JP2012147685A - Cell culture treatment system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To solve problems that exclusive devices capable of performing various cell culture processes are large, complicated, and high costs.SOLUTION: There is provided a cell culture treatment system, in which treatment devices bearing necessary processes in response to cells-tissues are prepared; the treatment devices connect closed spaces capable of being sterilized or decontaminated with detachable means; and necessary treatment devices can be chosen, combined and connected, while keeping a closing state and a sterilizing state.

Description

  The present invention relates to an apparatus for culturing cells and the like optimal in the fields of medicine, biology, drug discovery, pharmacy and the like. In particular, the present invention relates to a cell culture treatment system for cells, tissues, organs and the like (hereinafter referred to as cell tissues) that perform cell proliferation and cell organization for regenerative medicine or biopharmaceutical production.

  In recent years, expectations for regenerative medicine that uses human or animal cells to repair lost or degraded tissues and organs have increased, and research and development on the proliferation and organization of somatic cells and somatic stem cells has increased. Further progress has been made in ES cell research and technology development to create and proliferate iPS cells.

In order to culture and further organize cells, tissues, and organs used in these treatments, a cell culture treatment system that solves problems such as safety, high quality, high throughput, and total manufacturing cost reduction is desired. It is rare.

When cells are proliferated or organized for the purpose of transplantation and treatment in human or animal bodies, such as regenerative medicine, proliferation, differentiation or organization must be effectively and efficiently performed. In addition, since it is transplanted and treated in the body of a human or animal, ensuring safety is important.
One of the safety needs to prevent foreign substances and dust from entering, as well as prevent invasion of various bacteria and maintain sterility. Second, especially when the patient's own cells are cultured (autologous culture), it is necessary to prevent contamination by other people, other types of cells, non-autologous cells, viruses derived from these cells, bacteria, etc. .
In order to maintain safety, a clean room, which is a facility with very large facility costs and maintenance costs, is used, but this significantly increases the cost of cultured cells and tissues.
Furthermore, in a cell culture device that automates culture, GMP performs the specified work procedure and work contents in the same way as culturing by human work, confirming that it is actually performed, It is necessary to record and store it, and if these are performed by taking advantage of the characteristics of the mechanical device, the culture and its guarantee can be performed more efficiently and reliably.
On the other hand, for the culture, separation of cells from collected tissues, isolation of target cells from the separated cells, primary culture of cells, amplification culture / passage culture, organization, packaging of cells / tissues, etc. However, depending on the cells or tissues to be cultured, some of these steps may not be necessary, and those steps may be switched in order, some may not be necessary, May repeat several times.
Further, the time required to perform each process varies depending on the process.

JP2008-054690 JP2004-350640

  The problem to be solved is that a dedicated device capable of performing various cell culture processes is large and complicated, and the cost is high.

  The cell culture treatment system of the present invention is a cell culture treatment system in which a plurality of treatment devices including a first treatment device and a second treatment device are detachably connected to each other via attachment / detachment means. Opens and closes a base, a robot that transports the culture plate and the like provided on the base, a housing that blocks the movable atmosphere of the robot from outside air, and an opening that passes the culture plate and the like inside and outside the housing A cell comprising: an opening and closing door; a decontamination device that supplies a decontamination gas; and an attaching / detaching unit that connects to the second processing device, wherein the second processing device includes a moving unit at a lower part. Culture treatment system.

  Such a cell culture treatment system can easily increase or decrease the number of treatment devices according to the type, throughput, and production volume of the culture process.

In the cell culture treatment system of the present invention, when the second treatment apparatus is connected to the housing of the first treatment apparatus by the attachment / detachment means, the decontamination apparatus of the first treatment apparatus via the attachment / detachment means. A pipe for supplying a decontamination gas to the second processing apparatus is connected.

  Thereby, even in the processing apparatus detached by the attaching / detaching means, the decontamination gas can be supplied to the inside of the second processing apparatus when connected to the base of the transport apparatus. It is possible to prevent contamination of bacteria, foreign matters, and other cells that adversely affect cell culture inside the processing apparatus.

  The cell culture processing system of the present invention includes a plurality of the attaching / detaching means, and the plurality of second processing apparatuses can be selectively attached to / detached from the casing of the first processing apparatus via the attaching / detaching means. It is characterized by that.

Thereby, the combination of each processing apparatus can be changed by increasing / decreasing a 2nd processing apparatus via an attachment / detachment means, and it can respond to culture | cultivation processes, such as many kinds of cells and tissues. In addition, each processing device has a unique ID that can be identified, and can be operated by attaching or detaching to any port of the attaching / detaching means. The operation log and the communication log measurement result can be uniquely identified. By transmitting and receiving as an electrical signal to the integrated control device together with the ID, it is possible to manage so that the cell as a specimen is not mixed with other cells.

The cell culture treatment system of the present invention includes a plurality of the first treatment devices.

The cell culture treatment system according to claim 2, wherein the outside of the door is decontaminated by providing a gas supply port from the decontamination device outside the door. Decontamination and sterilization of places that may be contaminated with foreign matter exposed to the external atmosphere before connection after the cell processing operation is performed after the second processing device and the transport device are connected. can do. For this reason, the risk of introducing foreign substances into each processing apparatus is reduced.

  The cell culture treatment system of the present invention has the advantage that the combination of the treatment devices can be changed by increasing or decreasing the treatment devices via the attaching / detaching means, and can cope with the cultivation processes of many types of cells, tissues, and the like. .

