JP5254319B2 - Multi-axis rotary agitator and method for producing cultured cells using the same - Google Patents

Description

  The present invention relates to a multi-axis rotary stirring device and a method for producing cultured cells using the same.

  As a multi-axis rotating device, a rotating device that rotates a container by n-axis (n is an integer of 2 or more), and a fluid supply that supplies fluid to the container from a stationary system while the container is rotated by n-axis A three-dimensional clinostat provided with a device is known (Patent Document 1).

JP 2003-70458 A

In the conventional multi-axis rotating device, the inside of the container can be set in a pseudo-gravity state, but there is a problem that the stirring efficiency inside the container is remarkably lowered.
An object of the present invention is to provide a stirring device that can change the inside of a container from a pseudo microgravity state to a hypergravity state and that is excellent in stirring efficiency.

  In the present invention, “stirring” means including shaking. In addition, the “pseudo microgravity state” means a pseudo-weightless state (a state close to a state where there is no influence of gravity), and a state close to a state where the influence of gravity is slight although it does not go to zero gravity. means. Further, the “overgravity state” means a state in which there is a stronger influence than gravity due to centrifugal force or the like.

A feature of the multi-axis rotary stirring device of the present invention is that a rotating means (50) for rotating the container (40) about the first axis (10) as a central axis,
Rotating means (60) for rotating the container (40) about a second axis (20) intersecting the first axis (10);
The first axis (10) and the second axis (20) are rotated by rotating the first axis (10) with the third axis (30) orthogonal to the first axis (10) and the second axis (20) as a rotation axis. Crossing angle changing means (70) for changing the crossing angle of
The point is that the container (40) is held at a position offset from the third axis (30).

  The rotating means (50) for rotating the container (40) about the first axis (10) is not limited as long as the container (40) can be rotated, and known rotating means can be applied. For example, the rotation of a motor (a device that converts energy such as electricity, steam, gasoline, or rocket fuel into a mechanical rotary motion and transmits it. An electric motor is preferable from the viewpoint of simplicity) directly or indirectly ( Means for rotating the container (via gears, belts, chains, etc.) are included. That is, the rotating means (50) includes a motor and a transmission tool (shaft, gear, belt, etc.).

  If the rotation means (60) for rotating the container (40) around the second axis (20) intersecting the first axis (10) can rotate the container (40) together with the first axis, there is no limitation. In addition, a known rotating means can be applied. For example, a rotating means similar to the rotating means (50) is included.

  The first axis (10) and the second axis (20) are mechanically coupled, and even when the second axis (20) is rotated at a high speed, the first axis (10) and the second axis ( 20). If the first axis (10) and the second axis (20) are not separated from each other, it is possible to more easily give the container (40) a complicated operation and a variety of operations.

  As an example in which the first shaft (10) and the second shaft (20) are separated from each other, for example, two rotating rolls that rotate in the same direction are used as the rotating means (50). The case where the first shaft (10) is provided so as to coincide with the rotation axis of the container (40) is included. That is, the present invention includes such a case. In such a case, the container (40) is separated from the rotating roll as the rotation of the second shaft (20) is increased. Because one axis (10) will be away from the second axis (20)}, there is some restriction on the motion imparted to the container (40).

  The rotating means (50) and the rotating means (60) may be rotating means caused by the same motor (common motor), or may be rotating means caused by separate motors.

  The rotation means (50) and / or the rotation means (60) may be continuous rotation, intermittent rotation, or a combination of forward rotation and reverse rotation.

  The crossing angle changing means (70) for changing the crossing angle between the first axis (10) and the second axis (20) is a third that is orthogonal to the first axis (10) and the second axis (20). There is no limitation as long as the crossing angle between the first axis (10) and the second axis (20) can be changed by rotating the first axis (10) about the axis (30) as a rotation axis, and a known crossing angle. Change means can be applied. For example, in addition to the rotating means similar to the rotating means (50), there is a means for extending / stretching or bending the third shaft (30) so as to become a rotating shaft by an actuator (solenoid, feed screw, hydraulic cylinder, pneumatic cylinder, etc.) or the like. included.

