JP6524784B2 - Stirring apparatus for cell culture system, and stirring method for cell culture system - Google Patents

Description

  The present invention relates to a cell culture system, and more particularly to a stirring device of the cell culture system and a stirring method of the cell culture system.

In recent years, in the fields of drug production, gene therapy, regenerative medicine, immunotherapy and the like, efficient mass culture of cells and tissues under an artificial environment is required.
In such cell culture, a culture vessel for use in cell culture and a culture vessel for supplying a culture medium to the culture vessel are connected by a tube to constitute a cell culture unit, and the cell culture unit is used to close the cell culture unit. By performing cell culture in a system, cells are proliferated while avoiding the risk of contamination.

In this cell culture unit, the medium container is generally placed in a cold storage room because the medium needs to be stored under refrigeration. In addition, since the cell culture requires control of temperature, air environment, etc., the culture vessel is placed in an incubator. Then, the culture medium is transferred from the culture medium container to the culture container via the tube.
Also, for example, Patent Document 1 discloses a culture apparatus that stores cells of this type in multiple stages and performs cell culture.

WO 2007/052716 pamphlet

Japanese Patent Application No. 2014-189273

  However, in the culture apparatus described in Patent Document 1, the medium container (medium bag) and the culture container (culture bag) are stored in the same thermostatic bath, and under the temperature environment suitable for each of these containers. Cell culture could not be performed.

  Therefore, the present inventors store a plurality of cell culture units including a culture container for carrying out cell culture by containing the medium together with the cells, and a culture medium container for supplying the culture medium to the culture container in multiple stages, We have developed a cell culture system (cell culture device) that can perform cell culture efficiently in a temperature environment suitable for each of the culture container and the culture medium container when performing cell culture individually for each culture unit (Patent Document 2) reference).

  In such a cell culture system, when the culture vessel is provided with a stirring device, the stirring device does not disturb the attachment and removal of the culture vessel to the cell culture system, and there is a risk of damaging the culture vessel. It is desirable to be small.

  The present invention has been made in view of the above circumstances, and in a cell culture system for culturing cells using a culture vessel, agitation that does not prevent attachment and removal of the culture vessel and has a low risk of damaging the culture vessel It aims at provision of an apparatus and the stirring method.

In order to solve the above problems, the stirring device of the cell culture system of the present invention is a stirring device of a cell culture system for performing cell culture using a culture container containing a medium together with cells, and the culture container is placed A loading platform, a base supporting the loading platform, an elongated stirring member movable horizontally while pressing the upper surface of the culture vessel, and movably supporting both ends of the stirring member; Two rails, which are horizontally provided on both the left and right opposite sides of the platform, which are longer than the width of the loading platform in the front-rear direction and movable vertically, and the respective rails and the base rotate The two sets of arms are pivoted about the connecting portion of the two sets of arms and the two sets of arms and the base, which are coupled so as to move the rails vertically while keeping the horizontal state. The first drive means to The stirring member on said two rails, and a second driving means for moving in the longitudinal direction, the rail upper surface, in a state that is disposed on the mounting base upper surface substantially coplanar, one end of the rail It extends to the outside of the width of the loading stand in the front-rear direction, and the stirring member is disposed outside the loading stand.

  The stirring method of the cell culture system of the present invention is the stirring method of the cell culture system by the stirring device of the cell culture system, wherein (a) the first driving means is the stirring member is the loading table. The arm is rotated until the stirring member is disposed at a first position which can horizontally move the upper side of the culture vessel without pressing the culture vessel from the state of being disposed outside, thereby the rail Is moved upward, (b) the second driving means moves the stirring member to the stirring start position on the rail, and (c) the first driving means is the first stirring member. The arm is rotated to move the rail downward until the stirring member is disposed at a second position where the upper surface of the culture vessel can be horizontally moved while pressing the culture vessel from the position of (D) said second drive Means for moving the stirring member to the stirring end position on the rail, and (e) the first driving means is arranged to position the stirring member from the second position to the first position The arm is rotated to move the rail upward, and the operations (b) to (e) are continuously repeated.