FIG. 1 is a plan view showing a cell culture treatment system. FIG. 2 is a plan view showing a cell culture processing system different from the embodiment of FIG. FIG. 3 is a partially cutaway perspective view of the transport device 11 of the present invention. FIG. 4 is a perspective view showing the transfer robot 22. FIG. 5 is a perspective view showing the opening / closing door 51. FIG. 6 is an explanatory diagram for explaining the communication operation of the opening 24. FIG. 7 is a perspective view showing the attaching / detaching means 16. FIG. 8 is an explanatory diagram for explaining the connecting operation of the attaching / detaching means 16. FIG. 9 is a control block diagram of the cell culture processing system. FIG. 10 is a perspective view showing a culture plate and the like.

  A cell culture treatment system consisting of a transfer device and a treatment device used for various types of culture processes has been realized with a minimum floor-occupying area and reduced processing time.

  FIG. 1 is a plan view of an embodiment of the cell culture treatment system of the present invention. 1a is a cell culture treatment system, 11 is a first treatment device (conveyance device), 12 is a dispensing device, 13 is a constant temperature device, 14 is a cell sheet laminating device, 15 Is a loading / unloading device such as a culture plate (second processing devices 12 to 15), and 16 is a detaching means. An attachment / detachment means 16 is provided on the side surface of the hexagonal conveyance device 11 in the center of the drawing. The attaching / detaching means 16 includes a cell culture device, a dispensing device 12, a thermostatic device 13, a cell sheet laminating device 14, and a carry-in / out device such as a culture plate (the devices shown in 12 to 15 are called processing devices). It is detachably connected. The transfer device and each processing device are provided with a sealed space (housing) that is shielded from the outside world, and becomes a single continuous closed space after the transfer device and each processing device are connected. Decontamination gas is supplied into each processing apparatus by connecting a decontamination / sterilization gas supply port through which each decontamination / sterilization gas can be introduced to each processing apparatus. . As a result, decontamination and sterilization are possible throughout the cell stacking system 1a.

In such a cell culture treatment system 1a, each treatment apparatus is covered with a casing, and a clean atmosphere (class 1000 or less) is provided by providing a fan filter unit (not shown) in which a HEPA filter and a fan are combined (not shown) at the top of the casing. ).

Since the cell culture treatment system 1a can be connected to the treatment apparatus via the attaching / detaching means 16, the number of apparatuses can be increased or decreased within the connectable quantity range. For this reason, the combination of each processing apparatus can be changed so that it can respond to culture | cultivation processes of many kinds of cells, tissues, and the like. There is an advantage that it can be used properly depending on the application. For example, storage of a cell suspension, cell seeding on a culture plate, medium exchange, stacking of cell sheets, shipping of stacked cell tissues, and the like.

A cell culture treatment system 1b shown in FIG. 2 is an embodiment different from that shown in FIG. This cell culture processing system 1b includes various processing devices connected to the outer periphery of the transport device 11a, and a delivery device 17 is provided on the left side of the transport device 11a via an attaching / detaching means 16, and the delivery device 17 is further attached to the left side. Another conveying device 11b is connected via the means 16. According to such a cell culture treatment system 1b, the number of treatment devices can be increased without being limited to the number of attachment / detachment means 16 that can be attached to one transport device. For this reason, it is possible to easily change the layout of the processing apparatus due to the progress or change of the culture process, and it is possible to easily increase the number of processing apparatuses accompanying the progress and complexity of the process and the increase in production volume. Here, the delivery device 17 includes, for example, a mounting table such as a culture plate and a decontamination gas supply port for supplying decontamination gas and disinfecting gas to the enclosure. An opening / closing door that can seal and open the opening for transferring the culture plate is provided. Thereby, a culture plate etc. can be conveyed without a foreign material mixing a culture plate etc.

Moreover, according to such a cell culture processing system 1, it is possible to combine a plurality of processing apparatuses having the same function, to operate the processing apparatuses alone in parallel, or to operate the processing apparatuses alone in parallel. This enables high-throughput and efficient production.

The cell culture treatment system 1 is configured by combining treatment devices according to a process such as culturing of cells and tissues. Each processing device is equipped with a command for operating an actuator such as a motor, and includes a control device for transmitting / receiving and recording signals from various sensors such as environmental monitors, electrical information such as measurement information and operation logs. In addition, the cell culture processing system 1 coordinates the operation between the processing apparatuses, and includes measurement results (operation log and environment log) including ID information and environmental information of the processing apparatus that has performed the culture processing process, and cells. An integrated control device is provided that stores and manages information associated with sample information such as tissue. By transmitting / receiving an electrical signal from the control device to the integrated control device in association with the ID information of the processing device, any processing device can be electrically connected even when one processing device is removed from the transport device and replaced with another processing device. The integrated control device can recognize whether signals have been exchanged. As a result, it is possible to deal with various culture processes, save labor, efficiently produce cells and tissues, and reduce manufacturing costs.

The integrated large-scale system equipped with all the functions necessary for culture processing is difficult to interrupt the processing of cells from one patient while processing cells from one patient, and the time occupied by the device is long. On the other hand, since the processing apparatus can be attached and detached as necessary, the apparatus occupation time can be easily adjusted. When a part of the processing apparatus breaks down, only that part can be separated and repaired, so that the influence on the entire system can be minimized. These features are effective in the medical field where irregular treatment is required for each patient and the treatment schedule is different, and a quick response to the problem is required.

It has a function to manage the culture processing procedure of cell tissues and has a communication means to transmit the measurement results of the culture progress and culture environment, instructing each processing device to execute the culture processing process, and organizing and storing the data Because it has a control device with the function to perform, it is possible to produce in accordance with GMP (Good Manufacturing Practice).

