JP2019198242A - Pipette for cell culture apparatus - Google Patents

Abstract

To provide a pipette for cell culture apparatus which can prevent contamination in a cell propagation device by preventing splash of liquid.SOLUTION: A pipette for cell culture apparatus comprises a pipette tip 25 detachable from a tip tube 24 and a cap 26 attached to the pipette tip 25. The pipette tip 25 penetrates the cap 26, and is formed such that the cap 26 clogs a mouth part of the cell culture vessel 19 when a tip of the pipette tip 25 is inserted into a cell culture vessel 19.SELECTED DRAWING: Figure 6

Description

  The present invention relates to a pipette for a cell culture device that can prevent contamination in a cell growth device by preventing scattering of liquid.

  In recent years, research on regenerative medicine and cell therapy for treating diseases by transplanting adult stem cells has been actively conducted. In order to perform such treatment, methods for culturing and adjusting adult stem cells in tissues collected from patients have been widely studied. By culturing the cells, it is possible to amplify the number of stem cells that are extremely low in the tissue to the number necessary for treatment.

  Currently, these cell culture devices are only registered with physics and chemistry equipment, and there are no medical equipment, so they must be used and managed by doctors and handled by persons other than doctors. I can't. In this regard, if it is certified as an advanced medical management device (regenerative medical incubator) under the Pharmaceutical Affairs Law by clearing strict safety and ensuring aseptic conditions, use by a doctor is not necessarily required. In other words, if a cell culture device certified as a medical device can be provided, regenerative medicine using the cell culture device can be remarkably spread.

  In addition, in order to popularize regenerative medicine using a cell culture device, a device capable of realizing cell culture safely and at low cost is required. For example, cell culture can be performed safely and at low cost if an apparatus that automatically performs incubation, medium exchange, passage work, and the like can be realized. Moreover, if cell culture of a plurality of subjects can be performed with a single device, efficient cell culture can be executed concurrently.

  With regard to such a cell culture device, Patent Document 1 discloses a configuration in which a robot arranged in the cell culture device operates a pipette device. In the configuration described in Patent Document 1, the robot and the pipette device are controlled such that the container is held by the robot and the sample is sucked or injected into the container by the pipette device fixed to the fixed base. ing.

International Publication No. 2016/170623

By the way, when a cell or culture solution is injected into a container with the pipette device as described above, the dropped liquid may be splashed and scattered outside the container. If dirt is generated in the cell culture device due to the scattered liquid, there is a risk of contamination. Moreover, there is a risk that the scattered cells may develop into a critical problem such as mixing with other specimens.
Then, this invention makes it a subject to provide the pipette for cell culture apparatuses which can prevent the contamination in a cell growth apparatus by preventing scattering of a liquid.

  The present invention has been made in order to solve the above-described problems, and is a pipette device used in a cell growth device to transfer cells to a cell culture container, and includes a tip tube and the tip tube. A detachable tapered pipette tip and a cap attached to the pipette tip, the pipette tip penetrating the cap, and the tip of the pipette tip is inserted into the cell culture container Further, the cap is formed so as to close the mouth of the cell culture container.

  The present invention is as described above, and includes a cap attached to the pipette tip. The pipette tip penetrates the cap, and when the tip of the pipette tip is inserted into the cell culture vessel, the cap It is formed so as to close the mouth. According to such a configuration, since the mouth of the cell culture container is closed, even if the dropped liquid splashes, the liquid does not scatter outside the cell culture container. Contamination in the propagation device can be prevented.

  Further, since the cap is attached to the pipette tip, the cap is automatically changed by changing the pipette tip after use. Therefore, even if liquid adheres to the inside of the cap, there will be no problem in the next pipetting operation.

  In addition, the cap may be attached to the mouth of the cell culture container by screwing. If comprised in this way, a clearance gap will not arise between a cap and the opening | mouth part of a cell culture container, but it can prevent scattering of a liquid reliably.

  In addition, the cap may be provided with a bottom seal portion that is formed of an elastic material and is pressed against the mouth of the cell culture container. If constituted in this way, the bottom seal part is pressed against the mouth part of the cell culture container and elastically deformed, so that the mouth part of the cell culture container can be closed without any gaps, and liquid splash can be reliably prevented. Can do.

  Moreover, you may attach a cap to the taper part of a pipette tip. If comprised in this way, when pressing a cap against the opening | mouth part of a cell culture container, the force which pushes a cap into the back of a taper part will be added. Since the cap is pushed into the back of the taper portion, the pipette tip and the cap are further brought into close contact with each other, so that no gap is generated between the pipette tip and the cap. Therefore, it is possible to reliably prevent the liquid from splashing out of the cell culture container from this gap.

It is a functional block diagram of a cell growth device. It is a figure explaining the internal structure of a cell growth apparatus, Comprising: It is the figure which looked at the cell growth apparatus from the top. It is a figure explaining the internal structure of a cell growth apparatus, Comprising: It is the figure which looked at the cell growth apparatus from the side. It is a figure explaining the connection of a subunit. It is a figure which shows an accommodating part management table, Comprising: (a) The figure of a 1st table, (b) The figure of a 2nd table. It is a figure explaining the action | operation of a pipette apparatus, Comprising: (a) The figure before pipette insertion, (b) The figure after pipette insertion. It is (a) sectional drawing and (b) bottom view of a pipette tip. It is (a) sectional drawing of the pipette tip which concerns on a modification, (b) It is a bottom view.