  The intersecting angle changing means (70) may be a continuous change, an intermittent change, or a combination of normal rotation and reverse rotation.

  The crossing angle (degree) between the first axis (10) and the second axis (20) is preferably 0 to 90, more preferably 10 to 90, next preferably 20 to 90, and next preferably 30 to 30. 90, particularly preferably 40-90. The intersection angle means a small angle among the angles formed by the intersection of the first axis and the second axis (for example, 90 degrees when the formed angle is 90 degrees, and similarly 80 80 degrees for 100 degrees and 100 degrees, and similarly 10 degrees for 10 degrees and 170 degrees).

  The container (40) is held at a position offset from the third axis (30). The container (40) may be held at a position offset from both the second axis (20) and the third axis (30), or the first axis (10) and the third axis (30). It may be held at a position offset from any of them, and may be held at a position offset from any of the first axis (10), the second axis (20), and the third axis (30). . The container (40) is preferably held at a position offset from both the second axis (20) and the third axis (30).

  The offset means that the reference line (the first axis (10), the second axis (20) and / or the third axis (30)) is apart from the reference line by a certain distance. May be held on the first axis (10) or on the second axis (20), but held on the intersection of the first axis (10), the second axis (20) and the third axis (30). Is not.

  The second axis (20) is fixed horizontally to the ground surface, the third axis (30) is orthogonal to the second axis (20), and the first axis (10) is relative to the second axis (20). It is preferable that they intersect.

  Although there is no restriction | limiting in the number of containers (40), 1-20 pieces are preferable, More preferably, it is 1-12 pieces. When there is one container (40), it is preferable to hold the container (40) so that the center of gravity of the container is on the first axis (10). On the other hand, when there are a plurality of containers (40), the containers are arranged around the first axis (10) (see FIGS. 7 to 9), and the center of gravity of the plurality of container groups is the first axis (10). It is preferable to hold the container group so as to be on top.

  There is no restriction | limiting in the shape of a container (40), a magnitude | size, and a material, Each can be suitably determined according to a use etc. When applied to cell culture or the like to be described later, it is a transparent container or translucent container (material: polyethylene, polypropylene, polymethyl methacrylate, polystyrene, polycarbonate, cycloolefin polymer, polyethersulfone or glass) for observing the inside. It is preferable.

  It is preferable that the position of the first shaft (10) does not vary depending on the number, shape, size, and material of the container (40). If the position of the first axis (10) does not fluctuate in this way, it is possible to more easily give the container (40) a complicated operation and a variety of operations.

  As an example in which the position of the first shaft (10) varies depending on the number, shape, size, and material of the container (40), for example, two rotating rolls that rotate in the same direction are used as the rotating means (50). The case where the container (40) is placed on the two rotating rolls and the first shaft (10) is provided so as to coincide with the rotation axis of the container (40) is included. The present invention includes such a case. In such a case, when the size of the container (40) is changed so that the turning radius changes, the position of the first shaft (10) is changed. Therefore, there is a certain limit to the operation given to the container (40).

  It is preferable that the multiaxial rotary stirring device of the present invention further includes a fluid supply / discharge tool (80) for taking fluid into and out of the container (40). When the fluid supply / discharge tool (80) is provided, when the multi-axis rotating stirrer of the present invention is applied to cell culture or the like to be described later, the container (40) is kept in a rotating and stirring state while fluid is contained in the container (40). (Culture liquid, carbon dioxide gas, cell dispersion, and a mixture thereof) can be supplied, and the fluid can be discharged from the container (40) while the container (40) is kept in a rotationally stirred state. (This supply and discharge can be done in parallel or independently.)

  The fluid means a fluid body, and includes a liquid, a gas, a supercritical fluid, a mixture (homogeneous or heterogeneous) thereof, and a dispersion in which a solid is dispersed.