FIG. 3 is a partially cutaway perspective view of the first processing apparatus (conveyance apparatus 11) of the present invention.
21 is a base that is hexagonal when viewed in plan, 22 is a transfer robot provided in the upper center of the base 21, and 23 is a shield for the SCARA-type transfer robot 22 from the external atmosphere (of the casing). 24 is an opening for carrying in / out objects to be conveyed between the outside of the casing 23 and the inside of the casing, and 25a and 25b are openings. This is an open / close door that seals and releases 24. Reference numeral 16 denotes attachment / detachment means for attaching / detaching the transport device 11 attached to the side surface of the base and another dispensing device 12 or the like. For this reason, the attachment / detachment means 16 can be coupled to and detached from the transport device 11 while maintaining hermeticity and sterility. For this reason, safety is ensured without internal cells and tissues being contaminated by various germs, other cells, and the like. In this embodiment, in order to further keep the inside of the processing apparatus so that the amount of dust is minimized, an air cleaning device 28 using a hepa filter 27 or the like may be provided.

Further, the fan 29 at the upper part of the ceiling 23b of the transport device 11 is provided with a decontamination / sterilization gas supply pipe 35 (decontamination / sterilization gas supply port 36) from the decontamination / sterilization device, In addition, a decontamination / sterilization gas exhaust pipe 37 (decontamination / sterilization gas supply port 38) for exhausting and collecting decontamination / sterilization gas from the inside of the housing and sending it to the decontamination / sterilization device is provided. Decontamination and sterilization are performed before introducing the cells into the sealed space of the processing apparatus in order to kill germs and cells adhering to the inside of the sealed space of the processing apparatus and the outside of the introduced sealed container. Furthermore, dust in the sealed space is also removed by a filter such as a hepa filter. Various decontamination means such as hydrogen peroxide vapor and ozone are conceivable for the decontamination / sterilization equipment, but effective and safe are ensured, and decontamination / sterilization validation can be established. To do. This decontamination / sterilization is possible even when a single processing device or a plurality of devices are connected. In addition, when the processing devices are connected, they are exposed to the outside, and when connected, they are surrounded by a surface and sealed. Decontamination and disinfection are possible even in a small space.

The housing 23 has a slightly larger internal volume than the operating range of the SCARA type transfer robot 22. In the case of a SCARA type transfer robot 22 or an arm slider type transfer robot, the internal space shape of the housing is a cylindrical shape. In the case of an articulated type transfer robot, a hemispherical shape is used to minimize the internal volume. can do. In addition, a gas supply port and an exhaust port for supplying decontamination gas into the housing are provided at the top of the housing. As a result, by supplying a minimum amount of hydrogen peroxide gas to decontaminate the inside of the enclosure, it reaches a gas atmosphere of a predetermined concentration in a short time and maintains a predetermined decontamination / sterilization strength. The decontamination / sterilization process can be completed in a short time.

As the material inside the housing, a member that is less deteriorated by a decontamination agent such as hydrogen peroxide gas is used, and appropriate caulking is applied to the joint portion of the member to maintain a predetermined sealing degree. For example, the metal inside the casing is stainless steel, and the movable part and the caulking material are silicone rubber. When using a member that deteriorates over time due to hydrogen peroxide gas, the structure should be easy to replace. It is possible to provide a device that can be used for a long time with minimal deterioration due to gas.

Each processing device does not enter the room like a clean room, and because it is a minimum size space, it is easy to shorten the decontamination / sterilization time, maintain sterility, and inexpensively. Can be manufactured. Moreover, since combinations are possible as necessary, the entire system combining the first processing apparatus (conveying apparatus) and the second processing apparatus (each processing apparatus) is inexpensive. Therefore, the cost of cells and tissues to be produced can be reduced.

The SCARA-type transfer robot 22 shown in FIG. 4 is installed in the center of the upper part of the base 21 in a room covered with a housing 23. As a result, the distance from the turning center line 31 (indicated by the one-dot chain line) of the SCARA-type transfer robot 22 to the open / close door and the (shortest) distance R to the inner wall surface of the housing are constant.

Since the SCARA type transfer robot 22 can minimize the operating range, the internal volume of the housing 23 can be minimized. For this reason, when supplying gas for decontamination, it is possible to reach a predetermined decontamination / sterilization strength in a short time by supplying a minimum amount of gas. In addition, if the volume inside the housing 23 is large, gas does not flow inside the housing 23, and there are places where the structure becomes complicated and cannot be completely decontaminated. Can be prevented.

In the SCARA type transfer robot 22, a robot base 41 is attached to the base 21 via a flange 42. On the upper part of the robot base 41, an elevating means 43 for elevating the SCARA type transfer robot 22 is provided. The elevating / lowering means 43 makes the movable member (not shown) linearly movable in the Z direction in the drawing by rotating the ball screw shaft by the operation of the motor. In the upper part of the movable member of the lifting / lowering means 43, there is provided a turning means 45 for turning the SCARA-type transfer robot 22 in a direction indicated by θ in the figure in a horizontal plane. The turning means 45 includes a bearing member such as a bearing, a motor, and the like and a turning shaft member 45, and a turning center line 31 (indicated by a one-dot chain line in the drawing) of the turning shaft member is oriented in the vertical direction. A second arm 46 is provided at the end of the turning shaft member 45, and a second arm 47 having a rotation axis parallel to the turning center line 31 is provided above the second arm 46. An end effector having a rotation axis parallel to the turning center line 31 on the upper portion of the arm 47 and placing a microplate 48 serving as a transported object on the upper portion is provided. The SCARA transfer robot 22 is linearly driven by a second arm 46, a second arm 47, and an end effector 48 that are synchronized with each other by a pulley, a rubber belt, or a steel belt attached to the rotation shafts of the second arm 46 and the second arm 48. A motion or a desired curve motion can be performed. Note that the various actuators provided in the SCARA-type transfer robot 22 are operated based on electrical signals from the control device 50 (indicated by a thick frame in the figure). As a result, the SCARA-type transport robot 22 transports a microplate, a bottle, a disposable chip of a dispensing device (consumable supplies supplied to various devices) or the like inside the decontamination / sterilization atmosphere covered with the housing 23. Can do.