Embodiments of the present invention will be described with reference to the drawings.
The cell growth device 10 according to this embodiment is for managing the culture of cells (specimens) collected from a patient, for example, and can perform cell growth by fully automated control. As will be described in detail later, the inside of the cell growth apparatus 10 is maintained in a sterile state, and includes a robot apparatus 30 that executes operations associated with culture. The cell growth device 10 receives a sealed container (such as a test tube or a vial) containing somatic cells, automatically executes cell culture, and then discharges the cultured cells in the sealed container.

  This cell growth device 10 can be applied to a system for producing stem cells from blood cells and a system for producing stem cells from other somatic cells. For example, cells cultured by the cell growth device 10 can be transplanted into a patient for cell therapy. In the present embodiment, an example in which the cell growth device 10 is used for cell growth for treatment will be described. However, the present invention is not limited to this, and the cell growth device 10 can be used for culturing any cell.

  As shown in FIGS. 1 to 3, the cell growth device 10 includes an incubator 11, a work chamber 20, a first robot device 30 </ b> A, a second robot device 30 </ b> B, a sterilization device 50, a control device 40, and the like. The

(Incubator 11)
The incubator 11 is a space for culturing cells. The incubator 11 accommodates a cell culture container 19 in which cells and a medium are stored. Although what is used as the cell culture container 19 is not specifically limited, For example, culture tanks, such as a bottle and a bioreactor, a gas-permeable bag, a flask, a petri dish etc. can be used. In the present embodiment, from the viewpoint of ease of handling by the robot apparatus 30, a rectangular tube bottle is used as the cell culture container 19.

  The inside of the incubator 11 is controlled so that the temperature in the cell culture container 19 is kept constant, so that a state suitable for cell growth is maintained. For example, a temperature adjustment device (not shown) that automatically heats or cools the incubator 11 so as to keep the temperature inside the incubator 11 constant is provided inside the incubator 11. In this temperature adjusting device, the output current is controlled in conjunction with the temperature detected by the temperature sensor arranged in the incubator 11. Thereby, the temperature in the incubator 11 is controlled to become the target temperature.

  As shown in FIGS. 2 and 3, the incubator 11 according to the present embodiment is configured by connecting one or more subunits 12. 2 and 3 show a state where three subunits 12 are connected, the number of connections of the subunits 12 can be increased or decreased. That is, the number of connections of the subunits 12 can be arbitrarily set, the number of connections of the subunits 12 may be 2 or less, and the number of connections of the subunits 12 may be 4 or more.

  The subunits 12 according to the present embodiment are standardized so that they all have the same shape and size. And the subunit 12 before a connection is provided with the opening part 12a in the wall part of the both sides of a connection direction, as shown in FIG. By connecting the subunits 12 so that the openings 12a communicate with each other, a plurality of subunits 12 can be coupled in a state where the internal space is communicated.

  Moreover, when connecting the subunit 12 with the working chamber 20 mentioned later, the panel provided with the delivery part 17 mentioned later is engage | inserted by this opening part 12a. Thereby, the subunit 12 and the working chamber 20 can be connected with the delivery part 17 interposed therebetween.

  In addition, the subunit 12 arrange | positioned at the edge on the opposite side to the working room 20 is provided with the opening part 12a to which the subunit 12 and the working room 20 are not connected (rightmost opening part 12a of FIG. 4). . The opening 12a may be closed by fitting a panel or the like. Alternatively, the opening 12a may be closed without being provided from the beginning.

  In addition, the inside of these subunits 12 is formed so that each can be sealed independently, and the incubator room 13 which is the space which can accommodate the cell culture container 19 is comprised. The incubator room 13 is provided for each subunit 12 and is configured such that different environmental conditions can be set for each subunit 12. For this reason, even when the optimum environmental conditions differ depending on the cell type to be cultured, the environmental conditions such as temperature can be set for each subunit 12. And since the incubator room 13 of each subunit 12 is controlled according to the environmental condition set individually, the cell culture which set different environmental conditions can be performed simultaneously. For example, different environmental conditions can be set for one subunit 12 and another subunit 12, and cell culture can be performed under each set environmental condition.

  In the present embodiment, in order to isolate the interior of the subunit 12 (incubator room 13) from each other, a shutter 18 that can be opened and closed is provided at the boundary between the neighboring subunits 12. The shutter 18 is made of a material having high confidentiality and heat insulation, and the inside of the subunit 12 can be sealed by closing the shutter 18. By isolating the inside of the subunit 12 with the shutter 18, different culture environments can be realized for each subunit 12. Further, when executing the sterilization process described later, the sterilization process can be executed for each subunit 12 by closing the shutter 18 and isolating the interior of the subunit 12.