  The fluid supply / discharge tool (80) can be used without limitation as long as it is a device that can draw fluid into and out of the container (40) while keeping the container (40) in a rotationally stirred state. For example, <1> rotor Combination of Lee joint, piping and pump, etc. <2> Fluid storage container rotated with container (40) (carbon dioxide cylinder etc.), Combination of piping and valve, etc. <3> Fluid storage container rotated with container (40) , Combinations of empty containers, piping and pumps that rotate with the container (40), and combinations thereof.

  The multiaxial rotary stirring device of the present invention can be applied to mixing various materials and the like, and is suitable for cell culture. Furthermore, when applied as a multi-axis rotary agitator for cell culture, as described above, the multi-axis rotary agitator further includes a fluid supply / discharge tool (80) for taking fluid into and out of the container (40). It is preferable to provide. When the fluid supply / discharge tool (80) is provided, the fluid (culture solution, carbon dioxide gas, cell dispersion, mixture thereof, etc.) is supplied into the container (40) while keeping the container (40) in a rotationally stirred state. In addition, the fluid can be discharged from the container (40) while keeping the container (40) in a rotating and stirring state (this supply and discharge can be performed in parallel or independently). .

  The multi-axis rotation stirring device of the present invention may be connected to a control device (multi-axis controller, PLC (programmable logic controller), arithmetic device, etc.) for automatic control. When connected to a control device, more complex operations can be easily performed, and the same conditions can be easily reproduced. Therefore, when connected to the control device, the quality of the results (product or test data, etc.) obtained by production or testing using the multi-axis rotary stirring device of the present invention is further improved. In addition, production or testing can be performed more efficiently.

  The multi-axis rotating stirrer of the present invention includes a temperature sensor, a pressure sensor, a temperature controller (cooling, heating), a CCD camera, a lighting device (visible light, ultraviolet light, infrared light, etc.), a voltage / current supply device, An electromagnetic wave supply device or the like can be provided.

  When a temperature sensor and / or a pressure sensor are provided, it is preferable to provide a temperature and pressure in the container (40).

  When providing a temperature regulator (cooling, heating), it is preferable to provide so that the temperature in a container (40) can be adjusted. The temperature controller may directly heat / cool the container (40), or may heat / cool the heating medium and indirectly heat / cool the container (40). The temperature regulator is used with a temperature sensor. When a temperature controller and a temperature sensor are provided, cell culture can be performed without using a cell culture incubator.

  When providing a CCD camera, it is preferable to provide it so that the state in the container (40) can be observed. The CCD camera may be either a high speed CCD or an infrared CCD.

  When providing an illuminating device (visible light, ultraviolet light, infrared light, etc.), it is preferable to provide it so that the inside of a container (40) can be illuminated. The illuminator can be used as an illumination of a CCD camera, a light source for cell culture of plant cells, or a light source of a photochemical reaction (including the case of using a photoinitiator).

  When the voltage / current supply device and / or the electromagnetic wave supply device are provided, it is preferable to provide the container (40) so that the voltage / current and / or electromagnetic wave can be applied.

The feature of the method for producing cultured cells of the present invention is summarized in that it includes a step of culturing cells using the above-described multiaxial rotary stirring device.
As the culture solution, scaffold, cells and the like, those used in normal cell culture can be used (hereinafter the same).

  The features of the method for producing cultured cells of the present invention include the step of culturing cells using the above-described multi-axis rotating stirrer provided with a fluid supply / discharge tool (80) for taking fluid into and out of the container (40), In this cell culturing step, the gist is that the cells are collected in the vicinity of the vessel wall of the container (40) by centrifugal force while rotating the container (40) and the culture solution is exchanged.

  When exchanging the culture solution in this way, the cells are collected near the vessel wall of the container (40) by the centrifugal force generated by increasing the rotation speed of the first shaft (10) or the second shaft (20), It is preferable to discharge the culture solution near the rotating shaft and supply a new culture solution (see FIG. 10). This supply and discharge can be done in parallel or independently.