With such a SCARA-type transfer robot, the second arm 47 and the second arm 48 are rotated in a horizontal plane so as to transfer the object to be transferred. Therefore, the movable range in the height direction during the transfer operation is minimized. Can be suppressed.

An O-ring such as a fluorine-based resin or a seal member is attached to the rotating portion of the SCARA type transfer robot 22. This prevents the decontamination gas from flowing into the SCARA-type transfer robot 22 and corroding the internal members.

A resin bellows seal 44 is attached between the periphery of the robot base 41 of the SCARA-type transfer robot 22 and the periphery of the movable member of the elevating means 43. For this reason, even when the SCARA-type transfer robot 22 is moved up and down, gas for decontamination and sterilization is prevented from flowing into the SCARA-type transfer robot 22 and corroding the internal members.

  The end effector has a flat plate shape, on which a culture plate or the like can be placed. The outer periphery of the culture plate or the like placed above the end effector is provided with a conical protrusion 49 or a guide member for guiding the four corners or side surfaces of a substantially rectangular parallelepiped culture plate. Can do.

In addition to the SCARA-type transfer robot, an industrial articulated robot, a slider robot, or the like may be used as the transfer means.

  The open / close door 25 a shown in FIG. 5 includes a driving unit on a base, and a door member 56 is attached to a movable member 55 a of the driving unit 55. The door drive means 55 is an air cylinder, and can be opened and closed by moving up and down. One end of the bellows seal 57 is attached to the periphery of the drive means 55 on the base 21 and the other end is attached to the movable part. This prevents gas for decontamination and sterilization from flowing into the door driving means and corroding the internal members. FIG. 5A shows a state where the door member 56 is lowered by the operation of the door driving means 55 and the opening 24 is in communication with the outside of the housing 23. FIG. 5B similarly shows a state where the door member 56 is raised and the opening 24 is closed, and the inside of the housing 23 is sealed. A silicone seal 57 is attached to the door member 56, the housing 23 (around the opening 24), and the contact portion (on the door side or the housing side). This silicone seal is in the form of a tube, and it may be one that seals between the bulge and the door member 56 by injecting compressed gas into the tube. You may seal.

  The open / close door 25b shown in FIG. 3 is a revolving door having a rotation axis parallel to the wall surface 23a of the housing 23. A silicone seal is attached to the door member 56, the housing 23 (around the opening 24), and the contact portion (on the door side or the housing side). In such a door, it can be set as the structure which rotates a door by attaching a motor or a cylinder, and a link member to a rotating shaft. In such a door, sealing can be performed by crushing the silicone seal evenly between the door and the wall surface inside the housing by rotational force. Fluorine-based resin material O-rings or fluorine-based V seals are used at the rotational drive location, so that gas for decontamination and sterilization flows into the inside of the rotating means and the internal members corrode. Is preventing.

FIG. 6 is an explanatory diagram for explaining a state when the first processing apparatus (conveying apparatus) and the second processing apparatus are connected. 23a is the wall surface 23a of the housing 23 of the first processing apparatus. The right side of the opening / closing door 56 (the area indicated by A in the figure) is inside the casing, and the left side (the area indicated by C in the figure) is outside the casing of the first processing apparatus (conveyance apparatus) 11. Left side of the door member 56 of the second processing device (the region indicated by B in the figure is inside the housing 23c of the second processing device. The region indicated by D is outside the first processing device and the second processing device. However, when the openings 24 and 24a of the first processing apparatus and the second processing apparatus are communicated with each other, this is an area inside the housing.

FIG. 6A shows a state before the first processing device and the second processing device are connected. FIG. 6B shows a state in which the second processing apparatus (left side in the figure) is moved from the state of FIG. 6A to the right side in the figure until the position immediately before the seal member 58c and the seal receiver 58b come into contact with each other. ing. In FIG. 6C, the seal receiver 58b and the seal member 58c are brought into contact with each other by inflating the gas into the seal member 58c having an inner tube shape, and the periphery of the openings 24 and 24a from the outside. It shows a state of being shut off (sealed inside). The region indicated by D in the figure is a region isolated from the outside air and the inside of the housing in the state of FIG. At that time, decontamination / sterilization gas is supplied to the region D from the decontamination / sterilization gas supply port 35a to the region D (in some cases, a decontamination / sterilization gas exhaust recovery port (not shown) is provided) Area D is decontaminated and sterilized. In this way, when the opening communicates, foreign matter that adversely affects cell culture inside the housing can be prevented. In FIG. 6 (D), the gas is introduced from the sealing member 58a provided in the housing 23a of the first processing apparatus and inflated by being in contact with the door member 56 from the tube-like seal member 58a. The state which extracted and made the area | region A and the area | region D inside the housing 23a communicate was shown. At the same time, from the state where the gas flows in from the sealing member 58d which is provided in the housing 23c of the second processing apparatus and has an inside and is inflated to contact the door member 56a, the gas is removed and the housing 23c is removed. A state where the inner region B and the region D are communicated is shown. Accordingly, a gap is provided between the seal member 58 and the door member 56, and the seal can be prevented from being worn by the opening / closing operation of the opening / closing door.