  As shown in FIGS. 2 and 3, the subunit 12 includes a plurality of rows of accommodating portions 14 for accommodating the cell culture containers 19. Each accommodating portion 14 is configured to accommodate one cell culture container 19. The accommodating portions 14 are arranged in a plurality of rows in the horizontal direction and in a plurality of stages in the vertical direction. Further, each subunit 12 is provided with accommodating portions 14 having the same number of rows and the same number of stages. In the present embodiment, as shown in FIG. 3, three rows of storage portions 14 are provided in the horizontal direction and four levels in the vertical direction, respectively.

  In the present embodiment, one cell culture container 19 can be accommodated in each accommodating part 14, but the present invention is not limited thereto, and a plurality of cell culture containers 19 may be accommodated in each accommodating part 14. Further, in the present embodiment, the accommodating portions 14 having three rows in the horizontal direction and four steps in the vertical direction are provided, but the present invention is not limited to this, and the number of rows and the number of stages of the accommodating portions 14 can be arbitrarily set. .

  Here, the above-mentioned accommodating part 14 is provided in the both sides of each subunit 12, as shown in FIG. That is, in the present embodiment, three rows of storage portions 14 in the horizontal direction and four stages in the vertical direction are provided on both sides of each subunit 12. Thus, by providing the accommodating portions 14 on both sides of the subunit 12, the area of the accommodating portion 14 can be increased. As will be described in detail later, a second robot device 30B is provided between the storage units 14 on both sides, and the second robot device 30B executes processing for the storage unit 14. For this reason, although the accommodating part 14 needs to face the 2nd robot apparatus 30B, providing the accommodating part 14 on both sides can increase the area of the accommodating part 14 which can face the 2nd robot apparatus 30B. it can.

  Further, as shown in FIG. 3, the subunit 12 according to the present embodiment can be connected to the subunit 12 in the arrangement direction of the accommodating portions 14 (the row direction of the accommodating portions 14, that is, the left-right direction in FIG. 3). It has become. For this reason, it is configured such that the number of rows in the horizontal direction of the accommodating portion 14 is increased or decreased by increasing or decreasing the number of connections of the subunits 12. For example, when the number of subunits 12 is one, the storage sections 14 are arranged in three rows (four stages). When two subunits 12 are connected, the storage section 14 is arranged in six rows (four stages). When three are connected, it is configured so as to form nine rows (four stages) of accommodating portions 14.

As described above, the cell growth device 10 according to the present embodiment can set different environmental conditions depending on the plurality of subunits 12, and includes a plurality of storage units 14. Can grow in parallel.
(Workroom 20)
The work room 20 is an area for performing work related to a preparation process before incubation. The work chamber 20 is disposed adjacent to the incubator 11 and is coupled to the end subunit 12 of the plurality of subunits 12 constituting the incubator 11. The inside of the work chamber 20 forms a sealed space isolated from the subunit 12.

  As shown in FIGS. 2 and 3, a delivery unit 17 is arranged between the subunit 12 at the end adjacent to the working chamber 20 and the working chamber 20. The delivery unit 17 is a small room for delivering the cell culture container 19 and the like, and is configured so that the inside becomes a sealed space. The transfer section 17 is provided with a sealing door on both the working chamber 20 side and the subunit 12 side, and these two sealing doors are controlled so as not to be opened simultaneously. In other words, when delivering the cell culture container 19 or the like via the delivery unit 17, the sealed door is controlled so that only one of them is opened.

  For example, when the cell culture container 19 is delivered from the working chamber 20 to the subunit 12, first, the hermetic door on the working chamber 20 side is opened. 19 is placed. Then, after the closed door on the side of the working chamber 20 is closed, the closed door on the side of the subunit 12 is opened, so that the second robot device 30B described later receives the cell culture container 19 placed on the transfer unit 17. Thereafter, the sealing door on the subunit 12 side is closed, and the delivery is completed. By such control, the working chamber 20 and the subunit 12 are not in communication with each other, and are always kept in an isolated state.

  Further, as shown in FIGS. 2 and 3, the work chamber 20 is provided with a work table 21 for performing work such as medium replacement. On the work table 21, a plurality of devices for performing operations such as medium exchange, for example, devices such as a pipette device 22 and a centrifuge 28 are arranged.

  The pipette device 22 is a device that sucks and injects a sample such as a culture medium by a preset volume. The pipette device 22 according to the present embodiment is controlled by the control device 40 so that suction and injection processing can be executed. The pipette device 22 may be used with a known micropipette fixed to a table, or a robot incorporating a micropipette may be used.

  The centrifuge 28 is a device for generating a centrifugal force to separate necessary cells. For example, after the liquid containing the cells is applied to the centrifuge 28, the supernatant is discarded, and the cells settled on the bottom are sucked out by the pipette device 22.

(First robotic device 30A)
The first robot device 30A is a robot device 30 that is in charge of a preparatory step before incubation. The first robot apparatus 30 </ b> A is a stationary apparatus disposed in the work chamber 20. The first robot apparatus 30A is arranged so as to face the work table 21 described above, and performs operations such as medium exchange on the work table 21 using various devices and instruments.

  The first robot apparatus 30A is a so-called single-arm robot capable of handling the cell culture container 19 with a robot arm. Specifically, the first robot apparatus 30 </ b> A includes a base 31 fixed to the work chamber 20 and an arm 32 fixed on the base 31.