The culture medium may be exchanged continuously or intermittently at regular intervals. Moreover, the kind of culture solution can also be changed in the middle of cell culture.
Note that it is not necessary to replace all of the culture broth to be replaced by the above method, and the culture broth may be replaced by removing the container (40) from the multi-shaft rotary stirring device.

  The feature of the method for producing cultured cells of the present invention includes the step of culturing cells using the above-described multi-axis rotating stirrer. In this step of culturing cells, the gas in the vessel is exchanged while rotating the vessel (40). The point to do is a summary.

  When gas is exchanged in this way, the space filled with gas is always maintained while the container (40) is rotating by adjusting the inclination of the first axis (10) and / or the third axis (30). It is preferable to create and discharge gas from this space and supply new gas (see FIG. 11). This supply and discharge can be done in parallel or independently.

The exchange of gas may be performed continuously or intermittently at regular intervals. In addition, the type of gas can be changed during cell culture.
In addition, it is not necessary to replace | exchange all the gas to replace | exchange by said method, A container (40) may be removed from a multiaxial rotation stirring apparatus, and gas may be replaced | exchanged.

  The feature of the method for producing cultured cells of the present invention includes the step of culturing cells using the multi-axis rotating stirrer. In this step of culturing cells, the cells in the vessel are discharged while rotating the vessel (40). The point to do is a summary.

  When the cells are discharged in this way, the tilt of the first axis (10) and / or the third axis (30) is adjusted, and the container (40) is always filled with the culture medium containing the cells while the container (40) is rotating. It is preferable to create a space and discharge a culture solution containing cells from the space and supply a gas (see FIG. 12). This supply and discharge can be done in parallel or independently.

The cells may be discharged continuously or intermittently at regular intervals.
In addition, it is not necessary to discharge | emit all the cells by said method, A container (40) may be removed from a multi-axial rotation stirring apparatus, and cells may be discharged | emitted.

  The multiaxial rotary stirring device of the present invention can change the inside of a container from a pseudo microgravity state to an overgravity state, and exhibits excellent stirring efficiency.

  The multi-axis rotary stirring device of the present invention can exhibit various stirring operations. For example, by reciprocally rotating the container (40) held at a position offset from both the second axis (20) and the third axis (30) using the second axis (20) as a rotation axis, the container (40) can be reciprocated (reciprocating) (FIG. 3). Further, the container (40) held at a position offset from both the second axis (20) and the third axis (30) is reciprocally rotated about the second axis (20) as a rotation axis, thereby (40) can be rocked (seesaw) (FIG. 4). Further, the container (40) is rotated by rotating the container (40) held on the first shaft (10) (or the container (40) held at a position offset from the first shaft (10)). It can be moved (rotated) (FIG. 5). Moreover, the exercise | movement which combined these can be made. Furthermore, each of these movements can be repeated or randomly combined, and the movements can be programmed by the control device.

  When a fluid supply / discharge tool (80) for taking fluid into and out of the container (40) is provided, fluid {liquid (culture solution etc.), gas (carbon dioxide gas etc.) and slurry (cell dispersion etc.)} It can supply in a container (40), maintaining a rotation stirring state. Further, the fluid {liquid (culture solution, etc.), gas (carbon dioxide gas, etc.) and slurry (cell dispersion, etc.)} can be discharged from the container (40) while maintaining a rotating stirring state (this supply). And discharging can be done in parallel or independently.)

  When the number of containers (40) is plural, a plurality of kinds of stirring operations can be performed at once, the center of gravity of the container (40) is on the first axis (10), and the first The container (40) can be held at a position offset from any of the axis (10), the second axis (20), and the third axis (30).