  The attaching / detaching means 26 shown in FIG. 7 is provided at the lower part of the side surface of the base 21, and includes a positioning pin and 61, an L-shaped key member 62 provided so as to protrude perpendicularly to the side surface, and a horizontal straight line. A key driving means for moving forward and backward and rotating in a direction, and an attachment / detachment detection sensor 64 for detecting that the conveying device and the dispensing device are connected or separated by the operation of the attaching / detaching means 26 are provided. Reference numeral 65 denotes a connector, a connector that transmits and receives electrical signals between the integrated control device and the processing device, a connector that supplies power to the second processing device, and a compressed gas, vacuum, or other gas (carbon dioxide, Nitrogen) A connector such as a connector for supplying a liquid. Reference numeral 66 denotes a pipe connector for supplying decontamination gas from the decontamination / sterilization apparatus via the pipe. Reference numeral 67 denotes a pipe connector for collecting exhaust of the decontamination / sterilization gas exhausted from the inside of the housing of the processing apparatus, and the decontamination / sterilization gas exhaust recovery port is piped to the decontamination / sterilization apparatus.

  In this embodiment, the attaching / detaching means 26 operates the key driving means 26a to advance toward the device for connecting the key member (in the arrow direction shown in FIG. 7A), and then moves the key member (the rotation direction shown in FIG. 7B). (2) After rotating, the key member is moved backward (in the direction of the arrow shown in FIG. 7C). At this time, the positioning pin 61 that protrudes from the wall surface of the base is attached to the side surface of the base of the second processing device (dispensing device or the like) to be connected and is attached to a position corresponding to the positioning pin mounting position. It fits with a guide member (a member having a recess and a hole that is slightly larger than the outer shape of the positioning pin). Thereby, a conveyance apparatus can draw in a dispensing apparatus and can connect with a predetermined position. A sensor 64 that protrudes from the wall surface detects when the processing device is connected, and transmits an electrical signal to the control device or the integrated management processing device that the transport device and the processing device are connected. In addition, this connection also allows communication line connectors to communicate (in the case of a wireless LAN connection, receiving an electrical signal from a sensor, etc.) ID information of the processing device itself connected from the control device of the second processing device. By sending an electrical signal to the integrated control device, the integrated control device can recognize which second processing device is connected to the first processing device.

  By detecting that the pin has been inserted by the sensor and starting the pull-in operation, it is possible to provide a configuration that supports the attachment / detachment. The pull-in operation will be described with reference to FIG. FIG. 8A shows a state where the attaching / detaching means 16 of the first processing apparatus and the attaching / detaching means receiver 16a of the second processing apparatus are separated and not connected. FIG. 8B shows a state where the sensor 64 and the sensor receiver 64a are in contact with each other when the second processing apparatus is moved to the right side in the drawing. The distal end of the positioning pin 61 is inserted into the recess of the positioning pin receiver 61a. Thereby, a 1st processing apparatus and a 2nd processing apparatus can be smoothly connected, without raise | generating a position shift at the time of attachment or detachment operation. An electrical signal for starting the connection operation from the sensor 64 is sent to the control device 50. Thereby, the operation of moving the key member 62 toward the attaching / detaching means receiver 16a is performed (the state shown in FIG. 8B). Thereafter, the key member rotates by the operation of the key driving means 62a of the key member 62, and engages with the key member receiver 62a (the state shown in FIG. 8C). Thereafter, the key member is retracted by the operation of the key driving means, so that the attaching / detaching means 16 can draw the attaching / detaching means receiver 16a toward the first processing apparatus, and the first processing is performed so that the second processing apparatus does not move. Can be fixed to the device.

Thereby, the combination of each apparatus processing apparatus can be changed by increasing / decreasing a processing apparatus via an attachment / detachment means with respect to a conveyance apparatus, and can respond to culture | cultivation processes of many kinds of cells, tissues, etc.

Time required for the process because it is possible to combine a plurality of processing devices with the same function provided with attachment / detachment means on the transfer device, or to operate the processing devices alone in parallel or at the same time. By absorbing the difference, efficient production is possible.

The attaching / detaching means 16 is provided between the processing apparatus and the base 21 of the cell culture apparatus 11, and the mechanism can exert its function regardless of which side it is provided. For convenience of device design, the processing device and the transport device (for example, the base of the processing device or the base of the transport device) may be provided on either side. As another embodiment of the attaching / detaching means 16, for example, a block member having a recess on the upper surface of the member is provided on the floor, and positioning can be performed by inserting a pin from above, so that the connector can be connected and disconnected by attaching / detaching operation. It may be an attaching / detaching means attached with the connector facing in the direction.

The transport device 11 of the present embodiment is connected to the control device via a wiring, and can operate various actuators in response to an electrical signal from the control device, thereby removing decontamination / sterilization gas. The actuator or the decontamination / sterilization apparatus can be operated by receiving an electric signal from the control device directly or indirectly because it is connected to the decontamination / sterilization apparatus via a pipe. Further, since it is connected to the compressed gas or the vacuum pump device via a pipe, the actuator or the compression pump device and the vacuum pump device can be operated by receiving an electrical signal from the control device directly or indirectly. In addition, power is supplied because it is connected to the power supply means via wiring. In this embodiment, a connector for wiring that transmits and receives electrical signals from the control device, a connector for piping that supplies and exhausts decontamination / sterilization gas from the decontamination / sterilization device, and a power source It is good also as a structure provided with a connector. By arranging and attaching a connector at a position corresponding to each connector of the second processing apparatus (dispensing apparatus or the like) to be connected, an electrical signal or the like from the control apparatus can be transmitted and received by connection.

The second processing device (dispensing device or the like) may include a vacuum pump, a compressor, a battery, an electric signal receiver using a wireless LAN, and a processing device control device in addition to the transport device. By connecting to the housing of the transport device, power may be turned on to the processing device control device, or the processing device may be activated (made available) by transmitting / receiving a communication start signal to / from the integrated control device. .