  The base 31 is fixed to the floor surface of the work chamber 20 with, for example, anchor bolts. The base 31 may be fixed to the ceiling surface or the side surface instead of the floor surface of the work chamber 20.

  Moreover, the arm 32 according to the present embodiment is configured as an arm having six joint portions and having six degrees of freedom. A pair of claw portions 33 is provided at the tip of the arm 32, and the pair of claw portions 33 are configured to be able to grip the cell culture container 19 and various instruments by moving in a direction away from each other. Yes.

(Second robot device 30B)
The second robot apparatus 30 </ b> B is a robot apparatus 30 that is responsible for taking in and out the cell culture container 19 to and from the incubator 11. The second robot apparatus 30B is disposed inside the incubator 11.

  Note that a continuous rail 36 is provided on the floor surface of the incubator 11 so as to cross the subunit 12 as shown in FIG. The second robot device 30 </ b> B can move in the lateral direction along the rail 36 by traveling on the rail 36. The second robot device 30B can move freely across the connected subunits 12 by moving along the rail 36.

  In addition, as already demonstrated, the accommodating part 14 inside the incubator 11 is arrange | positioned on both sides on both sides of the rail 36, as shown in FIG. For this reason, when the second robot apparatus 30 </ b> B moves along the rail 36, the second robot apparatus 30 </ b> B moves in the lateral direction along the extending surface of the housing portion 14. For this reason, the second robot device 30 </ b> B can perform the processing in a face-to-face manner when processing is performed on any of the accommodating portions 14. In addition, since the accommodating portions 14 are provided on both sides of the second robot device 30B, the area of the accommodating portion 14 can be increased while shortening the moving distance (the length of the rail 36) of the second robot device 30B as much as possible. It has become.

  By the way, in the incubator 11 performing the culture operation, the subunits 12 are shielded by the shutter 18. For this reason, when the second robot apparatus 30B moves across the subunit 12, the shutter 18 is opened and closed. That is, when the second robot apparatus 30B moves across the subunit 12, the shutter 18 is first opened, and after the second robot apparatus 30B moves across the subunit 12, the shutter 18 is closed. ing. The opening / closing process of the shutter 18 may be executed by a control signal from the control device 40, or the second robot device 30B may operate an opening / closing button (not shown) of the shutter 18.

  The second robot apparatus 30B is a so-called single-arm robot that can handle the cell culture container 19 with a robot arm. Specifically, the second robot apparatus 30 </ b> B includes a base 31 configured to run on the rail 36 and an arm 32 fixed on the base 31. In the present embodiment, the rail 36 is provided on the floor surface of the subunit 12, but not limited to this, the rail 36 may be provided on the ceiling surface or side surface of the subunit 12. In the present embodiment, the base 31 runs on the rail 36. However, the present invention is not limited to this, and the second robot device 30B is provided with rails by providing wheels and a caterpillar device on the base 31. You may enable it to drive | work a plane irrespective of 36.

  Moreover, the arm 32 according to the present embodiment is configured as an arm having six joint portions and having six degrees of freedom. A pair of claw portions 33 is provided at the tip of the arm 32, and the pair of claw portions 33 are configured to be able to grip the cell culture container 19 and various instruments by moving in a direction away from each other. Yes.

(Sterilizer 50)
The sterilizer 50 is a device that executes sterilization / sterilization processing in the incubator 11 and the work chamber 20. Although not particularly illustrated, the sterilization apparatus 50 according to the present embodiment includes a PSA-type oxygen generator and a discharge device.

  The PSA oxygen generator is an apparatus that separates air in the atmosphere into nitrogen and high-concentration oxygen, and a known apparatus can be used. The oxygen generated by the PSA oxygen generator is sent to the discharge device for use.

  The discharge device is a device that generates ozone by applying a voltage between electrodes to generate discharge, and a known device can be used. The discharge device includes an air flow path to which oxygen generated by a PSA oxygen generator is supplied, generates discharge plasma on the air flow path, and converts ozone from oxygen passing through the air flow path to ozone. Is configured to generate

  The ozone gas generated by the sterilizer 50 is supplied into the incubator 11 and the work chamber 20 through a pipe. That is, as shown in FIG. 3, a supply port 51 connected to the sterilizer 50 is provided in the ceiling of each incubator room 13 and work chamber 20. When the sterilizer 50 generates ozone gas, the ozone gas is configured to be ejected from the supply port 51 into each incubator room 13 or the work chamber 20. Thus, by supplying ozone gas and filling the sealed space with ozone gas, sterilization and sterilization are performed.

  Note that the sterilization apparatus 50 according to this embodiment can supply ozone gas to each incubator room 13 and the work room 20 individually. That is, it is possible to supply ozone gas only to one or more spaces selected from among the plurality of incubator rooms 13 and the work room 20. And as mentioned above, since each incubator room 13 (subunit 12) and the working room 20 are formed so as to be able to be sealed independently, only the space supplied with ozone gas is selectively sterilized and sterilized. Can do.

(Control device 40)
The control device 40 is for controlling various devices such as the robot device 30 described above. The control device 40 is configured around a CPU and includes a ROM, a RAM, an input / output port, and the like. Further, various input devices and output devices are controlled by an operating system stored in the ROM.