  According to the present invention, in a cell culture system for culturing cells using a culture vessel, it is possible to provide a stirring device and a stirring method with a low risk of damaging the culture vessel without preventing the attachment and removal of the culture vessel. Become.

It is a schematic plan view of the stirring apparatus of the cell culture system which concerns on 1st embodiment of this invention. It is a schematic side view of the stirring apparatus of the cell culture system which concerns on 1st embodiment of this invention. It is explanatory drawing which shows the operation | movement from the side view of the stirring apparatus of the cell culture system which concerns on 1st embodiment of this invention. It is a schematic plan view of the stirring apparatus of the cell culture system which concerns on 2nd embodiment of this invention. It is explanatory drawing which shows the clamp in the stirring apparatus of the cell culture system which concerns on 2nd embodiment of this invention, and its use condition. BRIEF DESCRIPTION OF THE DRAWINGS It is the appearance schematic of the cell culture system in each embodiment of this invention. It is an explanatory view showing an example of a cell culture unit stored in a cell culture system in each embodiment of the present invention. It is an explanatory view which carries out sectional view inside of an incubation tank of a cell culture system in each embodiment of the present invention. It is a schematic sectional drawing of the culture container storage shelf of the cell culture system in each embodiment of this invention.

Hereinafter, preferred embodiments of a stirring apparatus for a cell culture system and a stirring method for a cell culture system of the present invention will be described with reference to the drawings.
First, a first embodiment of a stirring device of a cell culture system and a stirring method of a cell culture system of the present invention will be described with reference to FIGS. 1 to 3.

First Embodiment
The stirring device of the cell culture system of the present embodiment is a stirring device of a cell culture system that performs cell culture using a culture container that contains a medium together with cells, and includes a loading table on which the culture container is placed and a loading table. A base to support, an elongated stirring member movable in the horizontal direction while pressing the upper surface of the culture vessel, and both ends of the stirring member are movably supported, and on both sides of the loading stand opposite to each other, the loading stand The two rails, which are horizontally longer than the width of the front and rear direction and movable vertically, are rotatably connected to the respective rails and the base so that the rails are maintained in the horizontal state. The first drive means for pivoting the two sets of arms about the connection between the two sets of arms and the base, and the stirring member on the two rails, And second driving means for moving in the front-rear direction, Lumpur upper surface, while being disposed in the mounting base upper surface substantially coplanar, stirring member, characterized in that arranged outside the stacking table.

Specifically, as shown in FIG. 1 and FIG. 2, the stirring device 10 of the cell culture system of the present embodiment includes the loading stand 11, the base 12, the rails 13, and the bending arms 14 (14-1 and 14-2). ), A stirring roller 15, and an endless belt device 16.
The loading table 11 is a flat table on which a culture vessel (culture bag) is placed. At four corners of the loading table 11, fixtures for fixing the culture vessel are provided upright. Holes are provided at four corners of the culture vessel, and the fixing tool of the loading table 11 is inserted into these holes, and the culture vessel is disposed on the loading table 11 in a planar manner.

The loading platform 11 is supported by a base 12 located below the loading platform 11 and fixed at a fixed height.
Two rails 13 which are longer than the width in the front-rear direction of the loading table 11 and movable in the vertical direction are horizontally provided on the opposite left and right sides of the loading table 11.

One end of a bending arm 14 is rotatably connected to the rail 13 on each of the front side and the rear side. The other end of the bending arm 14 is rotatably connected to the base 12. Then, by rotating the bending arm 14, it is possible to move the rail 13 in the vertical direction while maintaining the horizontal state.
In the initial state, the front end portion of the rail 13 is substantially at the same position as the front end of the loading table 11, and the rear end portion of the rail 13 is loaded by a length capable of disposing the agitating roller 15 on the rear outside of the loading table 11. It is located behind the rear end of the table 11.

  According to the stirring device of the cell culture system of the present embodiment, the height of the rail 13 can be changed by the rotation of the bending arm 14 as described above, so that the cost can be reduced without using an expensive slide rail or the like. It is possible to realize a stirring device with a simple structure.