Processing equipment includes transport equipment, carry-in / out equipment, isolated primary culture equipment, subculture equipment, thermostatic equipment, tube connection robot, automatic warehouse equipment, transport lane equipment, and other culture, dispensing, transport, and measurement (weighing).・ Analytical (observation) devices and storage devices for goods (for example, microplates, tube racks, disposable chips, bottles, reagent tubes and other accessories), measuring devices, loading and unloading devices such as plates, and decontamination / sterilization Refers to devices.
In addition to the above treatment devices, various treatment devices are used depending on the cells, tissues, and organs to be cultured. For example, 3D organizing equipment that forms 3D new tissue using 3D scaffolds, various bioreactors and devices that give stress, strain such as pressure, tension, compression, etc., packaging for packaging Including devices.

The first processing device (conveying device) is a base, a robot that transports culture plates and the like provided on the base, and the robot's movable atmosphere is shielded from the outside air (covers the side and ceiling of the device and decontaminates it).・ Prevents sterilization from filling the inside of the gas and preventing it from leaking to the outside), an open / close door that opens and closes the opening for transferring culture plates etc. inside and outside the body, and supplying decontamination gas to the inside of the body It can be set as the structure provided with a decontamination apparatus and attachment / detachment means. Moreover, it can be set as the structure which equips the lower part of a 1st processing apparatus with a moving means as another structure.

Second treatment device (carrying in / out device, cell isolation primary culture device, subculture device, constant temperature device, tube connection robot, automatic warehouse device, transport lane device, cell seeding device, medium dispensing device, cell sheet stacking Equipment, other culture / dispensing / transport / measurement (metering) / analysis (observation) equipment, storage equipment for goods (for example, microplates, tube racks, disposable chips, bottles, reagent tubes and other accessories), measurement equipment , Plate loading and unloading equipment, and decontamination equipment, etc. are provided on the base and the upper part of the base so that the side and ceiling of the equipment are covered. A housing that prevents leakage to the outside, an opening for transferring culture plates and the like inside and outside the housing, an open / close door that can open and close the opening, and movement of a roller or a car at the bottom of the base With means Dispensing device, loading and unloading device, isolated primary culture system, subculture device, thermostat, tube connecting robot, automatic storage device, a transport lane device.

The second processing apparatus will be described in detail below.
The carry-in / out device includes a door for carrying in equipment, and cells, culture solution, medicine, culture dish, centrifuge tube, and other equipment necessary for culture are introduced into the carry-in / out apparatus from this door.
The loading / unloading device is equipped with an operation glove made of an elastomer that enables workers to move, open, and open the equipment from the outside of the loading / unloading device. It can move to the vicinity. Instead of the operator, a belt conveyor or a robot may be provided inside and used for movement.

<Isolated primary culture device>
The isolated primary culture device includes a fragmentation device for subdividing the introduced tissue piece from the carry-in / out device by the transfer robot of the culture treatment device. What is introduced by the transfer robot is a tissue tube, a centrifuge tube, a petri dish, a culture solution, and the like. Although the means of subdivision is not limited, a method of minimizing damage to cells is taken. Moreover, the cell separation apparatus for taking out a cell is provided. The cell separation apparatus decomposes the extracellular tissue of the fragmented tissue pieces with an enzyme, rotates (while decomposing), and isolates the cells. Furthermore, the target cell can be taken out by utilizing the difference in cell adhesion, cell size and the like.

  The isolated primary culture apparatus includes a mixer as a subdividing apparatus for subdividing the cell debris, and a moving means 8 (for example, a separator) that puts the cell debris from a container (for example, a bottle or a plate) containing the cell debris into the mixer. Note means) can be provided.

Moreover, the primary culture apparatus for increasing the taken-out cell further is provided. The primary culture apparatus is equipped with a thermostat capable of maintaining an appropriate gas concentration, where cells are grown in a culture solution until the number of cells that can be subcultured in an appropriate environment is reached.

The isolated primary culture apparatus is equipped with a medium supply means for supplying the medium to the fragmented cell pieces that take out the fragmented cell pieces from the mixer, and a constant temperature shaking means for shaking the cells while maintaining the cells at 37 ° C. It can be set as the structure provided.

  In this example, the tissue subdivision device, the cell separation device, and the primary culture removed from the living body are combined to form an isolated primary culture treatment device. However, the subdivision treatment device, the cell separation treatment device, the primary culture device, The culture treatment device and each may be a separate treatment device.

<Subculture device>
The subculture apparatus is an apparatus that further increases the number of cells after the primary culture to proliferate to the number of cells sufficient for treatment.
In this example, the cells are cultured in a closed closed container. By culturing a large amount of cells, it becomes a large device, so it is not directly connected to the culture treatment device, but the cell suspension made up of the closed closed container and the isolated primary culture treatment device is connected to the tube. Then, it is transported to a closed closed container in the primary culture treatment apparatus. Of course, the processing apparatuses may be coupled to each other and transferred by a transfer robot. If necessary, a plurality of the same treatment apparatuses may be connected and cultured.
The culture medium can be proliferated by periodically exchanging the culture solution in an appropriate environment in the thermostatic apparatus provided in the subculture apparatus.

<Dispensing device>
The dispensing device uniformly and accurately seeds a predetermined number of temperature-suspended cell suspensions on a predetermined number of temperature-responsive culture dishes, removes the culture medium from the culture dish as necessary, and supplies a predetermined amount of fresh culture medium. Is a device for adding.
The cell suspension transported by the tube from the subculture apparatus is stored in the lamination processing apparatus and is then seeded on a plurality of temperature-responsive culture dishes, or directly seeded on the plurality of dishes. Cells in the seeded suspension adhere uniformly to the temperature-responsive culture dish.