  In addition, as an input device connected to the control device 40, although not particularly illustrated, for example, an input device (one or more) such as a liquid crystal touch panel, a keyboard, a mouse, a button, and a barcode reader can be provided. When these input devices accept an operation from the user, the control device 40 controls the operation of the cell growth device 10 according to the operation content.

  Further, as an output device connected to the control device 40, as shown in FIG. 1, the subunit 12 of the incubator 11, the shutter 18, the pipette device 22, the first robot device 30A, the second robot device 30B, sterilization, and the like. A device 50 or the like is provided.

  As shown in FIG. 1, the control device 40 is configured such that the CPU executes a program stored in the ROM, so that the schedule management unit 41, the first robot control unit 42, the second robot control unit 43, and the arrangement management unit 46. , Function as a subunit state management unit 47. Note that the control device 40 is not limited to these functions, and may include other functions.

(Schedule management unit 41)
The schedule management unit 41 is for managing a schedule of a process related to cell proliferation. For example, the ID of the cell culture container 19 is stored, and the schedule until the cell culture container 19 finishes cell growth is managed. The schedule managed by the schedule management unit 41 includes the timing of medium replacement and passage operation. For this reason, when it is the timing of medium exchange or passage operation for a certain cell culture container 19, the schedule management unit 41 issues a command to the first robot control unit 42 and the second robot control unit 43 described later, The first robot device 30A and the second robot device 30B are used to perform medium exchange and passage operation.

  In addition, the schedule management unit 41 executes the synchronization process when each apparatus needs to perform the process in synchronization. For example, when the cell culture container 19 is delivered from the first robot device 30A to the second robot device 30B, the first robot device 30A waits for a predetermined operation (first robot controller). Waiting for a work completion signal from 42), the second robot apparatus 30B is caused to execute a predetermined operation (a work start signal is transmitted to the second robot control unit 43). By controlling the processing timing in this way, a series of processes can be executed without delay.

(First robot controller 42)
The first robot control unit 42 is for controlling the first robot apparatus 30A. The first robot control unit 42 performs a predetermined operation based on a control signal from the schedule management unit 41. Programs related to the operation of the first robot apparatus 30A are registered as several processing units. The first robot control unit 42 reads and executes the processing unit selected by the schedule management unit 41, thereby operating the first robot apparatus 30A with some predetermined patterns. For example, the first robot device 30A is caused to perform operations such as suction processing or injection processing using the pipette device 22 and cell extraction using the centrifuge 28.

(Second robot control unit 43)
The second robot control unit 43 is for controlling the second robot device 30B. The second robot control unit 43 executes a predetermined operation based on the control signal from the schedule management unit 41. Programs related to the operation of the second robot apparatus 30B are registered as several processing units. The second robot control unit 43 can operate the second robot apparatus 30B with some predetermined patterns by reading and executing the processing unit selected by the schedule management unit 41. For example, the second robot apparatus 30 </ b> B is caused to perform an operation such as taking the cell culture container 19 into and out of the incubator 11.

(Placement management unit 46)
The arrangement management unit 46 is for managing which storage unit 14 of the incubator 11 stores which cell culture container 19 is stored. The arrangement management unit 46 according to the present embodiment manages the row and column numbers of the storage unit 14 and the ID of the cell culture container 19 stored in the storage unit 14 in association with each other. For this reason, the accommodating part 14 and the cell culture container 19 are memorize | stored by the one-to-one relationship, and it is comprised so that the mixing of the cell culture container 19 may not occur.

  For example, the arrangement management unit 46 manages the position of the storage unit 14 using a storage unit management table as shown in FIG. The storage unit management table stores the row and column numbers of the storage unit 14 and the ID of the cell culture container 19 stored in the storage unit 14. In the present embodiment, since the accommodating portions 14 are provided on both sides of the second robot device 30B, the accommodating portion 14 on one side (for example, the right side when viewed from the work chamber 20) of the second robot device 30B is provided. Two storage unit management tables including a first table for management and a second table for managing the storage unit 14 on the other side (for example, the left side when viewed from the working chamber 20) of the second robot apparatus 30B. Is provided. The placement management unit 46 can manage which cell culture container 19 is stored in which storage unit 14 by referring to the storage unit management table.

  For example, according to the storage unit management table shown in FIG. 5, it can be seen that the cell culture container 19 with ID “ABB” is stored in the second row and second row of the storage unit 14 on the right side. Further, it can be seen that the cell culture container 19 with ID “BDA” is accommodated in the first row and the fourth row of the accommodation portion 14 on the left side.

  In this example, the ID of the cell culture container 19 is three alphabetic characters, but what ID is used can be freely set. For example, in consideration of ease of handling on a computer, an 8-bit numerical value or the like may be used, or a character string such as a subject's name may be used giving priority to human visibility.

  Thus, since the arrangement management unit 46 manages the position of the storage unit 14 in the vertical and horizontal directions (XY directions), even if the storage unit 14 increases in the vertical or horizontal direction, the storage unit 14 is processed in the same manner. Can be managed. That is, even if the number of connected sub-units 12 increases, the number of columns shown in FIG. 5 only increases, so that the position of the accommodating portion 14 can be managed without increasing the type of management table or rewriting the program. can do.

