Bioreactor, bioreactor system, and method for operating bioreactor

Abstract

A bioreactor according to the present disclosure comprises: an outer cylinder which is centered on an axis line extending in the vertical direction; an inner cylinder which is provided inside the outer cylinder and centered on the axis line; a rotary cylinder which is disposed between the outer cylinder and the inner cylinder, defines a first pool between the inner cylinder and the rotary cylinder, defines a second pool between the outer cylinder and the rotary cylinder, and is rotatable about the axis line; a drive unit for rotationally driving the rotary cylinder around the axis line; a first supply port through which a liquid is supplied to the first pool; and a second supply port through which a liquid is supplied to the second pool.

Description

Bioreactor, bioreactor system, and method of operating a bioreactor

[0001] This application claims priority to Japanese Patent Application No. 2023-108838, filed on June 30, 2023, the contents of which are incorporated herein by reference.

[0002] For example, Patent Document 1 discloses a bioreactor for culturing cells. This bioreactor has a double-cylinder structure comprising a rotating cylinder and an outer cylinder. A pool for containing a culture solution is partitioned and formed between the rotating cylinder and the outer cylinder. When the rotating cylinder rotates, Taylor vortices are formed in the culture solution contained in the pool. The Taylor vortices agitate the culture solution, promoting cell culture.

[0003] Japanese Patent Application Publication No. 8-308560

[0004] The environmental conditions suitable for culturing cells change depending on the growth phase of the cells. In particular, it is preferable to change the components of the culture medium that serves as the cell growth environment depending on the growth phase. However, when using the bioreactor described in Patent Document 1, it is necessary to install multiple different bioreactors according to the cell growth stage, which increases the installation space.

[0005] The present disclosure provides a bioreactor, a bioreactor system, and a method for operating a bioreactor that are capable of culturing cells according to their growth phase in a compact configuration.

[0006] The bioreactor according to the present disclosure comprises an outer cylinder centered on an axis extending in the vertical direction, an inner cylinder centered on the axis inside the outer cylinder, a rotating cylinder disposed between the outer cylinder and the inner cylinder, defining a first pool between the outer cylinder and the inner cylinder and defining a second pool between the outer cylinder and the inner cylinder, and rotatable about the axis, a drive unit that drives the rotating cylinder to rotate about the axis, a first supply port capable of supplying liquid to the first pool, and a second supply port capable of supplying liquid to the second pool.

[0007] The bioreactor system according to the present disclosure comprises the above-described bioreactor and a control device that controls the bioreactor, wherein the bioreactor comprises a first culture medium supply unit capable of supplying a first culture medium as the liquid to the first pool via the first supply port, and a second culture medium supply unit capable of supplying the second culture medium, the second culture medium having components different from the first culture medium as the liquid, to the second pool via the second supply port, and the control device has a culture medium supply control unit that executes a culture medium supply step of controlling the first culture medium supply unit to supply the first culture medium to the first pool and controlling the second culture medium supply unit to supply the second culture medium to the second pool.

[0008] The method for operating a bioreactor according to the present disclosure is a method for operating a bioreactor comprising: an outer cylinder centered on an axis extending in the vertical direction; an inner cylinder centered on the axis inside the outer cylinder; a rotating cylinder disposed between the outer and inner cylinders, defining a first pool between itself and the inner cylinder and defining a second pool between itself and the outer cylinder, and rotatable about the axis; and a drive unit that drives the rotating cylinder to rotate about the axis, the method including a culture medium supply step of supplying a first culture medium to the first pool and supplying a second culture medium having components different from those of the first culture medium to the second pool.

[0009] According to the present disclosure, it is possible to carry out cell culture according to the growth phase with a compact configuration.

[0010] 7 is a schematic longitudinal sectional view illustrating the overall configuration of a bioreactor system according to a first embodiment of the present disclosure. FIG. 7 is a sectional view taken along II-II of FIG. 1. FIG. 7 is a functional block diagram of a control device of a bioreactor system according to a first embodiment of the present disclosure. FIG. 7 is a flowchart illustrating a processing flow of a control device of a bioreactor system according to a first embodiment of the present disclosure. FIG. 7 is a flowchart illustrating a procedure for a culture step according to a first embodiment of the present disclosure. FIG. 7 is a flowchart illustrating a procedure for a movement step according to a first embodiment of the present disclosure. FIG. 7 is a schematic longitudinal sectional view taken along VIII-VIII of FIG. 7. FIG. 7 is a horizontal sectional view of a rotating cylinder of a bioreactor system according to a second embodiment of the present disclosure, illustrating a state in which the rotating cylinder is driven in forward rotation. FIG. 7 is a horizontal sectional view of a rotating cylinder of a bioreactor system according to a second embodiment of the present disclosure, illustrating a state in which the rotating cylinder is driven in reverse rotation. FIG. 7 is a flowchart illustrating a procedure for a movement step according to a second embodiment of the present disclosure. FIG. 7 is a horizontal sectional view of a rotating cylinder of a bioreactor system according to a modified example of the second embodiment of the present disclosure, illustrating a state in which the rotating cylinder is driven in forward rotation. FIG. 7 is a horizontal sectional view of a rotating cylinder of a bioreactor system according to a modified example of the second embodiment of the present disclosure, illustrating a state in which the rotating cylinder is driven in reverse rotation. FIG. 1 is a schematic longitudinal sectional view illustrating the overall configuration of a bioreactor system according to a third embodiment of the present disclosure. FIG. 2 is a functional block diagram of a control device of a bioreactor system according to the third embodiment of the present disclosure. FIG. 3 is a flowchart showing the processing flow of the control device of the bioreactor system according to the third embodiment of the present disclosure. FIG. 4 is a schematic longitudinal sectional view illustrating the overall configuration of a bioreactor system according to a fourth embodiment of the present disclosure. FIG. 5 is an enlarged view of a main part illustrating the configuration of a seal unit of a bioreactor system according to a fifth embodiment of the present disclosure. FIG. 6 is an enlarged view of a main part illustrating the configuration of a seal unit of a bioreactor system according to a first modified example of the fifth embodiment of the present disclosure. FIG. 7 is an enlarged view of a main part illustrating the configuration of a seal unit of a bioreactor system according to a second modified example of the fifth embodiment of the present disclosure. FIG. 8 is an enlarged view of a main part illustrating the configuration of a low thermal conductivity member of a bioreactor system according to a sixth embodiment of the present disclosure. FIG. 9 is a hardware configuration diagram of a control device of a bioreactor system according to the present disclosure.

[0011] <First Embodiment> <Bioreactor System> A first embodiment of the present disclosure will be described in detail below with reference to Figures 1 to 6. The bioreactor system 1 shown in Figure 1 is a system for performing cell culture in two stages, namely, cell growth and antibody production. The bioreactor system 1 produces antibodies as a final product. The bioreactor system 1 includes a bioreactor 10 and a control device 200.