  In FIG. 2, a bending arm 14-1 is a main moving portion, and a connecting portion 12-1 of the bending arm 14-1 and the base 12 is a rotation axis. Further, the bending arm 14-2 is a driven portion, and the connecting portion 12-2 of the bending arm 14-2 and the base 12 is used as a rotation axis. And the connection part 12-1 is rotated by the motor (1st drive device) which is not shown in figure, and the rail 13 is moved to an up-down direction. At this time, for example, the left connecting portion 12-1 in FIG. 1 is rotated by a motor, and this rotational movement is transmitted to the right connecting portion 12-1 in FIG. 1 to synchronize the operation of the left and right bending arms 14. Thus, the rail 13 can be moved in the vertical direction.

As the bending arm 14 in the present embodiment, an L-shaped arm, a V-shaped arm or the like can be suitably used. In the present embodiment, a linear arm may be used instead of the bending arm 14.
Moreover, in this embodiment, although the two left and right arms which each consist of two bending arms are used, it is not limited to this, You may use more bending arms.

  The stirring roller 15 is an elongated member (stirring member) which can move in the horizontal direction while pressing the upper surface of the culture vessel. At both ends of the agitating roller 15, wheels moving on the rail 13 are provided. In FIGS. 1 and 2, two wheels are respectively provided at both ends of the agitating roller 15, but the invention is not limited thereto.

  The stirring roller 15 is moved on the rail 13 by the endless belt device 16 (second drive device). The endless belt device 16 is fixed to the rail 13, has substantially the same length as the rail 13, and moves in the vertical direction as the rail 13 moves up and down. Both ends of the agitating roller 15 are fixed to a part of the endless belt in the endless belt device 16, and the wheels at both ends of the agitating roller 15 move on the rails 13 as the endless belt rotates. By this, the stirring roller 15 moves horizontally forward and backward on the culture vessel.

  At this time, for example, the rotation shaft of the endless belt in the endless belt device 16 on the left in FIG. 1 is rotated by a motor, and this rotational motion is transmitted to the endless belt device 16 on the right in FIG. In synchronization with the operation of the device 16, the stirring roller 15 can be moved on the culture vessel.

In the initial state of the stirring device of the cell culture system, the upper surface of the rail 13 is disposed substantially flush with the upper surface of the loading table 11. Further, the upper surface of the rail 13 may be disposed below the upper surface of the loading table 11. The substantially same plane also includes the case where the upper surface of the rail 13 is disposed slightly above the upper surface of the loading table 11.
At this time, the stirring roller 15 is disposed at the rear end of the rail 13 and is disposed at the rear outside of the loading table 11.

As described above, according to the stirring device of the cell culture system of the present embodiment, since the stirring roller 15 is disposed on the rear outside of the loading table 11 in the initial state, it is not present on the loading table 11. Further, the upper surface of the rail 13 is disposed on substantially the same plane as the upper surface of the loading table 11.
For this reason, when placing the culture vessel on the loading table 11, it is not necessary to make the culture vessel lie below the stirring roller 15, and it is possible to suppress the trouble of attaching and detaching the culture vessel to the cell culture system. Is possible.
In addition, since the rails 13 do not protrude from the surface of the loading table 11, it is possible to reduce the risk of damaging the culture vessel.

Next, the operation of the stirring device of the cell culture system of the present embodiment will be described.
In the stirring method of the cell culture system by the stirring device of the cell culture system of the present embodiment, (a) the first driving means presses the culture vessel from the state where the stirring member is disposed outside the loading platform The arm is rotated to move the rail upward until it is disposed at the first position where it moves horizontally above the culture vessel without moving the culture vessel, and (b) the second driving means moves the stirring member on the rail (C) the second driving unit moves the upper surface of the culture vessel in the horizontal direction while the stirring member presses the culture vessel from the first position. (D) the second driving means moves the stirring member to the stirring end position on the rail, and (e) the first driving means , The stirring member is disposed at the first position from the second position Uni arm is rotated to move the rail upward, subsequently, and repeating the operation of (b) ~ (e).