(Subunit state management unit 47)
The subunit state management unit 47 is for managing the state of the subunit 12.
For example, the subunit state management unit 47 can manage different environmental conditions for each subunit 12. Specifically, when the user sets the environmental condition of each subunit 12 using the input device, this environmental condition is transmitted to the subunit state management unit 47. The subunit state management unit 47 manages the temperature and pH of each subunit 12 so as to satisfy the environmental conditions while referring to the set environmental conditions. As a result, cell culture can be performed simultaneously in parallel under independent environmental conditions for each subunit 12.

(Basic procedure for cell growth)
Cell growth using the cell growth device 10 according to the present embodiment is executed in the following procedure.
First, a tissue is collected from a patient by a doctor, thereby obtaining cells to be cultured. The cells are stored in a sealed container.

  Then, the person in charge of the culture work activates the cell growth device 10 and inputs a command to start culture. At this time, the operator may input parameters necessary for culture. For example, a patient identification number, information (volume) about the cell culture container 19 and the like may be input. In order to facilitate the input of these parameters, a barcode may be attached to the cell culture container 19. By reading this bar code with a bar code reader provided in the cell growth device 10, parameters can be easily input.

  When the cell growth device 10 receives the culture start command, the control device 40 checks the available space in the incubator 11. If it is determined that there is sufficient space, a pass box (not shown) for delivering the container to the cell growth device 10 is activated. The person in charge puts the sealed container containing the cells and the cell culture container 19 into the pass box. The cell growth device 10 automatically pulls back the pass box, thereby taking the sealed container and the cell culture container 19 into the cell growth device 10.

At this time, the cell growth device 10 may notify the worker in charge of replenishment of various consumables. For example, the operator may be notified to put a container filled with a culture medium, a container filled with a growth factor, and the like into the pass box. And you may make it wait for the start of culture | cultivation until a required consumable is provided.
When the sealed container and the cell culture container 19 are taken in, processing by the robot device 30 is executed.

  That is, the sealed container and the cell culture container 19 are first fed into the working chamber 20, and a pre-incubation preparatory step is executed by the first robot apparatus 30A. Specifically, a predetermined amount of culture medium or growth factor is supplied to the cell culture container 19. Further, the contents of the sealed container are put into a centrifuge 28, and the cells used for culture are taken out. The cells are supplied to the cell culture container 19 using the pipette device 22.

  The cell culture container 19 supplied with the cells in this way is delivered from the work chamber 20 to the incubator 11. The cell culture container 19 delivered to the incubator 11 is accommodated in the predetermined accommodating portion 14 by the second robot apparatus 30B. The cell culture container 19 accommodated in the accommodating part 14 is stored in the incubator 11 until the set culture period (for example, several days) elapses. In the incubator 11, cell growth proceeds by maintaining the cell culture container 19 at a constant temperature.

When the culture period has elapsed and the cells necessary for the treatment have been prepared, the cell growth apparatus 10 notifies the worker in charge of the completion of the culture. When the worker who has received the notification requests the cell growth apparatus 10 to receive cells, the second robot apparatus 30B takes out the cell culture container 19 from the incubator 11. The extracted cell culture container 19 is delivered to the first robot apparatus 30A. The first robot apparatus 30A loads the cells in the cell culture container 19 into a syringe. This syringe is discharged to the outside of the cell growth device 10 through a pass box.
Thus, the cell collect | recovered from the cell growth apparatus 10 is inject | poured and used for a patient's body for a cell therapy, for example.

(Procedures such as medium exchange)
In the cell growth device 10 of the present embodiment, medium replacement and passage operation, which are culture operations necessary for cell culture, can be performed automatically. For example, medium exchange is performed as follows.

  First, when the schedule management unit 41 detects the presence of the cell culture container 19 that requires medium replacement by checking the schedule, it instructs the second robot control unit 43 to take out the cell culture container 19.

  Receiving the command, the second robot control unit 43 operates the second robot device 30B to take out the cell culture container 19 accommodated in the accommodation unit 14 of the incubator 11. That is, first, the position of the cell culture container 19 to be taken out is grasped by making an inquiry to the arrangement management unit 46. Then, the operation of the second robot device 30B is controlled so that the cell culture container 19 at the grasped position is taken out.

Under the control of the second robot control unit 43, the second robot apparatus 30 </ b> B takes out the cell culture container 19 from the storage unit 14 and places the taken cell culture container 19 on the delivery unit 17. When the cell culture container 19 is placed, the second robot control unit 43 notifies the schedule management unit 41 of the completion of removal of the cell culture container 19.
The schedule management unit 41 that has received the notification of the completion of removal of the cell culture container 19 instructs the first robot control unit 42 to execute the medium replacement process.
Upon receiving the command, the first robot control unit 42 causes the first robot apparatus 30A to execute the following processing, for example.