[0012] <Bioreactor> As shown in FIG. 1 , the bioreactor 10 includes a casing 20, an inner cylinder 30, a rotating unit 40, a driving unit 50, a first culture medium supply unit 60, a second culture medium supply unit 70, a cell supply unit 80, a first temperature adjustment unit 91, a second temperature adjustment unit 92, an oxygen supply unit 100, a removal unit 110, and a moving mechanism 150.

[0013] <Casing> The casing 20 forms the outer shell of the bioreactor 10. The casing 20 has a structure centered on an axis O extending in the vertical direction. The casing 20 has a lower bottom 21, an outer cylinder 22, and a lid 23. The lower bottom 21 is a plate-like member that extends horizontally and is placed on the floor surface. In a plan view, the lower bottom 21 has a circular shape, for example, centered on the axis O.

[0014] <Outer Cylinder> The outer cylinder 22 is a cylindrical member centered on the axis O. The outer peripheral surface and inner peripheral surface of the outer cylinder 22 form cylindrical surfaces extending in the up-down direction centered on the axis O. The outer diameter and inner diameter of the outer cylinder 22 are constant in the up-down direction. The outer cylinder 22 is provided so as to extend upward from the outer peripheral edge of the lower base 21. In other words, the lower opening of the outer cylinder 22 is configured to be closed by the lower base 21.

[0015] The lid portion 23 closes the upper opening of the outer cylinder 22. The lid portion 23 is shaped like a cylinder with a closed top and a bottom. The lower end of the lid portion 23 is connected to the upper end of the outer cylinder 22 around the entire circumference. This defines an airtight space within the casing 20, ensuring a sterile environment. The lid portion 23 has a through portion 23a that passes through the lid portion 23 in the vertical direction. The through portion 23a extends in the vertical direction along the axis O.

[0016] <Inner cylinder> The inner cylinder 30 is a cylindrical member centered on the axis O, and is provided inside the outer cylinder 22. The outer peripheral surface of the inner cylinder 30 forms a cylindrical surface extending in the vertical direction centered on the axis O. The outer diameter of the inner cylinder 30 is set to be smaller than the inner diameter of the outer cylinder 22 and is constant in the vertical direction. The lower end of the inner cylinder 30 forms a circular flat surface that extends in the horizontal direction. The entire lower end of the inner cylinder 30 is fixed integrally to the lower bottom 21. The upper end of the inner cylinder 30 forms a circular flat surface that extends in the horizontal direction. The height of the upper end of the inner cylinder 30 is set to be a height that can be accommodated in the space within the casing 20.

[0017] <Rotating Unit> The rotating unit 40 has a shaft portion 41, an upper base portion 42, and a rotating cylinder 43. The shaft portion 41 is a shaft-shaped member extending along the axis O and is installed so as to pass vertically through a through-hole in the lid portion 23. The shaft portion 41 is rotatable about the axis O, for example, via a bearing (not shown) installed in the through-hole. The upper base portion 42 is connected to the lower end of the shaft portion 41 inside the casing 20. The upper base portion 42 is a disk-shaped member whose center is the axis O and extends horizontally. The upper base portion 42 is disposed above the upper end of the inner cylinder 30 and spaced apart from the upper end. The center of the upper surface of the upper base portion 42 is fixed to the lower end of the shaft portion 41. The outer diameter of the upper base portion 42 is smaller than the inner diameter of the outer cylinder 22 and larger than the outer diameter of the inner cylinder 30.

[0018] <Rotating cylinder> The rotating cylinder 43 is a cylindrical member centered on the axis O, and is disposed between the outer cylinder 22 and the inner cylinder 30 inside the casing 20. The outer and inner peripheral surfaces of the rotating cylinder 43 form cylindrical surfaces extending in the vertical direction around the axis O. The outer diameter of the rotating cylinder 43 is set to be smaller than the inner diameter of the outer cylinder 22. The inner diameter of the rotating cylinder 43 is set to be larger than the outer diameter of the inner cylinder 30. The outer and inner diameters of the rotating cylinder 43 are constant in the vertical direction.

[0019] The upper end of the rotating cylinder 43 is integrally connected to the outer peripheral edge of the upper base 42 over the entire circumferential direction. That is, the upper opening of the inner cylinder 30 is closed by the upper base 42. The lower edge 43a of the rotating cylinder 43 faces the upper surface of the lower base 21 with a small clearance therebetween. The size of this clearance is small compared to the overall size of the bioreactor 10. The vertical dimension of the clearance is set to, for example, 1 / 1000 to 1 / 100 of the vertical length of the casing 20. The dimension of the clearance is set to, for example, 1 mm to 10 mm.

[0020] The rotating cylinder 43 is rotatable around the axis O together with the shaft 41 and the upper base 42. A clearance is formed between the lower end of the inner cylinder 30 and the lower base plate, allowing the rotating cylinder 43 to rotate smoothly without interfering with the lower base 21. Note that the lower end of the rotating cylinder 43 may be configured to slidably contact the upper surface of the upper base 42.

[0021] <First Pool, Second Pool> The rotating cylinder 43 defines two independent spaces, a first pool P1 and a second pool P2, within the casing 20. The first pool P1 and the second pool P2 are each capable of containing a liquid. The contained liquid is a culture solution containing a culture medium and cells. The first pool P1 is defined by the outer peripheral surface of the inner cylinder 30 and the inner peripheral surface of the rotating cylinder 43. The first pool P1 is an annular space centered on the axis O and extending vertically. The second pool P2 is defined by the outer peripheral surface of the rotating cylinder 43 and the inner peripheral surface of the outer cylinder 22. The second pool P2 is an annular space centered on the axis O and extending vertically, and is one size larger than the first pool P1. The first pool P1 and the second pool P2 are slightly connected via the clearance described above. Because the clearance is small, there is almost no movement of culture solution between the first pool P1 and the second pool P2.

[0022] <Drive unit> The drive unit 50 drives the rotating unit 40 to rotate around the axis O. The drive unit 50 is provided outside the casing 20. The drive unit 50 is, for example, an electric motor, and its output shaft is connected to the upper end of the shaft 41 of the rotating unit 40. When the output shaft of the drive unit 50 rotates at a predetermined rotation speed, the rotating unit 40 is driven to rotate at the same rotation speed.

[0023] The driving unit 50 may be provided inside the casing 20. In this case, for example, the driving unit 50 may be installed at the upper end of the inner cylinder 30, and the rotating unit 40 may be rotationally driven by the driving unit 50.