  The multi-axis rotating stirrer of the present invention is suitable for cell culture because it can change the inside of a container from a pseudo microgravity state to an overgravity state and exhibits excellent stirring efficiency. In the field of regenerative medicine, even if the shape of the scaffold (such as a blood vessel tubular scaffold or an organ scaffold) is complex, the use of the multi-axis rotating stirrer of the present invention significantly improves the stirring efficiency. By following complicated shapes, excellent cell culture can be realized.

  In the method for producing cultured cells of the present invention, since cell culture is performed using the above-described multiaxial rotary stirring device, cell culture can be performed from a pseudo microgravity state to an overgravity state. Moreover, about the cell (plant cell etc.) which feels gravity, it can culture by changing the influence of gravity.

  The method for producing cultured cells according to the present invention uses the multi-axis rotating stirrer provided with the fluid supply / discharge tool (80) for taking fluid into and out of the container (40), so that the container (40) is rotated. However, it is easy to collect the cells near the vessel wall of the container (40) by centrifugal force and exchange the culture solution.

  The method for producing cultured cells according to the present invention uses the multi-axis rotating stirrer provided with the fluid supply / discharge tool (80) for taking fluid into and out of the container (40), so that the container (40) is rotated. However, the gas in the container can be easily exchanged.

  The method for producing cultured cells according to the present invention uses the multi-axis rotating stirrer provided with the fluid supply / discharge tool (80) for taking fluid into and out of the container (40), so that the container (40) is rotated. However, the cells in the container can be easily discharged.

  Therefore, when the method for producing cultured cells of the present invention is applied, cells can be continuously cultured, and a large number of cells can be produced from a small amount. In the field of regenerative medicine, even if the shape of the scaffold (such as a tubular scaffold for blood vessels, an scaffold for organs, etc.) is complex, if the method for producing cultured cells of the present invention is used, the stirring efficiency is remarkably excellent. By following complicated shapes, excellent cell culture can be realized.

  Hereinafter, the multiaxial rotary stirring device of the present invention will be described in more detail with reference to the drawings. Unless otherwise specified, the items described first can be applied in common to the description of subsequent drawings.

<Multi-axis rotation stirring apparatus of FIGS. 1 and 2>
FIG. 1 is a side view schematically showing one embodiment of the multi-axis rotary stirring device of the present invention.
FIG. 2 is a top view schematically showing one embodiment of the multi-axis rotation stirring device of the present invention (a top view corresponding to the side view of FIG. 1).

  An electric motor (51) and a bevel gear (52) are arranged as rotating means (50) for rotating the container (40) about the first axis (10) as a central axis. The rotation of the electric motor (51) is transmitted to the container (40) via the bevel gear (52), and the container (40) rotates about the first axis (10).

  An electric motor (61) and a gear (62) are arranged as rotating means (60) for rotating the container (40) around a second axis (20) intersecting the first axis (10). The rotation of the electric motor (61) is transmitted to the container (40) via the gear (62), and the container (40) rotates about the second axis (20). Along with this rotation, the first shaft (10) and the third shaft (30) also rotate.

  The first axis (10) and the second axis (20) are rotated by rotating the first axis (10) with the third axis (30) orthogonal to the first axis (10) and the second axis (20) as a rotation axis. An electric motor (71) is arranged as a crossing angle changing means (70) for changing the crossing angle. The rotation of the electric motor (71) is directly transmitted to the container (40), the electric motor (51) and the bevel gear (52), and the container (40) and the electric motor (51) are centered on the third axis (30). And the bevel gear (52) rotates, and the crossing angle between the first axis (10) and the second axis (20) changes.

  The container (40) is held on the first axis (10) and at a position offset from both the second axis (20) and the third axis (30).

<Multi-axial rotary stirring device in FIG. 6>
FIG. 6 is a perspective view schematically showing one aspect of the multi-axis rotary stirring device of the present invention.

  An electric motor (51) is arranged as a rotating means (50) for rotating the container (40) about the first axis (10). The rotation of the electric motor (51) is directly transmitted to the container (40), and the container (40) rotates around the first axis (10).