That is, first, the first robot apparatus 30 </ b> A receives the cell culture container 19 placed on the transfer unit 17 and moves it to the work table 21. Then, the lid of the cell culture container 19 is opened. Subsequently, using the pipette device 22, the medium in the cell culture container 19 is aspirated and discarded, and a new medium is supplied to the cell culture container 19. Finally, the lid of the cell culture container 19 is closed, and the cell culture container 19 with the lid closed is placed on the delivery unit 17.
When the placement of the cell culture container 19 is completed, the first robot control unit 42 notifies the schedule management unit 41 of completion of the medium replacement process.
The schedule management unit 41 that has received the notification of the completion of the medium exchange process instructs the second robot control unit 43 to store the cell culture container 19.

  Receiving the command, the second robot control unit 43 operates the second robot device 30B and accommodates the cell culture container 19 in the original position. Thus, the medium exchange is completed.

(About the pipette device 22)
As shown in FIG. 6, the pipette device 22 according to the present embodiment includes a drive unit 23, a tip tube 24, a pipette tip 25, and a cap 26.

The drive unit 23 is a part having a built-in mechanism for operating the pipette device 22. The drive unit 23 according to the present embodiment incorporates a cylinder and a piston capable of reciprocating in the cylinder, although not particularly illustrated. When the controller 40 commands the execution of the suction operation, the drive unit 23 performs an operation of pulling up the piston, thereby generating a negative pressure in the distal end tube 24 and sucking up the liquid. Moreover, when execution of injection | pouring operation is commanded by the control apparatus 40, the drive part 23 will perform the operation | movement which pulls down a piston, and this is comprised so that the sucked-up liquid may be discharged | emitted.
The tip tube 24 is a tubular member that protrudes downward from the tip of the drive unit 23. The inside of the tip tube 24 communicates with the cylinder of the drive unit 23.

  The pipette tip 25 is a tubular member that can be attached to and detached from the tip of the tip tube 24. The pipette tip 25 is replaced every time a pipetting operation (aspiration and injection) is performed by the pipette device 22. The pipette tip 25 has a tapered shape and includes an attachment portion 25a and a taper portion 25b as shown in FIG.

  The attachment portion 25 a is a cylindrical portion that is attached to the distal end tube 24. The attachment portion 25a is formed on the proximal end side (thick end portion) of the pipette tip 25, and the pipette tip 25 can be attached to the distal end tube 24 by pressing the attachment portion 25a against the distal end tube 24. ing.

  The tapered portion 25b is a conical portion formed so as to be continuous with the attachment portion 25a. An opening for sucking or injecting liquid is formed at the tip of the taper portion 25b.

  The cap 26 is a member attached to the pipette tip 25. The cap 26 according to the present embodiment is a short cylindrical lid as shown in FIG. As shown in FIG. 6B, the cap 26 is formed so as to close the mouth of the cell culture container 19 when the tip of the pipette tip 25 is inserted into the cell culture container 19.

  A hole 26 a is formed through the center of the cap 26. The pipette tip 25 is inserted into the hole 26 a and penetrates the cap 26. At this time, the tapered portion 25 b (conical surface) of the pipette tip 25 is attached so as to face the inside of the hole portion 26 a of the cap 26.

  An annular seal portion 26c is attached to the inner periphery of the hole portion 26a. The seal portion 26 c is formed of an elastic member such as rubber and is in close contact with the taper portion 25 b of the pipette tip 25. By providing the seal portion 26 c, no gap is generated between the hole portion 26 a of the cap 26 and the tapered portion 25 b of the pipette tip 25.

Further, on the inner peripheral surface of the cylinder of the cap 26, a female thread portion 26b is formed as shown in FIG. The female screw portion 26b is formed so as to be screwed to a male screw portion 19a (see FIG. 6A) provided on the outer periphery of the mouth of the cell culture container 19. For this reason, the internal thread part 26b is screwed into the external thread part 19a by pressing the cap 26 against the mouth of the cell culture container 19 and rotating it. Thus, the cap 26 can be screwed into the mouth of the cell culture container 19 and attached.
The above-described suction and injection processing by the pipette device 22 is executed in the following procedure.

  Although not particularly illustrated, a pipette rack for storing the pipette tips 25 is provided on the working chamber 20. In this pipette rack, a plurality of pipette tips 25 with caps 26 attached are stored. When the pipette device 22 performs suction and injection, the first robot device 30A takes out the pipette tip 25 to be used from the pipette rack, and presses and attaches the mounting portion 25a of the pipette tip 25 taken out to the tip tube 24.

  Next, the first robot apparatus 30 </ b> A grips a container such as the cell culture container 19 to be aspirated and moves it to the tip position of the pipette tip 25. When the tip of the pipette tip 25 is fixed so as to be immersed in the sample or the like, the drive unit 23 is operated to perform a suction operation. Thereby, the liquid is sucked into the pipette tip 25. When the contents of the cell culture container 19 are difficult to suck in a small amount, the cell culture container 19 held by the first robot apparatus 30A may be tilted to suck the bottom corner.

  Next, the first robot apparatus 30 </ b> A grips a container such as the cell culture container 19 to be injected and moves it to the tip position of the pipette tip 25. Then, an operation is performed so that the tip of the pipette tip 25 is inserted into the cell culture container 19. When the cap 26 is moved to a position where it abuts against the mouth of the cell culture container 19, the pipette tip 25 is rotated together with the tip tube 24 while the cap 26 is abutted against the mouth of the cell culture container 19. By this process, as shown in FIG. 6B, the cap 26 is screwed onto the mouth of the cell culture container 19 and attached. When the mouth of the cell culture container 19 is thus closed, the driving unit 23 is operated to perform the injection operation. As a result, the liquid in the pipette tip 25 is discharged to the cell culture container 19.