[0024] <First Culture Medium Supply Unit> The first culture medium supply unit 60 supplies the first culture medium to the first pool P1. The first culture medium supply unit 60 has a first supply pipe 61, a first supply line 62, a first supply valve 63, and a first culture medium storage unit 64. The first supply pipe 61 is a pipe inserted from the outside of the casing 20 and has a first supply port 61a that opens toward the first pool P1. The first supply pipe 61 in this embodiment penetrates the inner cylinder 30, and is provided so that the first supply port 61a opens to the bottom of the first pool P1.

[0025] The first supply line 62 is a pipe extending outside the casing 20, and is connected to the end of the first supply pipe 61 opposite to the first supply port 61 a. The first supply valve 63 is provided on the first supply line 62, and is configured to be able to open and close the flow path of the first supply line 62.

[0026] The first culture medium storage section 64 stores a first culture medium. The first culture medium is adjusted to have components suitable for cell growth. The first culture medium storage section 64 is connected to the end of the first supply line 62 opposite the first culture medium supply pipe. The first culture medium supply section 60 is configured such that, by switching the first supply valve 63 from a closed state to an open state, the first culture medium in the first culture medium storage section 64 is supplied into the first pool P1 via the first supply line 62 and the first supply pipe 61 and the first supply port 61 a.

[0027] <Second Culture Medium Supply Unit> The second culture medium supply unit 70 supplies the second culture medium to the second pool P2. The second culture medium supply unit 70 has a second supply pipe 71, a second supply line 72, a second supply valve 73, and a second culture medium storage unit 74. The second supply pipe 71 is a pipe inserted from the outside of the casing 20 and has a second supply port 71a that opens toward the second pool P2. The second supply pipe 71 in this embodiment is provided above the second pool P2 so that the second supply port 71a opens downward.

[0028] The second supply line 72 is a pipe extending outside the casing 20, and is connected to the end of the second supply pipe 71 opposite to the second supply port 71 a. The second supply valve 73 is provided on the second supply line 72, and is configured to be able to open and close the flow path of the second supply line 72.

[0029] The second culture medium storage section 74 stores a second culture medium. The second culture medium is adjusted to have components suitable for cells to produce antibodies while suppressing cell proliferation. The second culture medium storage section 74 is connected to the end of the second supply line 72 opposite the second culture medium supply pipe. The second culture medium supply section 70 is configured such that, by switching the second supply valve 73 from a closed state to an open state, the second culture medium in the second culture medium storage section 74 is supplied into the second pool P2 via the second supply line 72 and the second supply pipe 71 and the second supply port 71 a.

[0030] <Cell Supply Unit> The cell supply unit 80 supplies cells to the first pool P1. The cell supply unit 80 has a cell supply pipe 81, a cell supply line 82, a cell supply valve 83, and a cell storage unit 84. The cell supply pipe 81 is a pipe inserted from the outside of the casing 20 and has a cell supply port 81a that opens toward the cell pool. In this embodiment, the cell supply pipe 81 penetrates the inner cylinder 30, and the cell supply port 81a is provided so as to open to the bottom of the first pool P1.

[0031] The cell supply line 82 is a pipe extending outside the casing 20, and is connected to the end opposite the cell supply port 81a of the cell supply pipe 81. The cell supply valve 83 is provided on the cell supply line 82, and is configured to be able to open and close the flow path of the cell supply line 82.

[0032] The cell storage unit 84 stores cells to be cultured. The cell storage unit 84 is connected to the end of the cell supply line 82 opposite to the cell supply pipe 81. The cell supply unit 80 is configured such that, by switching the cell supply valve 83 from a closed state to an open state, the cells in the cell storage unit 84 are supplied into the cell pool via the cell supply line 82 and the cell supply pipe 81 and the cell supply port 81 a.

[0033] <First Temperature Adjustment Unit> The first temperature adjustment unit 91 adjusts the temperature of the first pool P1. The first temperature adjustment unit 91 sets the temperature of the first pool P1 to a predetermined temperature by raising or lowering the temperature of the first pool P1. The first temperature adjustment unit 91 is provided inside the inner cylinder 30. That is, the first temperature adjustment unit 91 is provided inside the inner cylinder 30 in the circumferential direction and the up-down direction, and is capable of adjusting the temperature of the first pool P1 via the outer peripheral surface of the inner cylinder 30.

[0034] <Second Temperature Adjustment Unit> The second temperature adjustment unit 92 adjusts the temperature of the second pool P2. The second temperature adjustment unit 92 sets the temperature of the second pool P2 to a predetermined temperature by raising or lowering the temperature of the second pool P2. The second temperature adjustment unit 92 is provided to surround the outer periphery of the outer cylinder 22. The second temperature adjustment unit 92 is annular and is provided to extend circumferentially and vertically around the outer cylinder 22. In other words, the second temperature adjustment unit 92 is capable of adjusting the temperature of the first pool P1 via the inner circumferential surface of the outer cylinder 22.

[0035] As the first temperature adjustment unit 91 and the second temperature adjustment unit 92, various temperature adjustment mechanisms such as a heater, a cooler, a Peltier element, etc. can be adopted.

[0036] <Oxygen Supply Unit> The oxygen supply unit 100 supplies oxygen to the first pool P1 and the second pool P2. As shown in FIGS. 1 and 2 , the oxygen supply unit 100 of this embodiment is disposed on the upper surface of the lower bottom 21. A plurality of oxygen supply units 100 are provided at intervals in the circumferential direction. Each oxygen supply unit 100 extends in the radial direction of the axis O and is provided so as to straddle the first pool P1 and the second pool P2 across the rotating cylinder 43 in plan view. Each oxygen supply unit 100 is a sparger capable of releasing oxygen. The oxygen concentration of the culture solution in the first pool P1 and the second pool P2 is adjusted by releasing oxygen from the sparger.

[0037] <Extraction Unit> The extraction unit 110 has a role of extracting the liquid in the second pool P2 to the outside. The extraction unit 110 has a discharge line 111, a discharge valve 112, an extraction pump 113, a cell acquisition unit 120, and an antibody acquisition unit 130. The discharge line 111 is a pipe provided outside the casing 20, and one end is connected to the second pool P2. The discharge valve 112 is provided on the discharge line 111 and can be opened and closed. The extraction pump 113 is provided on the discharge line 111 and can pump the fluid from one end of the discharge line 111 to the other end.

[0038] The cell acquisition unit 120 is a filter capable of extracting cells from the culture solution. The cell acquisition unit 120 is provided upstream of the discharge valve 112 and the extraction pump 113 in the discharge line 111. The antibody acquisition unit 130 is a filter capable of extracting antibodies produced by cells from the culture solution. The antibody acquisition unit 130 is provided upstream of the discharge valve 112 and the extraction pump 113 in the discharge line 111 and downstream of the cell acquisition unit 120.