  An electric motor (61) is arranged as a rotating means (60) for rotating the container (40) around the second axis (20) intersecting the first axis (10). The rotation of the electric motor (61) is transmitted to the container (40) via the frame (90), and the container (40) rotates about the second axis (20). Along with this rotation, the first shaft (10) and the third shaft (30) also rotate.

  The first axis (10) and the second axis (20) are rotated by rotating the first axis (10) with the third axis (30) orthogonal to the first axis (10) and the second axis (20) as a rotation axis. As a cross angle changing means (70) for changing the cross angle, an actuator (feed screw or the like) is disposed in the frame (90). The force of the actuator is transmitted to the container (40) and the electric motor (51), and the container (40) and the electric motor (51) are rotated about the third axis (30) as the central axis, thereby the first axis (10). And the intersection angle of the second axis (20) changes.

  The container (40) is held on the first axis (10) and at a position offset from both the second axis (20) and the third axis (30).

<Multi-axial rotary stirring device in FIG. 13>
FIG. 13 is a partial front view schematically showing one embodiment of the multi-axis rotary stirring device of the present invention.

  An electric motor (51) is arranged as a rotating means (50) for rotating the container (40) about the first axis (10). The rotation of the electric motor (51) is directly transmitted to the container (40), and the container (40) rotates around the first axis (10).

  An electric motor (61) is arranged as a rotating means (60) for rotating the container (40) around the second axis (20) intersecting the first axis (10). The rotation of the electric motor (61) is transmitted to the container (40) via the feed screw (72) and the like, and the container (40) rotates about the second axis (20). Along with this rotation, the first shaft (10) and the third shaft (30) also rotate.

  The first axis (10) and the second axis (20) are rotated by rotating the first axis (10) with the third axis (30) orthogonal to the first axis (10) and the second axis (20) as a rotation axis. An electric motor (71), a feed screw (72), a moving nut (73), and a slider (74) are arranged as crossing angle changing means (70) for changing the crossing angle. The rotation of the electric motor (71) is converted into a linear movement of the moving nut (73) through the rotation of the feed screw (72), and the slider (74) moves linearly on the second axis (20) along with this movement. As a result, the container (40) and the electric motor (51) rotate about the third axis (30) as the central axis, and the crossing angle between the first axis (10) and the second axis (20) changes.

  In the multi-axis rotary stirring device of FIG. 13, the feed screw (72) and the moving nut (73) may be other actuators. Alternatively, the slider (74) may be changed to an actuator, or the slider (74) may be changed to an actuator, and the electric motor (71), the feed screw (72) and the moving nut (73) may be changed to the slider (74). Also good.

The third axis (30) exists as a line perpendicular to the drawing at the point where the first axis (10) and the second axis (20) intersect in FIG.
The moving nut (73) is connected to the first shaft (10) and the link mechanism (75) so that the crossing angle between the first shaft (10) and the second shaft (20) can be freely changed, and a feed screw ( 72), it can be moved linearly on the feed screw (72).
The slider (74) is connected to the first shaft (10) and the link mechanism (75) so that the intersection angle between the first shaft (10) and the second shaft (20) can be freely changed, and the second shaft ( 20) It is configured so that it can freely move in a straight line.

  The container (40) is held on the first axis (10) and at a position offset from both the second axis (20) and the third axis (30).