  Specifically, first, in the initial state, as shown in FIG. 3 (1), the stirring roller 15 is disposed at the rear end of the rail 13 and is disposed at the rear outside of the loading table 11. Further, the upper surface of the rail 13 is disposed on substantially the same plane as the upper surface of the loading table 11.

  Next, as shown in FIG. 3 (2), the bending arm 14 is rotated by the motor, and the stirring roller 15 (and the rail 13 and the endless belt device 16) is moved upward to the first position. Be placed. In the first position, the lower end of the stirring roller 15 is positioned above the upper surface of the culture vessel placed on the loading table 11. Further, the lower end of the agitating roller 15 is positioned above the upper end of the fixing tool for fixing the culture vessel placed on the loading table 11. By this, in the first position, the stirring roller 15 moves horizontally above the culture vessel without pressing the culture vessel and without being hindered by the fixtures provided on the loading table 11.

  Next, as shown in FIG. 3 (3), the endless belt device 16 moves the stirring roller 15 to the stirring start position (movement start position) on the rail 13. The stirring start position is a position at which the stirring roller 15 starts moving on the rear end of the culture vessel when the stirring roller 15 is moved downward.

  Next, as shown in FIG. 3 (4), the bending arm 14 is rotated by the motor, and the stirring roller 15 (and the rails 13 and the endless belt device 16) is moved downward to the second position. Be placed. In the second position, the lower end of the stirring roller 15 presses the culture vessel disposed on the loading table 11. Then, at the second position, by moving the culture vessel in the horizontal direction while pressing the culture vessel by the stirring roller 15, it is possible to appropriately stir the contents of the culture vessel.

  Next, as shown in FIG. 3 (5), the endless belt device 16 moves the stirring roller 15 to the stirring end position (moving end position) on the rail 13. The stirring end position is a position where the stirring roller 15 ends the movement on the front end of the culture vessel.

Next, as shown in FIG. 3 (6), the bending arm 14 is rotated by the motor, and the stirring roller 15 (and the rail 13 and the endless belt device 16) is moved upward to the first position. Be placed.
Next, the stirring roller 15 is moved by the endless belt device 16 again to the stirring start position shown in FIG. 3 (3).
And by repeating the process of FIG. 3 (3)-FIG. 3 (6), it is possible by the stirring roller 15 to stir a culture container continuously.

  The operation of the stirring device of the cell culture system of the present embodiment is not limited to this, and can be appropriately changed as long as the culture vessel can be continuously stirred. For example, after the state of FIG. 3 (6), the stirring roller 15 is moved to the position shown in FIG. 3 (2) (the first position and the rear end of the rail 13), and After confirmation, it is also possible to move to the stirring start position shown in FIG. 3 (3).

  As described above, according to the stirring device of the cell culture system of the present embodiment and the stirring method of the cell culture system, since the stirring roller 15 can be disposed on the rear outside of the loading table 11 in the initial state, It is possible to suppress the trouble of attaching and detaching the culture vessel to the cell culture system. In addition, since the rail 13 does not protrude from the surface of the loading platform 11, it is possible to reduce the risk of damaging the culture container when the culture container is attached to or removed from the cell culture system. .

  In addition, by making the height of the rail 13 changeable by the rotation of the bending arm 14 in this way, the stirring device can be realized with an inexpensive and simple structure without using an expensive slide rail or the like. It is possible.

Second Embodiment
Next, a second embodiment of the stirring device of the cell culture system of the present invention and the stirring method of the cell culture system will be described with reference to FIG. 4 and FIG.
As shown in FIG. 4, the stirring device 10 a of the cell culture system of the present embodiment is characterized in that clamps 17 are respectively provided at the rear end and the front end of the loading table 11. In the other points, the same configuration as the first embodiment can be provided to operate similarly.

The clamp 17 in the stirring device 10a of the cell culture system of the present embodiment is a fastener that clamps the culture container in the back and forth directions by sandwiching the rear end and the front end of the culture container placed on the loading table 11. It is.
Further, at this time, as shown in FIG. 5, it is preferable to set the clamping position of the clamp 17 (the height at which the culture container is clamped by the clamp 17) to a height approximately half the thickness of the culture container. By setting the clamping position of the clamp 17 in this manner, the thickness of the culture vessel can be kept more uniform.