  When suction and injection are completed as described above, the first robotic device 30A removes the pipette tip 25 from the tip tube 24. The removed used pipette tips 25 are discarded in a dust box (not shown) arranged below the work table 21.

(Summary)
The present embodiment is as described above, and includes a cap 26 attached to the pipette tip 25. The pipette tip 25 penetrates the cap 26, and the tip of the pipette tip 25 is inserted into the cell culture container 19. The cap 26 is formed so as to close the mouth of the cell culture container 19. According to such a configuration, since the mouth portion of the cell culture container 19 is closed, even if the dropped liquid splashes, the liquid does not scatter outside the cell culture container 19. Contamination in the cell growth device 10 can be prevented.

  Further, since the cap 26 is attached to the pipette tip 25, the cap 26 is automatically exchanged by exchanging the pipette tip 25 after use. Therefore, even if liquid adheres to the inside of the cap 26, there will be no problem in the next pipetting operation.

  Further, the cap 26 is attached to the tapered portion 25b of the pipette tip 25. For this reason, when the cap 26 is pressed against the mouth of the cell culture container 19, a force to push the cap 26 into the back of the tapered portion 25b is applied. By pushing the cap 26 into the back of the taper portion 25b, the pipette tip 25 and the cap 26 are further brought into close contact with each other, so that no gap is generated between the pipette tip 25 and the cap 26. Therefore, it is possible to reliably prevent liquid from splashing out of the cell culture container 19 from this gap.

  Further, the cap 26 can be screwed onto the mouth of the cell culture container 19 and attached. For this reason, there is no gap between the cap 26 and the mouth of the cell culture container 19, and the liquid can be reliably prevented from scattering.

  In the above-described embodiment, the example in which the cap 26 is screwed and attached to the mouth of the cell culture container 19 has been described, but the embodiment of the present invention is not limited thereto. For example, a cap 26 as shown in FIG. 8 may be used.

  The cap 26 shown in FIG. 8 is a disc-shaped lid. The cap 26 is formed so as to close the mouth of the cell culture container 19 when the tip of the pipette tip 25 is inserted into the cell culture container 19.

  A hole 26 a is formed through the center of the cap 26. The pipette tip 25 is inserted into the hole 26 a and penetrates the cap 26. At this time, the pipette tip 25 has penetrated the hole 26a in the taper taper part 25b (conical surface). An annular seal portion 26c is attached to the inner periphery of the hole portion 26a. The seal portion 26 c is formed of an elastic member such as rubber and is in close contact with the taper portion 25 b of the pipette tip 25. By providing the seal portion 26 c, no gap is generated between the hole portion 26 a of the cap 26 and the tapered portion 25 b of the pipette tip 25.

  Further, the bottom surface of the cap 26 is provided with a bottom surface seal portion 26d formed of an elastic material such as rubber and pressed against the mouth of the cell culture container 19. The bottom seal portion 26d is provided in an annular shape so as to contact the entire circumference of the mouth of the cell culture container 19.

  According to such a configuration, the bottom seal portion 26d is pressed against the mouth portion of the cell culture container 19 and elastically deformed, so that the mouth portion of the cell culture container 19 can be closed without a gap, and the liquid can be reliably scattered. Can be prevented.

DESCRIPTION OF SYMBOLS 10 Cell proliferation apparatus 11 Incubator 12 Subunit 12a Opening part 13 Incubator room 14 Storage part 17 Delivery part 18 Shutter 19 Cell culture container 19a Male thread part 20 Work room 21 Work table 22 Pipette device 23 Drive part 24 Tip tube 25 Pipette tip 25a Mounting portion 25b Taper portion 26 Cap 26a Hole portion 26b Female thread portion 26c Seal portion 26d Bottom seal portion 28 Centrifuge 30 Robot device 30A First robot device 30B Second robot device 31 Base 32 Arm 33 Claw portion 36 Rail 40 Control Device 41 Schedule Management Unit 42 First Robot Control Unit 43 Second Robot Control Unit 46 Arrangement Management Unit 47 Subunit Status Management Unit 50 Sterilizer 51 Supply Port

Claims (4)

A pipetting device used in a cell growth device to transfer cells to a cell culture vessel,
A tip tube;
A tapered pipette tip that can be attached to and detached from the tip tube;
A cap attached to the pipette tip;
With
The pipette tip penetrates the cap;
A pipette for a cell culture apparatus, wherein the cap is formed so as to close the mouth of the cell culture container when the tip of the pipette tip is inserted into the cell culture container.   The pipette for a cell culture device according to claim 1, wherein the cap can be screwed onto and attached to the mouth of the cell culture container.   The pipette for a cell culture device according to claim 1, wherein the cap includes a bottom seal portion formed of an elastic material and pressed against the mouth of the cell culture container.   The pipette for a cell culture device according to any one of claims 1 to 3, wherein the cap is attached to a tapered portion of the pipette tip.

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