[0039] With the above-described configuration, the take-out section 110 is capable of discharging the culture solution in the second pool P2 to the outside by closing the discharge valve 112 and operating the take-out pump 113.

[0040] <Movement Mechanism> The movement mechanism 150 is a mechanism capable of moving the liquid in the first pool P1 to the second pool P2. The movement mechanism 150 of this embodiment has a movement line 151 and a movement valve 152. The movement line 151 is a pipe arranged outside the casing 20, and one end is connected to the first pool P1. The other end of the movement line 151 is connected to the second pool P2. In this embodiment, the other end of the movement line 151 is connected to the discharge line 111 on the upstream side of the cell acquisition unit 120. As a result, the movement line 151 is connected to the second pool P2 via the discharge line 111. Note that the other end of the movement line 151 may be configured to be directly connected to the second pool P2.

[0041] The transfer valve 152 is provided on the discharge line 111 and can be opened and closed. When the liquid level in the second pool P2 is lower than the liquid level in the first pool P1, the transfer valve 152 is switched from a closed state to an open state, and the liquid in the first pool P1 moves to the second pool P2 due to the head difference.

[0042] <Control Device> Next, the control device 200 of this embodiment will be described with reference to Fig. 3. As shown in Fig. 3, the control device 200 includes a culture medium supply control unit 201, a temperature setting unit 202, a cell supply control unit 203, a culture execution unit 204, a movement execution unit 205, a discharge execution unit 206, a valve control unit 211, an oxygen supply control unit 212, a rotation control unit 213, and a pump control unit 214.

[0043] The culture medium supply control unit 201 controls the first culture medium supply unit 60 and the second culture medium supply unit 70 via the valve control unit 211 to supply the first culture medium to the first pool P1 and the second culture medium to the second pool P2. The temperature setting unit 202 controls the first temperature adjustment unit 91 and the second temperature adjustment unit 92 so that the first pool P1 and the second pool P2 reach their target temperatures, respectively. The cell supply control unit 203 controls the cell supply unit 80 via the valve control unit 211 to supply the first culture medium to the first pool P1.

[0044] The culture execution unit 204 rotates the rotation unit 40 by controlling the drive unit 50 via the rotation control unit 213. The culture execution unit 204 supplies oxygen into the first pool P1 and the second pool P2 by controlling the oxygen supply unit 100 via the oxygen supply control unit 212. The transfer execution unit 205 transfers the culture medium from the first pool P1 to the second pool P2 by controlling the transfer mechanism 150 via the valve control unit 211. The discharge execution unit 206 controls the take-out unit 110 via the valve control unit 211 and the pump control unit 214 to take out the culture medium from the second pool P2.

[0045] The valve control unit 211 controls the opening and closing operations of the first supply valve 63, the second supply valve 73, the cell supply valve 83, the movement valve 152, and the discharge valve 112 in response to instructions from the culture medium supply control unit 201, the cell supply control unit 203, the movement execution unit 205, and the discharge execution unit 206.

[0046] The oxygen supply control unit 212 controls the oxygen supply unit 100 in response to a command from the culture execution unit 204. The rotation control unit 213 controls the drive unit 50 to rotate the rotation unit 40 in response to a signal from the culture execution unit 204. The pump control unit 214 controls the extraction pump 113 in response to a command from the discharge execution unit 206.

[0047] <Method of Operating Bioreactor> Next, the processing procedure of the control device 200 (method of operating the bioreactor 10) will be described with reference to the flowchart shown in Figure 4. The control device 200 sequentially performs a medium supplying step S1, a temperature setting step S2, a cell supplying step S3, a culturing step S4, a moving step S5, a culturing step S6, and a discharging step S7.

[0048] <Culture Medium Supply Step> First, the culture medium supply control unit 201 outputs a signal to the valve control unit 211, which causes the valve control unit 211 to change the first supply valve 63 and the second supply valve 73 from a closed state to an open state. As a result, the first culture medium in the first culture medium storage unit 64 is supplied to the first pool P1 via the first supply port 61a of the first supply pipe 61. Furthermore, the second culture medium in the second culture medium storage unit 74 is supplied to the second pool P2 via the second supply port 71a of the second supply pipe 71 (culture medium supply step S1). Once appropriate amounts of the first culture medium and the second culture medium have been supplied to the first pool P1 and the second pool P2, the valve control unit 211 changes the first supply valve 63 and the second supply valve 73 from an open state to a closed state. This stops the supply of the first culture medium and the second culture medium. Note that, to smoothly perform the transfer step described below, the first culture medium supply unit 60 and the second culture medium supply unit 70 may be controlled so that the liquid level of the first culture medium is higher than the liquid level of the second culture medium.

[0049] <Temperature Setting Step> Next, the temperature setting unit 202 controls the first temperature adjustment unit 91 and the second temperature adjustment unit 92 to adjust the first medium in the first pool P1 and the second medium in the second pool P2 to a predetermined temperature (temperature setting step S2). Here, the temperature of the first medium is set to a temperature suitable for cell growth. That is, the first pool P1 is set to a temperature of, for example, 36 to 38°C, preferably around 37°C. Meanwhile, the temperature of the second medium is set to a temperature that suppresses cell growth while also suitable for antibody production by the cells. That is, the second pool P2 is set to a lower temperature than the first pool P1, for example, 30 to 33°C, preferably 31 to 32°C.

[0050] <Cell Supply Step> Thereafter, the cell supply control unit 203 outputs a signal to the valve control unit 211, which causes the valve control unit 211 to change the cell supply valve 83 from a closed state to an open state. As a result, the cells in the cell storage unit 84 are supplied to the first pool P1 via the cell supply port 81a of the cell supply pipe 81 (cell supply step S3). When an appropriate amount of cells has been supplied to the first pool P1, the valve control unit 211 changes the cell supply valve 83 from an open state to a closed state. As a result, the supply of cells is stopped.

[0051] <Culturing Step> Next, the culturing execution unit 204 cultures cells in the first pool P1 (culturing step S4). The culturing step S4 is performed according to the procedure shown in the flowchart in Fig. 5. That is, first, the culturing execution unit 204 outputs a signal to the oxygen supply control unit 212, which then activates the oxygen supply unit 100. This starts the supply of oxygen to the first pool P1 and the second pool P2 (step S41).

[0052] Next, the culture execution unit 204 sends a signal to the rotation control unit 213, which controls the drive unit 50 to rotate the rotation unit 40 (step S42). Here, the rotation speed of the rotation unit 40 is set to a value suitable for forming Taylor vortices in the first pool P1 and the second pool P2. The rotation control unit 213 drives the rotation cylinder 43 in the forward direction so that the Taylor number in the first pool P1 and the second pool P2 is, for example, 20,000 to 30,000, taking into account the radial dimensions of the first pool P1 and the second pool P2. This makes it difficult for turbulence to occur in the first pool P1 and the second pool P2, resulting in agitation by a stable vortex, promoting cell proliferation in the first pool P1.