Here, conventionally, when a large amount of culture medium is enclosed in the culture vessel, the thickness of the culture vessel is not kept constant, and the pressure in the culture vessel is the stirring roller when moving the stirring roller 15 on the upper surface of the culture vessel In some cases, the pressure exceeds the pressure of 15 and it becomes impossible to perform adequate stirring.
On the other hand, according to the stirring device 10a of the cell culture system of the present embodiment, the clamp 17 pulls the rear end portion and the front end portion of the culture container even when a large amount of culture medium is enclosed in the culture container. Thus, the thickness of the culture vessel can be kept substantially uniform, and the stirring roller 15 can be appropriately moved on the upper surface of the culture vessel.

In FIG. 4, the clamp 17 provided in front of the loading table 11 is composed of three parts. This is to arrange the port of the culture vessel and the tube connected to this between these three parts.
As described above, according to the stirring device 10a of the present embodiment, the operation of the stirring roller 15 can be stabilized.

[Cell culture system]
Next, the entire structure of the cell culture system provided with the stirring device of each of the above-described embodiments will be described in detail with reference to FIGS.
As shown in FIGS. 6 and 7, the cell culture system 1 is a system for storing a plurality of cell culture units U in multiple stages and performing cell culture individually for each of the cell culture units U. In FIG. 6, the cell culture system 1 has a two-tank structure in which the inside of the housing 2 is partitioned by the heat insulation wall 20 into an incubation tank 3 and a cold storage tank 4 that can be adjusted to a predetermined temperature. And the front doors 3a and 4a which can be opened and closed independently are provided in the incubation tank 3 and the cold storage tank 4, respectively.

  The cell culture unit U stored in the cell culture system 1 includes, for example, as shown in FIG. 7, a first culture container U20 and a second culture container U30 for use in cell culture, and these culture containers U20, A medium container U10 for supplying a medium to U30, a main tube U40 connecting the medium container U10 and the culture container U30, and a branch tube U42 connecting the culture container U30 and the main tube U40 it can.

The first culture vessel U20 is a culture vessel into which cells are first injected by injecting cells, and may be suitably used, for example, as an activated culture vessel used for cell culture for activating cells. it can. When activating suspended cells such as lymphocytes using the first culture vessel U20 as an activated culture vessel, a substance that activates cells such as an anti-CD3 antibody on the bottom of the first culture vessel U20 Is immobilized.
The second culture container U30 is a container for transferring the culture solution (cells and medium) in the first culture container after cell culture in the first culture container, and performing cell culture, for example, cells It can be suitably used as an amplification culture vessel or the like used for cell culture for proliferating.
Thus, by using the first culture container U20 as an activated culture container and using the second culture container U30 as an amplification culture container, suspension type cells such as lymphocytes can be suitably grown. is there.

  As shown in FIG. 7, these containers are connected to the main tube U40 in order of the culture medium container U10, the second culture container U30, and the first culture container U20. In addition, when it sees from the back side, it is connected to the main tube U40 in order of the 1st culture container U20, the 2nd culture container U30, and the culture container U10, and the order of these is the same.

In order to perform cell culture using the cell culture unit U configured in this way, first, cells are injected into the first culture container U20.
Next, the pump U41 is operated, and as shown in FIG. 7, the medium is transferred from the medium container U10 to the first culture container U20 by the pump U41 (the arrow (1) in the same figure). Then, cell culture is performed in the first culture container U20 for a predetermined period.
After the cell culture in the first culture vessel U20, the culture solution is transferred from the first culture vessel U20 to the second culture vessel U30 by the pump U41 (the arrow (2) in the same figure), and then from the medium vessel U10 The medium is transferred to the second culture vessel U30 (the arrow (3) in the same figure).

