station 21 1st stage 22 2nd stage 23 3rd stage 24 2nd 4 stage 25 5th stage 26 dish table 27 opening window 28 recess 29 attachment 30 block 31 swivel arm 32 stepping motor 33 suction cup unit 34 suction cup pad 35 contact switch 36 detection plate 38 air cylinder with guide 39 stepping motor 40 rotation plate 41 ROBO Cylinder 42 Atari 43 Tip 44 Air Cylinder with Guide 45 Stepping Motor 46 Swivel Arm 47 Ball Plunger 48 Compact Cylinder Die 49 Elevating plate 50 Dish 50a Dish main body 50b Dish lid 51 Fiber sensor 52 Fiber sensor 53 Capper 54 Pipette device 55 Temporary station 57 Dish tray 58 Opening 59 Locking part 60 Attachment 61 Locking member

Claims (10)

An automatic cell culture apparatus equipped with an operation unit for performing a series of operations necessary for cell culture on a plurality of culture vessels,
The operation unit includes an operation station on which a table for holding the plurality of culture vessels is placed, and an operation station having a plurality of stages. At least a part of the plurality of stages is a part of the plurality of operations. The plurality of culture vessels move to the next stage after the operation is performed ,
The operations are performed in parallel in the plurality of stages,
The movement of the plurality of culture vessels to the next stage is performed by rotating the table,
The table holds the plurality of culture vessels arranged in a circle,
The table has a plurality of trays on which culture containers are placed, and the plurality of culture containers are held on the table via the trays,
The robot further comprises an operation robot for performing operations necessary for cell culture in the automatic cell culture apparatus, and the operation robot performs installation of the table in the operation station and removal of the table from the operation station. An automatic cell culture apparatus characterized by performing . The operation in the operation stage, the automatic cell cultivation apparatus according to claim 1, wherein the a closing operation of the lid of the culture vessel. The automatic cell culture apparatus according to claim 1 or 2 , wherein the operation in the operation stage is an inclination operation with respect to the culture vessel. The automatic cell culture device according to any one of claims 1 to 3 , wherein the operation in the operation stage is a pipetting operation on the culture vessel. The automatic cell culture apparatus according to any one of claims 1 to 4 , wherein the operation in the operation stage is a rotation operation with respect to the culture vessel. The automatic cell culture apparatus according to any one of claims 1 to 5 , wherein the operation in the operation stage is a rocking operation of the culture vessel. The operation in the operation stage, the automatic cell cultivation apparatus according to any one of claims 1 to 6, characterized in that said a tapping operation of the culture vessel. The automatic cell culture apparatus according to any one of claims 1 to 7 , wherein the operation in the operation stage is microscopic observation of the culture vessel. Control station for holding a plurality of culture vessels to each of the plurality of stages in a table to be used in the automatic cell cultivation apparatus according to claim 1. The table has a plurality of trays for placing the culture vessel, the plurality of culture vessels, according to claim 9, wherein the table, characterized in that it is held in the table through the tray.

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