[0053] Next, the culture execution unit 204 determines whether the culture is complete (step S43). For example, when the cell concentration in the first pool P1 measured by a cell counter does not exceed a predetermined value, the culture execution unit 204 determines that the culture is not complete and continues driving the rotating cylinder 43 in the forward direction (step S43: No). For example, when the cell concentration does not exceed a predetermined value, the culture execution unit 204 determines that the culture is not complete (step S43: Yes). Note that the culture execution unit 204 may determine that the culture is complete when a predetermined time has elapsed after driving the rotating cylinder 43 in rotation.

[0054] After the culture is completed, the culture execution unit 204 outputs a signal to the oxygen supply control unit 212, which causes the oxygen supply control unit 212 to stop the supply of oxygen by the oxygen supply unit 100 (step S45). Thereafter, the culture execution unit 204 outputs a signal to the rotation control unit 213, which causes the rotation control unit 213 to stop the rotation of the rotating cylinder 43 (step S46). This completes the culture step S4, and the first culture medium becomes filled with the proliferated cells.

[0055] <Transfer Step> Next, as shown in FIG. 4 , after the culture step S4, the transfer execution unit 205 transfers the culture medium from the first pool P1 to the second pool P2 (transfer step S5). As a result, the cells grown in the first pool P1 are supplied to the second culture medium in the second pool P2. As shown in FIG. 6 in more detail, in the transfer step S5, the transfer execution unit 205 outputs a signal to the valve control unit 211, which causes the valve control unit 211 to open the transfer valve 152 (S51). As a result, if the liquid level in the first pool P1 is higher than the liquid level in the second pool P2, the head difference causes the culture medium in the first pool P1 to transfer to the second pool P2 via the transfer line 151.

[0056] If the liquid level in the first pool P1 is not higher than the liquid level in the second pool P2, the transfer execution unit 205 may control the valve control unit 211 to temporarily open the discharge valve 112 and control the pump control unit 214 to temporarily operate the extraction pump 113 prior to opening the transfer valve 152. This allows the liquid in the second pool P2 to be discharged to the outside, thereby making the liquid level in the first pool P1 higher than the liquid level in the second pool P2. Note that a pump may be provided in the transfer line 151, and the culture medium may be moved from the first pool P1 to the second pool P2 by operating the pump.

[0057] Next, the movement execution unit 205 determines whether the movement of the culture medium is complete (step S52). For example, if a predetermined time has not elapsed since the movement valve was opened, the movement execution unit 205 determines that the movement is not complete (step S52: No). On the other hand, if a predetermined time has elapsed since the movement valve was opened, the movement execution unit 205 determines that the movement is complete (step S52: Yes). Note that the movement execution unit 205 may determine that the movement of the culture medium is complete when the liquid levels in the first pool P1 and the second pool P2 are equal. Alternatively, the cell concentrations in the first pool P1 and the second pool P2 may be detected, and the movement of the culture medium may be determined to be complete based on the detected cell concentrations.

[0058] After the transfer of the culture medium is completed, the transfer execution unit 205 outputs a signal to the valve control unit 211, which then changes the transfer valve 152 from the open state to the closed state (step S53). This completes the transfer step S5.

[0059] <Repeated Cultivation Step After Transfer Step> After completion of the transfer step S5, a cultivating step S6 similar to the cultivating step S4 is performed as shown in FIG. 4. This agitates the culture medium in the second pool P2, promoting antibody production by the cells in the second pool P2. In the cultivating step S6 after the transfer step S5, the culture is determined to be complete when the detected cell concentration in the second pool P2 begins to decrease, indicating that antibody production has ended (FIG. 5: step S43). Alternatively, the culture may be determined to be complete when a predetermined time has elapsed.

[0060] <Discharge Step> After the completion of the second culture step S6 following the transfer step S5, the culture medium in the second pool P2 is discharged by the discharge execution unit 206 (discharge step S7). Specifically, the discharge execution unit 206 controls the valve control unit 211 to change the discharge valve 112 from a closed state to an open state, and controls the pump control unit 214 to operate the extraction pump 113. As a result, the culture medium in the second pool P2, together with the cells and antibodies, is discharged from the second pool P2 via the discharge line 111. During the process of flowing through the discharge line 111, the cells in the culture medium are extracted by the cell acquisition unit 120, and the antibodies in the culture medium are extracted by the antibody acquisition unit 130. The extracted cells can be reused by supplying them again to the first pool P1. The extracted antibodies can be used for various purposes as products of this system. This completes the operating method of the bioreactor 10.

[0061] <Effects> According to the bioreactor 10, bioreactor system 1, and method of operating the bioreactor 10 described above, cell culture can be performed in the first pool P1 and the second pool P2, which are independent of each other. Therefore, cells in different growth phases, such as cell proliferation and antibody production, can be cultured in a single device. This eliminates the need to use multiple devices, and the overall system can be made more compact.

[0062] Furthermore, since the first pool P1 and the second pool P2 can be supplied with media containing different components, it is possible to supply media containing components suitable for cell proliferation and antibody production, respectively, thereby enabling efficient cell culture in each growth phase.

[0063] To stir the culture medium in each of the first and second pools P1 and P2, it is possible to rotate the inner cylinder 30 and the outer cylinder 22, for example. However, this would complicate the device configuration because two cylinders must be rotated. Furthermore, rotating the outer cylinder 22 requires more power than rotating the rotating cylinder 43. In this embodiment, the first and second pools P1 and P2 can be stirred by rotating only the rotating cylinder 43, resulting in a simple and efficient device configuration. Furthermore, since the inner cylinder 30 does not need to be rotated in this embodiment, the first culture medium supply pipe and the cell supply pipe 81 can be installed so as to pass through the inner cylinder 30. This allows for efficient use of the internal space of the inner cylinder 30, thereby reducing the overall installation space of the device.

[0064] Furthermore, the temperatures of the first pool P1 and the second pool P2 are controlled by the first temperature control unit 91 and the second temperature control unit 92, which can set temperatures independently, so that the first pool P1 and the second pool P2 can be set to temperatures suitable for their respective cultures. This allows for more efficient cell culture in different growth phases.

[0065] Furthermore, antibody production can be carried out by transferring the cells grown in the first pool P1 to the second pool P2. In other words, a series of processes from cell growth to antibody production can be completed in a single device, allowing for efficient antibody production.

[0066] Furthermore, in this embodiment, the oxygen supply unit 100 is disposed across the first pool P1 and the second pool P2, so that oxygen can be efficiently supplied to both the first pool P1 and the second pool P2, thereby creating an environment suitable for the cells.