<Cell sheet stacking device>
The cell sheet laminating apparatus is an apparatus for superposing a plurality of sheet-shaped cell sheets to create a transplanted neoplastic tissue having a very high cell density.
The thin sheet-like cells formed on the cell-adhesive surface of the temperature-responsive culture dish are separated from the dish by changing (decreasing) the temperature. Is taken out and a plurality of sheets are overlapped by a robot or an actuator. The stacked cell sheets are integrated into a new tissue with a very high cell density.
In this way, the cell sheet laminating apparatus is responsible for the step of laminating a plurality of cell sheets through cell seeding, cell sheeting, and cell sheet peeling to create a transplantable new tissue.

<Constant temperature device>
The thermostatic device (incubator) is used to appropriately maintain gas concentration, temperature, and humidity for growing cells and storing new tissues until transplantation.

<Tube connection robot>
In order to move the cell suspension or the like in the closed type culture section to the closed type or aseptically sealed space of another processing apparatus, to connect or disconnect the tube to or from the destination processing apparatus. Maintains sterility and airtightness even when the socket (connector or connector) is connected or disconnected. Such an apparatus can be used when a suspension is transported between processing apparatuses by a tube.

<Automatic warehouse equipment>
The automatic warehouse device stores the culture fluid (medium), growth factors, drugs, cells, containers, etc. necessary for the cells and tissues to be cultured, and automatically unloads them according to the cells and tissues to be cultured. It supplies to each processing apparatus from the apparatus carrying-in-like door part of an apparatus and each processing apparatus.
When automatic supply is not performed, a pass box is installed to carry each equipment in place of the automatic warehouse.

<Transport lane device>
The transfer lane device serves as a guide for guiding to a predetermined place in order to connect the sealed space of each processing device and the tube of the tube connecting device, and a metallic tape, rail, guide post, or the like is used. Is called. In addition, such an apparatus can be used when connecting a tube automatically.

<Other devices>
In addition to the above processing devices, various processing devices are used depending on the cells, tissues, and organs to be cultured. For example, 3D processing equipment that forms 3D new tissue using 3D scaffolds, various bioreactors and devices that give stress, strain such as pressure, tension, and compression, and packaging for transportation A packaging device may be added.

  In the place where the outside of the opening 24 of the housing 23 of the present embodiment and the outside of the opening of each processing apparatus (dispensing device, etc.) to be connected come into contact with each other, the inside of the opening is sealed to both or either one. A silicone seal member is attached. As a result, the internal sealability can be maintained even after the devices are connected to each other, and the gas does not leak from the connection portion even when the decontamination gas is supplied into the enclosure.

<Integrated control device>
The transport apparatus according to the present embodiment includes a control device that transmits and receives electrical signals to and from the transport apparatus. The control device can transmit and receive electrical signals to and from the integrated control device.
The integrated control device coordinates the operation of each processing device (operates by sending and receiving electrical commands to individual processing devices according to the pre-gram), or records the progress of culture and environmental changes, and stores the database Manage (collect and manage information from the control device of each processing device). Further, the integrated control device outputs the data to an external printer, computer, or storage device in response to a user request.

The integrated control device includes a communication means for sharing data and exchanging signals with each processing device. Even if the communication means is RS23C iEEE, or wired or wireless, it is sufficient to solve the security problem. Furthermore, an input device is provided for inputting conditions for culturing, inputting cell or patient IDs, etc., and giving instructions for starting operation, culturing status, browsing the database, and the like. Furthermore, a display device for displaying culture condition settings, cell or patient ID, culture status, and the like is provided.

The integrated control device may be separated into a control device that coordinates the operation of each processing device, and a database management device that records the progress of culture and changes in environmental conditions and manages the database.

FIG. 9 is a control block diagram of the cell culture processing system.
Each processing device may be provided with an input device or a display device so that it can be used independently.

The cell culture processing system 1a includes an integrated control device 70a and a data management device 70b. Reference numeral 50 denotes a control device of the transport device 11. 71a is a control device for the dispensing device 12, 71b is a control device for the thermostatic device 13, 71c is a control device for the cell sheet laminating device 14, and 71d is a loading / unloading device such as a culture plate. 15 control devices.
The integrated control device 70a includes an input device for setting culture conditions, inputting information on cells to be cultured, and instructing to start a culture process, and a display device for displaying culture conditions, cell information, operating conditions, and the like. In addition to receiving information from each processing device, it transmits an instruction signal and includes a communication means for sending data sent from each processing device to the data management device and a control device for controlling each processing device.
Each processing apparatus can be operated independently, but is determined in advance by an integrated control apparatus 70 that performs communication between the processing apparatuses, executes a culture treatment process, and performs a fully automatic operation that links the processing apparatuses. Automatic operation is performed. Each processing apparatus and the integrated control apparatus 70 communicate with each other by a run cable, a wireless run, or other communication means. Data such as progress information of the operation process, culture environment data, and pass / fail judgment of the culture process is sent from each processing apparatus to the integrated control apparatus 70. From the integrated control device, each processing device is instructed to start a decontamination / sterilization process or a culture process of each processing device.

The data management device 70b is an input device for inputting instructions such as data retrieval and storage destinations, a database management device for organizing sent data and creating a database, a display device for displaying instruction contents and results, An output device for outputting data to a storage device or a printer is provided.

The sealed space of each processing device decontaminated and sterilized by the sterilization / sterilization device is combined as necessary, and the combined sealed space maintains sterility and is removed from the living body in the sealed space. The tissue introduced into is subdivided and degraded with enzymes, and the cells are separated and isolated. Thereafter, primary culture, amplification culture, and organization are performed to obtain a transplantable cell suspension or a new tissue that is maintained in a sterile state. Cell suspensions and new tissue are inspected, packaged and shipped, and various data in the culture process are preserved.