At this time, the transfer of the culture solution from the first culture container U20 to the second culture container U30 is performed via the branch tube U42 connected to the main tube U40 on the medium container U10 side of the pump U41.
In addition, the transfer of the culture medium from the culture medium container U10 to the second culture container U30 is performed via the branch tube U42 connected to the main tube U40 on the first culture container U20 side of the pump U41.

  According to such a cell culture unit U, transfer of the culture medium from the culture medium container U10 to the first culture container U20, transfer of the culture fluid from the first culture container U20 to the second culture container U30, and the culture medium The transfer of the three systems of transfer of the culture medium from the container U10 to the second culture container U30 can be appropriately performed by using only one pump U41.

  As the medium container U10, one having gas barrier properties against oxygen and carbon dioxide is generally used so that the pH of the stored medium does not change significantly during the culture period. In order to avoid that the high concentration of carbon dioxide gas contained in the culture medium escapes into the air and the concentration of carbon dioxide gas in the culture medium decreases and as a result, the pH rises, carbon dioxide is externally discharged from the inside of the culture medium container U10. It is desirable to minimize the leakage into the medium and to prevent the oxidation of the culture medium.

  The first culture container U20 and the second culture container U30 are formed in a bag shape (bag type) using a soft packaging material as a material, and some or all have transparency so that the contents can be confirmed. doing. In addition, these culture vessels are required to have gas permeability (permeability of oxygen and carbon dioxide) necessary for culturing cells, and are used in a culture environment of 37 ° C. and 5% carbon dioxide concentration. It is preferable to do. Furthermore, in order to realize high cell growth efficiency, it is preferable to have low cytotoxicity, low elution, and radiation sterilization suitability.

  As a material of the culture container which satisfy | fills such conditions, a polyethylene-type resin is preferable. Examples of the polyethylene-based resin include polyethylene, a copolymer of ethylene and α-olefin, a copolymer of ethylene and vinyl acetate, and an ionomer using ethylene and acrylic acid or a copolymer of methacrylic acid and metal ions. Moreover, polyolefin, a styrene-type elastomer, a polyester-type thermoplastic elastomer, a silicone-type thermoplastic elastomer, a silicone resin etc. can also be used.

The shapes of the culture medium container U10, the first culture container U20, and the second culture container U30 and the shapes of their containing portions are not particularly limited, and are rectangular in the example of FIG. These containers can be manufactured by sealing the four sides by heat sealing, and can also be manufactured as an integrally molded bag by blow molding.
Each port is preferably disposed on the main tube U40 side, and it is also preferable to make the accommodation portion gradually narrow toward the attachment portion of the tube.

  The materials of the main tube U40 and the branch tube U42 may be appropriately selected according to the use environment, but those having excellent gas permeability are desirable. For example, silicone rubber, soft vinyl chloride resin, polybutadiene resin, ethylene-vinyl acetate copolymer, chlorinated polyethylene resin, polyurethane thermoplastic elastomer, polyester thermoplastic elastomer, silicone thermoplastic elastomer, styrene elastomer, etc. are used. be able to. Further, as a styrene-based elastomer, for example, SBS (styrene-butadiene-styrene), SIS (styrene-isoprene-styrene), SEBS (styrene-ethylene-butylene-styrene), SEPS (styrene-ethylene-propylene-styrene), etc. Can be used.

In FIG. 7, for example, a silicone tube is used for the part to which the pump U41 is attached in the main tube U40, and a soft vinyl chloride resin tube can be suitably used for the other part.
Moreover, the connection between the tubes in the main tube U40 can be performed by a luer connector or other connection means. Furthermore, in the main tube U40, flow path opening / closing means such as a pinch valve, a two-way stopcock, a three-way stopcock, etc. may be provided.

  The type of pump U41 is not particularly limited, but a tube roller pump such as Perister pump (R) can be suitably used because a closed system in the cell culture unit U can be easily realized.

  The cell culture system 1 stores such a cell culture unit U, but the first culture container U20 and the second culture container U30 are maintained at a temperature (eg, 37 ° C.) suitable for cell culture. The medium container U10 is stored in the temperature-controlled cold storage tank 4 so as to be maintained at a temperature (for example, 4 ° C.) suitable for storing the culture medium.