[0067] Second Embodiment Next, a second embodiment of the present disclosure will be described with reference to Figures 7 to 13. In Figures 7 to 13, components similar to those in the first embodiment are denoted by the same reference numerals, and detailed description thereof will be omitted. The bioreactor system 1 of the second embodiment differs from the first embodiment in the configurations of the drive unit 50, the oxygen supply unit 100, and the movement mechanism 150.

[0068] <Drive Unit> The drive unit 50 is capable of driving the rotation unit 40 to rotate in both the forward and reverse directions. That is, the drive unit 50 drives the rotation cylinder 43 to rotate forward and backward. Note that in the culture steps S4 and S6, the culture execution unit 204 controls the drive unit 50 via the rotation control unit 213 to rotate the rotation cylinder 43 forward. <Oxygen Supply Unit> As shown in FIGS. 7 and 8 , the oxygen supply unit 100 of the second embodiment includes a first oxygen supply unit 101 and a second oxygen supply unit 102. The first oxygen supply unit 101 and the second oxygen supply unit 102 are spargers capable of supplying oxygen into the liquid. The first oxygen supply unit 101 is disposed in a region between the inner cylinder 30 and the rotation cylinder 43 on the upper surface of the lower bottom 21 of the casing 20. That is, the first oxygen supply unit 101 is disposed so as to face only the first pool P1 out of the first pool P1 and the second pool P2.

[0069] The second oxygen supply unit 102 is disposed in a region between the outer cylinder 22 and the rotating cylinder 43 on the upper surface of the lower bottom portion 21 of the casing 20. That is, the second oxygen supply unit 102 is disposed so as to face only the second pool P2 out of the first pool P1 and the second pool P2.

[0070] The first oxygen supply unit 101 and the second oxygen supply unit 102 are independent of each other and each releases a set supply amount of oxygen. That is, in the culture steps S4 and S6, the oxygen supply control unit 212 controls the first oxygen supply unit 101 and the second oxygen supply unit 102 so that the oxygen supply amount is appropriate for the first pool P1 and the second pool P2, which are the oxygen supply destinations.

[0071] As a result, the first oxygen supply unit 101 supplies an amount of oxygen suitable for cell proliferation, for example, to promote cell proliferation in the first pool P1, while the second oxygen supply unit 102 supplies an amount of oxygen suitable for antibody production, for example, to suppress cell proliferation in the second pool P2 and promote antibody production by the cells.

[0072] <Moving Mechanism> The moving mechanism 150 of the second embodiment is provided on the rotating cylinder 43. As shown in Figures 7, 9 and 10, a plurality of openings 43b that penetrate the rotating cylinder 43 in the radial direction are provided at the bottom of the rotating cylinder 43 at intervals in the circumferential direction.

[0073] Furthermore, the movement mechanism 150 has a sliding door 160 as an opening / closing part that can open and close the opening 43b. The sliding door 160 has a plate shape that extends circumferentially and vertically, and is sized to close the opening 43b. The sliding door 160 is provided along the outer peripheral surface of the rotating cylinder 43. The sliding door 160 is provided so as to be movable between a closed position that closes the opening 43b from the outer peripheral side, and an open position that opens the opening 43b. The open position is a position closer to the front in the rotational direction of the rotating cylinder 43 than the closed position. The sliding door 160 is provided so as to be slidable in the circumferential direction along the outer peripheral surface of the rotating cylinder 43 between the closed position and the open position.

[0074] The movement mechanism 150 has a stopper 161. The stopper 161 is provided so as to protrude outward from the rear edge of the opening 43b of the rotating cylinder 43 in the forward rotation direction. When the sliding door 160 is in the closed position, it abuts against the stopper 161 from the front side in the forward rotation direction. The stopper 161 restricts the movement of the sliding door 160 from the closed position toward the rear side in the forward rotation direction.

[0075] Next, the movement step S5 of this embodiment will be described with reference to FIG. 11 . First, the movement execution unit 205 determines whether a predetermined time has elapsed after the completion of the initial incubation step S4, i.e., after the rotation of the rotating cylinder 43 has stopped (step S54). If the predetermined time has not elapsed, the movement execution unit 205 waits until the predetermined time has elapsed (step S53: No). If the predetermined time has elapsed, the movement execution unit 205 proceeds to the next step (step S53: Yes). In this way, by waiting after the rotation of the rotating cylinder 43 has stopped, the cells that have proliferated in the culture medium in the first pool P1 settle to the bottom of the first pool P1. This results in a higher cell concentration in the bottom of the first pool P1 (the vertical position of the opening / closing unit) compared to the top.

[0076] Next, the movement execution unit 205 drives the drive unit 50 in reverse rotation via the rotation control unit 213. Accordingly, the rotating cylinder 43 rotates in the reverse direction opposite to that during the incubation step S4 (step S55). Here, during forward rotation of the rotating cylinder 43, the flow resistance of the liquids in the first pool P1 and the second pool P2 is applied to the sliding door 160, and an external force is applied to the sliding door 160 toward the rear in the forward rotation direction. This causes the sliding door 160 to remain in contact with the stopper 161 in the closed position. Therefore, during forward rotation of the rotating cylinder 43, the sliding door 160 closes the opening 43b, preventing the liquids in the first pool P1 and the second pool P2 from mixing through the opening 43b.

[0077] On the other hand, when the rotating cylinder 43 is rotated in the reverse direction in the movement step S5, an external force is applied to the sliding door 160 toward the rear in the reverse rotation direction (forward in the forward rotation direction) due to the flow resistance of the liquid in the first pool P1 and the second pool P2. As a result, the sliding door 160 moves toward the rear in the reverse rotation direction relative to the rotating cylinder 43. That is, the sliding door 160 slides from the closed position to the open position. As a result, the first pool P1 and the second pool P2 are radially connected via the sliding door 160 of the rotating cylinder 43.

[0078] Here, the movement execution unit 205 controls the rotation control unit 213 to rotate the rotating cylinder 43 in the reverse direction at a lower rotation speed than during the culture steps S4 and S6. As a result, Taylor vortices are not formed in the first pool P1 and the second pool P2, and active mixing is not performed. Meanwhile, as the rotating cylinder 43 comes into contact with the culture solution in the first pool P1 at a low rotation speed, the culture solution in the first pool P1 rotates along with the rotating cylinder 43 due to viscosity. This applies a gentle centrifugal force to the culture solution in the first pool P1. This centrifugal force causes the culture solution with a high cell concentration in the first pool P1 to move to the second pool P2 through the opening 43b.