It is the side view which showed typically the one aspect | mode of the multi-axial rotation stirring apparatus of this invention. It is the top view which showed typically the one aspect | mode of the multi-axial rotation stirring apparatus of this invention (it is a top view corresponding to the side view of FIG. 1). It is the conceptual diagram which showed typically the reciprocating (reciprocating) operation | movement among the stirring operation | movement of the multi-axial rotation stirring apparatus of this invention (the lower side of drawing is a gravity direction). It is the conceptual diagram which showed typically rocking (seesaw) operation | movement among the stirring operation | movement of the multi-axial rotation stirring apparatus of this invention (the lower side of drawing is a gravity direction). It is the conceptual diagram which showed typically rotary (rotation) operation | movement among the stirring operation | movement of the multi-axial rotation stirring apparatus of this invention (the lower side of drawing is a gravity direction). It is the perspective view which showed typically the one aspect | mode of the multi-axial rotation stirring apparatus of this invention. It is the perspective view which showed typically the one aspect | mode of the arrangement | positioning of the container in the case of using a several container in the multi-axial rotation stirring apparatus of this invention. It is the perspective view which showed typically the one aspect | mode of the arrangement | positioning of the container in the case of using a several container in the multi-axial rotation stirring apparatus of this invention. It is the perspective view which showed typically the one aspect | mode of the arrangement | positioning of the container in the case of using a several container in the multi-axial rotation stirring apparatus of this invention. In the step of culturing cells using the multi-axis rotating stirrer of the present invention provided with a fluid supply / discharge tool (80) for taking fluid into and out of the vessel (40), the cells are put into the vessel by centrifugal force while rotating the vessel. It is the partial front view which showed typically a mode that it collects in the vicinity of the container wall of this, and replace | exchanges a culture solution. In the step of culturing cells using the multi-axis rotary stirring device of the present invention provided with a fluid supply / discharge tool (80) for taking fluid in and out of the vessel (40), the gas in the vessel is exchanged while rotating the vessel. It is the partial front view which showed the mode to do typically. In the step of culturing cells using the multi-axis rotating stirrer of the present invention provided with a fluid supply / discharge tool (80) for taking fluid in and out of the container (40), the cells in the container are discharged while rotating the container. It is the partial front view which showed the mode to do typically. It is the partial front view which showed typically the one aspect | mode of the multi-axial rotation stirring apparatus of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 1st axis | shaft 20 2nd axis | shaft 30 3rd axis | shaft 40 Container 41 Container 42 Container 43 Container 44 Container 45 Container 46 Container 47 Container 48 Container 49 Container 51 Electric motor 52 Bevel gear 61 Electric motor 62 Gear 71 Electric motor 72 Feed screw 73 Moving nut 74 Slider 75 Link mechanism 80 Fluid supply / discharge tool 90 Frame 100 Holder 110 Cell 120 Gas

Claims (8)

A rotating means (50) for rotating the container (40) about the first axis (10) as a central axis;
Rotating means (60) for rotating the container (40) about a second axis (20) intersecting the first axis (10);
The first axis (10) and the second axis (20) are rotated by rotating the first axis (10) with the third axis (30) orthogonal to the first axis (10) and the second axis (20) as a rotation axis. Crossing angle changing means (70) for changing the crossing angle of
And a container (40) held at a position offset from the third shaft (30). The multi-shaft rotary stirring device according to claim 1, further comprising a fluid supply / discharge tool (80) for taking fluid into and out of the container (40). The multi-axis rotary stirring device according to claim 1 or 2, wherein the number of containers (40) is 1 to 20. The multiaxial rotary stirring device according to any one of claims 1 to 3, which is used for cell culture. A method for producing cultured cells, comprising a step of culturing cells using the multiaxial rotary stirring device according to claim 1. A step of culturing cells using the multi-axis rotating stirrer according to claim 2, wherein in the step of culturing cells, the cells are moved near the wall of the vessel (40) by centrifugal force while rotating the vessel (40). A method for producing cultured cells, characterized in that the culture solution is exchanged after collecting them. A cell culture step comprising the step of culturing cells using the multi-axis rotating stirrer according to claim 2, wherein the gas in the vessel is exchanged while rotating the vessel (40) in the cell culturing step. Cell production method. A cell culture step comprising the step of culturing cells using the multi-axis rotating stirrer according to claim 2, wherein the cells in the vessel are discharged while rotating the vessel (40) in the cell culturing step. Cell production method.

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