  As shown in FIG. 8, in the incubation chamber 3 in which the first culture container U20 and the second culture container U30 are stored, a plurality of slide rails 31 are arrayed in multiple stages along the height direction. A side support frame 30 is installed. A culture container housing shelf 32 for housing the first culture container U20 and the second culture container U30 is slidably attached to each of the slide rails 31 arranged in a row on the incubation tank side support frame 30. It can be pulled out to the front. By doing this, when attaching or detaching the first culture container U20 and the second culture container U30 to the culture container housing shelf 32, the culture container housing shelf 32 is pulled out to the front, It is designed to make it easy to work.

The stirring device of the cell culture system of the embodiment described above can be disposed on each culture container housing shelf 32. Also, it may be provided for each one of the first culture container U20 and the second culture container U30, or for both of them. When the first culture vessel U20 is used as an activated culture vessel and the second culture vessel U30 is used as an amplification culture vessel in order to suitably proliferate floating cells such as lymphocytes, the stirring device is It may be provided only in the second culture vessel U30.
In the example shown in FIG. 8, the pump U41 and the closing member constituting the cell culture unit U are attached to the front side of the culture vessel housing shelf 32, but the illustration of the main tube U40 including these is omitted. Yes.

  Similarly, in the cold storage tank 4 in which the culture medium container U10 is stored, a cold storage tank side support frame 40 in which a plurality of slide rails 41 are arranged in multiple stages along the height direction is installed. A culture medium container storage rack 42 for storing the culture medium container U10 is slidably attached to each of the slide rails 41 arranged in a row on the cold storage tank side support frame 40 and can be pulled out to the front. Thus, even when the culture medium container U10 is attached to or detached from the culture medium container accommodation rack 42, the culture medium container accommodation rack 42 is pulled out to the front so that these operations can be easily performed.

  As described above, in the cell culture system 1, the culture vessel housing shelf 32 and the culture medium container housing shelf 42 are separately provided in pairs in the incubation tank 3 and the cold storage tank 4 adjusted to a predetermined temperature, The first culture container U20 and the second culture container U30 are housed in the culture container housing shelf 32, and the culture container U40 is housed in the culture container housing shelf 42, so that a plurality of cell culture units U can be multistaged. It can be stored in By doing this, according to the cell culture system 1, the plurality of cell culture units U can be independently operated under the temperature environment suitable for each of the first culture container U20 and the second culture container U30 and the medium container U10. In this state, the cells can be stored in multiple stages so that cell culture can be performed individually for each cell culture unit U.

A schematic cross-sectional view of the culture vessel housing shelf 32 as viewed from the front side is shown in FIG. In FIG. 9, the first culture vessel U20 is omitted.
As shown in the figure, the content of the second culture container U30 can be stirred by reciprocating the powder while pressing the stirring roller 15 against the upper surface of the second culture container U30.

Further, as described above, in the initial state, the stirring roller 15 is disposed outside the loading stand 11, and the rails 13 for guiding the stirring roller 15 are also disposed outside the loading stand 11, and the upper surface of the rail 13 is , And the upper surface of the loading base 11 are disposed substantially on the same plane. For this reason, when the culture vessel is attached to or removed from the loading stand 11, the stirring roller 15 and the rails 13 do not disturb the culture vessel, and the risk of damaging the culture vessel can be reduced. ing.
In particular, in a cell culture system in which a plurality of cell culture units are thus stored in a multistage manner in a housing and cell culture is performed individually for each cell culture unit, it is extremely easy to safely and easily attach and remove the culture container. Important to you.

Although the present invention has been described with reference to the preferred embodiments, it is needless to say that the present invention is not limited to the above-described embodiments, and various modifications can be made within the scope of the present invention.
For example, in each embodiment, the stirring roller 15 is moved while being pressed and pushed from the rear to the front, but on the contrary, it is moved while being pressed and moved from the front to the rear and stirred, or while being pushed in both forward and backward directions. It is possible to change suitably, such as moving and stirring.