[0079] Next, the movement execution unit 205 determines whether the movement of the culture medium from the first pool P1 to the second pool P2 is complete (step S56). If it is determined that the movement is not complete (step 56: No), the movement execution unit 205 continues the reverse rotation of the rotating cylinder 43. If it is determined that the movement is complete (step 56: Yes), the movement execution unit 205 controls the rotation control unit 213 to stop the reverse rotation of the rotating cylinder 43 (step S57). This completes the movement step S5.

[0080] The determination of whether the movement by the movement execution unit 205 is complete may be made, for example, based on whether a predetermined time has elapsed since the start of reverse rotation of the rotating cylinder 43. Alternatively, the cell concentrations of the liquid in the first pool P1 and the second pool P2 may be measured, and the movement may be determined to be complete when the cell concentration in the first pool P1 falls below a predetermined level and the cell concentration in the second pool P2 exceeds the predetermined level. Alternatively, the cell concentration in either the first pool P1 or the second pool P2 may be used as the determination criterion.

[0081] <Effects> In the bioreactor system 1 of the second embodiment configured as described above, oxygen can be supplied separately to each of the first pool P1 and the second pool P2. Therefore, when cells in different growth phases are present in the first pool P1 and the second pool P2, an appropriate amount of oxygen can be supplied to each cell, thereby promoting culture. In particular, in this embodiment, an amount of oxygen appropriate for cell culture is supplied to the first pool P1, and an amount of oxygen appropriate for antibody production is supplied to the second pool P2, thereby enabling the two processes of cell culture and antibody production to be carried out efficiently.

[0082] Furthermore, according to the movement mechanism 150 of this embodiment, the culture medium can be moved from the first pool P1 to the second pool P2 simply by the reverse rotation of the rotary cylinder 43. Therefore, the movement of the culture medium can be performed smoothly and appropriately. Furthermore, the only objects to be controlled are the drive unit 50 and the rotary cylinder 43, and the movement valve 152 of the first embodiment is not required. Therefore, the number of parts is reduced, thereby improving maintainability.

[0083] <Modification of Second Embodiment> As a modification of the second embodiment, the movement mechanism 150 may have the configuration shown in Figures 12 and 13, for example. The modification movement mechanism 150 has an open / close door 170 and a hinge 171 instead of the sliding door 160 and stopper 161 of the second embodiment. The hinge 171 is provided on the inner circumferential surface of the rotating cylinder 43 at the front edge in the forward rotation direction of the opening 43b so as to extend in the vertical direction.

[0084] The door 170 is provided inside the rotating cylinder 43 by a hinge 171 so as to be rotatable about a rotation axis extending in the vertical direction. The door 170 rotates between a closed position and an open position. As shown in FIG. 12 , the door 170 is positioned so as to follow the shape of the cylindrical surface and closes the opening 43 b. As shown in FIG. 13 , the door 170 is positioned so as to rotate from the closed position toward the inside of the rotating cylinder 43 and open the opening 43 b.

[0085] During forward rotation of the rotating cylinder 43, the flow resistance of the first pool P1 is applied to the door 170, and an external force is applied to the door 170 toward the rear in the forward rotation direction, i.e., toward the front in the reverse rotation direction. This causes the door 170 to remain in the closed position, and the opening 43b is closed. On the other hand, when the rotating cylinder 43 is rotated in the reverse direction, the flow resistance of the first pool P1 applies an external force to the door 170, moving it from the closed position to the open position. This opens the opening 43b, and the first pool P1 and the second pool P2 become connected. Therefore, as in the second embodiment, the culture medium can be moved from the first pool P1 to the second pool P2.

[0086] <Third Embodiment> Next, a bioreactor system 1 according to a third embodiment will be described with reference to Figures 14 to 16. In Figures 14 to 16, components similar to those in the second embodiment are denoted by the same reference numerals, and detailed description thereof will be omitted. In the third embodiment, the discharge unit 110 further includes a circulation line 140 and a circulation valve 141. <Circulation Line> One end of the circulation line 140 is connected to a portion of the discharge line 111 between the discharge pump 113 and the discharge valve 112. The other end of the circulation line 140 is connected to the inside of the second pool P2. The other end of the circulation line 140 is arranged to face the culture solution in the second pool P2 from above.

[0087] <Circulation Valve> The circulation valve 141 is provided on the circulation line 140. The circulation valve 141 switches the circulation line 140 between a flowing state and a non-flowing state by opening and closing.

[0088] <Control Device> As shown in Fig. 15, the control device 200 of the third embodiment has a circulation execution unit 207 and a determination unit 208 in addition to the configuration of the control device 200 of the first embodiment shown in Fig. 3. The circulation execution unit 207 controls the opening and closing of the circulation valve 141 via a valve control unit 211, and controls the extraction pump 113 via a pump control unit 214. The determination unit 208 determines whether or not to perform continuous operation.

[0089] Next, the processing procedure of the control device 200 of the third embodiment (the method of operating the bioreactor 10) will be described with reference to the flowchart shown in Figure 16. First, as in the first embodiment, the medium supplying step S11, the temperature setting step S12, the cell supplying step S13, the culturing step S14, and the transfer step S15 are performed in this order. In the configuration of the bioreactor system 1 of the third embodiment shown in Figure 14, the transfer step S15 is performed in the same manner as in the second embodiment, but may also be performed in the same manner as in the first embodiment.

[0090] After the transfer step S15, the cell supply step S16 is performed again in the same manner as described above. Here, by performing the transfer step S15, most of the cells in the first pool P1 have been transferred to the second pool P2. Therefore, the first pool P1 is in a state where the number of cells is reduced. By performing the cell supply step S16, cells are present in both the first pool P1 and the second pool P2. Note that if the number of cells in the first pool P1 is secured after the transfer step S15, the cell supply step S16 may be omitted.

[0091] Next, the culture execution unit 204 executes the culture step S17 again, similar to the above. As a result, Taylor vortices are formed in the first pool P1 and the second pool P2 due to the rotation of the rotating cylinder 43. In this embodiment, because cells exist in both the first pool P1 and the second pool P2, cell culture proceeds in both the first pool P1 and the second pool P2. That is, cell proliferation proceeds in the first pool P1, and antibody production proceeds in the second pool P2.

[0092] After the culturing step S17 following the transferring step S15, the determining unit 208 determines whether or not to continue operation (determining step S18).

[0093] The determination of whether to perform continuous operation is made based on, for example, whether the required amount of antibody production has been reached. That is, if the discharge step S20 is executed in the next step, and it is not expected that the target amount of antibody production will be reached by obtaining antibodies in the discharge step S20, the determination unit 208 determines to perform continuous operation. On the other hand, if it is expected that the target amount of antibody production will be reached, the determination unit 208 determines not to perform continuous operation. Note that the determination unit 208 may determine whether to perform continuous operation based on the number of times it has been determined that continuous operation should be performed.