The example which has shown the role assignment of the control apparatus of the processing apparatus of each process, and an integrated control apparatus is shown. The division of roles from A1 to A16 is as follows.
A1. First, the input device of the integrated control device manages the use authority of the operator with an ID and a password, and distinguishes possible operation contents according to the operator's authority. Necessary items such as patient ID and culture conditions are input from this input device.
Thereby, in the control apparatus in the integrated control apparatus, an appropriate operation procedure program is selected from the programs of the various operation procedures incorporated.
A2. After the input is completed, a signal indicating completion of operation preparation of the processing device used for operation is transmitted from each processing device to the integrated control device.
A3. The integrated control device confirms the operation completion signal from each processing device involved in the operation. A4. When the confirmation is completed, the integrated control device issues a connection command for the processing device to each processing device.
A5. The connected processing device checks that the connection has been made with a sensor or the like.
A6. When it is confirmed that the connection has been completed reliably, a message is transmitted from each processing device to the integrated control device.
A7. The integrated control device issues an instruction to start the decontamination process to the processing device and the decontamination device.
A8. Decontamination is performed on the processing apparatus to which the decontamination / sterilization apparatus is connected.
A9. When the decontamination is completed, a decontamination completion signal is transmitted from the decontamination device and the decontaminated processing device to the integrated control device. (If there is a processing unit that operates independently, decontaminate that processing unit as well. The decontamination status of each processing unit (decontaminated or not decontaminated) is displayed in the integrated control unit along with the processing unit ID. Remembered.
A7. The integrated control device that has received the signal indicating completion of decontamination / sterilization gives an operation command based on the optimal program for the processing device to be operated first.
A8. The processing device that has received the operation command operates with the instructed program.
A9. The processing apparatus in operation regularly sends data of predetermined items such as operation time, environmental data, and checkpoint confirmation results to the integrated control apparatus. These data may be stored in each processing apparatus.
A10. The integrated control apparatus stores the sent data in a database.
A11. When it is detected that a malfunction or abnormality has occurred during operation of the processing apparatus, this data is transmitted to the integrated control apparatus. Depending on the contents, the restoration work is automatically performed inside the processing apparatus. When recovered, the data (contents, time, etc.) is transmitted to the integrated control device.
A12. The integrated control device stores abnormal data and repair data in a database.
A13. When the program in the processing device is completed, an end signal is transmitted to the integrated control device.
A14. The integrated control device transmits a separation command when it is necessary to separate the processing devices, and transmits a separation command when it is necessary to connect the processing devices. When binding in the middle, it is performed after confirming the decontamination / sterilization state from the device ID of the processing device to be newly bonded.
A15. Thereafter, the operation of each of the following processing apparatuses is repeated from A2 to A14.
A16. When all the processes are completed, various data sent from each processing apparatus are stored as a series of data for each sample, and are output to other storage devices or printers as necessary.

The culture plate and the like include the culture plate shown in FIG. 10A, the reagent bottle 81, the reagent bottle bracket 82, and the sheet cell holding member plate of the cell sheet laminating apparatus shown in FIG. 10B.
A guide member 82a is attached to the reagent bottle bracket 82 so that the reagent bottle 81 can be conveyed in an upright state in the reagent bottle bracket 82 for conveying the reagent bottle 81.

  The decontamination / sterilization apparatus of the present embodiment includes a liquid supply unit that supplies a hydrogen peroxide solution, a vaporization unit that vaporizes the hydrogen peroxide solution, a gas supply unit that supplies a gas to each device, and each device. A gas pipe connecting between the gas supply means, a pipe for collecting exhaust from each device, and an exhaust pipe for exhausting the collected gas after detoxification through the catalyst.

  The hydrogen peroxide gas generated by the decontamination / sterilization device flows into the inside of each device housing, so that it exists inside the housing 23 (including actuators, sensors, exteriors of containers, etc. provided in devices such as transport devices). You can kill the germs. The decontamination device may be a device that generates ozone or the like instead of hydrogen peroxide gas.

This system can be installed and used in existing limited spaces such as hospitals, and it is possible to prepare cells and tissues for GMP compatible treatment even in such locations. it can.
It also contributes to the creation of a cell and tissue culture business that installs multiple systems and undertakes and cultures the tissues brought in from each hospital.

DESCRIPTION OF SYMBOLS 1 Cell culture processing system 11 Conveyance apparatus 12 Dispensing apparatus 13 Constant temperature apparatus 14 Cell sheet laminating apparatus 15 Carrying-in / out apparatus 16, such as a culture plate, Attaching / detaching means

Claims (5)

A cell culture treatment system in which a plurality of treatment devices including a first treatment device and a second treatment device are detachably connected to each other via attachment / detachment means,
The first processing device includes:
The base,
A robot for transporting a culture plate or the like provided on the base;
A housing that shields the movable atmosphere of the robot from outside air;
An open / close door that opens and closes an opening for delivering a culture plate and the like inside and outside the housing;
A decontamination device for supplying decontamination gas;
Attaching and detaching means for connecting to the second processing apparatus,
The cell processing system according to claim 2, wherein the second processing apparatus includes a moving unit in a lower part.   When the second processing apparatus is connected to the housing of the first processing apparatus by the attaching / detaching means, the decontamination apparatus of the first processing apparatus is connected to the second processing apparatus via the attaching / detaching means. 2. The cell culture treatment system according to claim 1, wherein piping for supplying a decontamination gas is connected.   2. The plurality of attachment / detachment means are provided, and the plurality of second processing apparatuses can be selectively attached to / detached from the housing of the first processing apparatus via the attachment / detachment means. Cell culture treatment system.   The cell culture treatment system according to claim 3, comprising a plurality of the first treatment devices. The cell culture treatment system according to claim 2, wherein the outside of the door is decontaminated by providing a gas supply port from the decontamination device outside the door.

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