  The present invention can be suitably used in regenerative medicine, immunotherapy, antibody drug production and the like in which cells are cultured in large amounts in a closed system.

DESCRIPTION OF SYMBOLS 1 cell culture system 2 housing | casing 10, 10a stirring apparatus 11 loading stand 12 base 13 rails 14 bending | flexion arm 15 stirring roller 16 endless belt apparatus 17 clamp 20 heat insulation wall 3 incubation tank 30 incubation tank side support frame 31 slide rail 32 culture Container storage rack 4 Cold storage tank 40 Cold storage tank side support frame 41 Slide rail 42 Medium container storage rack U Cell culture unit U10 Medium container U20 First culture container (culture container)
U30 Second culture vessel (culture vessel)

Claims (7)

A stirring apparatus for a cell culture system, which performs cell culture using a culture vessel containing a medium with cells.
A loading platform on which the culture vessel is placed;
A base supporting the loading platform,
An elongated stirring member movable horizontally while pressing the upper surface of the culture vessel;
Two movably supported both ends of the stirring member, and provided horizontally on both opposite left and right sides of the loading base, the length being longer than the width in the front-rear direction of the loading base and vertically movable. With the rails,
Two pairs of arms rotatably coupled to the respective rails and the base for moving the rails vertically while maintaining the horizontal state;
First driving means for pivoting the two sets of arms around a joint between the two sets of arms and the base;
And d) a second drive means for moving the stirring member in the longitudinal direction on the two rails;
One end of the rail extends to the outside of the width in the front-rear direction of the loading table in a state where the upper surface of the rail is disposed substantially in the same plane as the upper surface of the loading table , and the stirring member A stirrer for a cell culture system, which is placed outside the pedestal.   The stirring device of the cell culture system according to claim 1, wherein the two sets of arms comprise two arms connected to the front and the rear of the rail, respectively.   The stirring device of the cell culture system according to claim 1 or 2, wherein the stirring member is a roller.   The stirring device of the cell culture system according to any one of claims 1 to 3, further comprising: a fastener for holding the end of the culture vessel at the front end and the rear end of the loading table.   The cell culture system according to any one of claims 1 to 4, characterized in that the height at which the end of the culture vessel is sandwiched by the fastener is approximately half the thickness of the culture vessel. Stirring device. The cell culture system is
A cell culture system that stores a plurality of cell culture units including at least the culture container and a culture medium container that supplies a culture medium to the culture container in multiple stages in a housing, and performs cell culture individually for each of the cell culture units. A stirring device is provided for each of the culture vessels,
An incubation tank in which the culture container is stored, and a cold storage tank in which the culture medium container is stored,
An incubation tank side support frame and a cold storage tank side support frame in which a plurality of slide rails are arranged in multiple stages along the height direction are installed in each of the incubation tank and the cold storage tank,
A culture container housing shelf for housing the culture containers is slidably attached to each of the slide rails arranged in multiple stages in the incubation tank side support frame, and is arranged in multiple stages in the cold storage tank side support frame. A culture medium container storage shelf for housing the culture medium container is slidably attached to each of the slide rails.
The cell according to any one of claims 1 to 5, wherein the culture vessel storage rack and the culture medium storage rack are separately provided in pairs in the incubation tank and the cold storage tank. Stirring device for culture system. A method of stirring a cell culture system by the stirrer of the cell culture system according to any one of claims 1 to 6,
(A) The first driving means can horizontally move the upper part of the culture vessel without pressing the culture vessel from the state where the stirring member is disposed outside the loading table Turn the arm to move the rail upwards until it is placed in the first position,
(B) the second driving means moves the stirring member to a stirring start position on the rail;
(C) the first driving means is at the second position where the stirring member can move the upper surface of the culture vessel horizontally while the stirring member presses the culture vessel from the first position Rotate the arm to move the rail downward until it is deployed,
(D) the second driving means moves the stirring member to the stirring end position on the rail;
(E) the first driving means rotates the arm to move the rail upward such that the stirring member is disposed at the first position from the second position;
Subsequently, the operations of (b) to (e) are repeated, and the method of stirring the cell culture system.

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