[0094] If the determination unit 208 determines that continuous operation is to be performed (determination step S18: Yes), the circulation execution unit 207 performs the circulation step S19. That is, the circulation execution unit 207 opens the circulation valve 141 via the valve control unit 211 and drives the extraction pump 113 via the pump control unit 214. Note that in this state, the discharge valve 112 is closed. As a result, the culture medium in the second pool P2 is discharged to the outside via the discharge line 111, and cells and antibodies are extracted by the cell acquisition unit 120 and the antibody acquisition unit 130. Then, the medium remaining after the cells and antibodies have been extracted from the culture medium is returned to the second pool P2 via the circulation line 140.

[0095] After the circulation step S19, the above-mentioned transfer step S15, cell supply step S16, culture step S17, and determination step S18 are performed again. Here, the cell supply step S16 may also be omitted.

[0096] If the determination unit 208 determines that continuous operation is not to be performed (determination step S18: No), the discharge step S20 is performed as in the first embodiment, whereby the final extraction of cells and antibodies is performed, and the processing by the control device 200 is completed.

[0097] As described above, according to this embodiment, cell culture can be performed simultaneously in each of the first pool P1 and the second pool P2. That is, by partitioning the first pool P1 and the second pool P2, cell culture in different growth phases, such as cell proliferation and antibody production, can be performed simultaneously. This not only improves the efficiency of cell culture, but also allows different cell cultures to be performed using only one bioreactor 10, thereby making the entire cell culture equipment more compact.

[0098] Furthermore, continuous operation allows cell growth in different growth phases, i.e., cell proliferation and antibody production, to be carried out simultaneously and continuously, thereby increasing the number of antibodies produced. Furthermore, in the circulation process, the medium is returned to the second pool P2, thereby enabling the reuse of resources required for cell culture.

[0099] In the case of continuous operation as in this embodiment, the first medium and the second medium may be supplied to the first pool P1 and the second pool P2 as needed during operation. Alternatively, only the nutrients in the first medium and the second medium may be supplied as needed. This allows the nutrients necessary for the cells to be maintained, enabling continuous operation without slowing down the culture rate.

[0100] Fourth Embodiment Next, a bioreactor system 1 according to a fourth embodiment will be described with reference to Fig. 17 . In Fig. 17 , components similar to those in the third embodiment are denoted by the same reference numerals, and detailed description thereof will be omitted. In the third embodiment, a portion of the discharge line 111 passes through the interior of the inner cylinder 30. That is, the discharge line 111 is disposed to penetrate the lower base 21 from bottom to top, thereby reaching the interior of the inner cylinder 30. The discharge line 111 then extends to the top of the inner cylinder 30, turns back downward, and then penetrates the lower base 21 from top to bottom to reach the outside of the inner cylinder 30.

[0101] The cell acquisition unit 120 and antibody acquisition unit 130 on the discharge line 111 are provided inside the inner cylinder 30. Note that only either the cell acquisition unit 120 or the antibody acquisition unit 130 may be provided inside the inner cylinder 30. That is, it is sufficient that at least one of the cell acquisition unit 120 and the antibody acquisition unit 130 is provided inside the inner cylinder 30. Furthermore, although the extraction pump 113 and the discharge valve 112 are provided outside the inner cylinder 30, they may also be configured to be provided inside the inner cylinder 30. Furthermore, a part of the circulation line 140, the circulation valve 141, may be provided inside the inner cylinder 30.

[0102] This allows for effective use of the space inside the inner cylinder 30. In other embodiments, the entire device can be made more compact by providing components that are provided outside the casing 20 inside the inner cylinder 30. Furthermore, by consolidating and arranging various components inside the inner cylinder 30, maintainability can be improved.

[0103] Fifth Embodiment Next, a bioreactor system 1 according to a fifth embodiment will be described with reference to Fig. 18. In Fig. 18, the same components as those in the other embodiments are denoted by the same reference numerals, and detailed description thereof will be omitted.

[0104] <Sealing portion> In the fifth embodiment, a sealing portion 180 is provided to circumferentially seal between the lower end edge portion 43 a of the rotating cylinder 43 and the upper surface of the lower bottom portion 21. The sealing portion 180 has a rotating seal piece 181 and a fixed seal piece 182.

[0105] The rotary seal pieces 181 are provided around the entire circumference of the inner and outer peripheral surfaces of the rotary cylinder 43. The rotary seal pieces 181 protrude from the inner and outer peripheral surfaces of the rotary cylinder 43 and then extend downward.

[0106] A pair of stationary seal pieces 182 are provided on the upper surface of the lower bottom portion 21. The stationary seal pieces 182 are provided to correspond to the pair of rotary seal pieces 181 on the upper surface of the lower bottom portion 21. The stationary seal pieces 182 protrude upward from the lower bottom portion 21 and are provided to fit over the entire circumference between the rotating cylinder 43 and the rotary seal pieces 181. The rotary seal pieces 181 and the stationary seal pieces 182 form a labyrinth mechanism radially inside and outside the clearance.

[0107] In this embodiment, the provision of the sealing portion 180 makes it possible to suppress the movement of cells and culture medium through the clearance between the rotating cylinder 43 and the lower bottom portion 21. This allows the first pool P1 and the second pool P2 to be appropriately partitioned, ensuring that cell culture can be carried out in each pool according to the growth phase.

[0108] <First Modification of Fifth Embodiment> As a first modification of the fifth embodiment, the seal portion 180 may have the configuration shown in Fig. 19. The seal portion 180 of the first modification is composed of an annular groove 183 formed in the rotating cylinder 43 and fins 184, 186 provided on the lower bottom portion 21.

[0109] The annular groove 183 is recessed upward from the lower edge 43a of the rotating cylinder 43 and extends annularly around the entire circumference. A plurality of (two in this embodiment) annular grooves 183 are formed at radial intervals. A plurality of (two in this embodiment) fins 184, 186 are provided to correspond to the annular grooves 183. The lower end of each fin 184, 186 is fixed to the upper surface of the lower base 21 and extends upward to fit into each annular groove 183.

[0110] This allows a labyrinth mechanism to be formed within the clearance. Compared to the fourth embodiment, there are no protrusions from the inner and outer surfaces of the rotating cylinder 43, which reduces the generation of turbulence in the first pool P1 and the second pool P2. Therefore, appropriate Taylor vortices can be formed in the first pool P1 and the second pool P2.

[0111] <Second Modification of Fifth Embodiment> In addition, as a second modification of the fifth embodiment, the seal portion 180 may have the configuration shown in Fig. 20. The seal portion 180 of the second modification is composed of a notch 185 formed in the rotating cylinder 43 and fins 184, 186 provided on the lower bottom portion 21.

[0112] A pair of notches 185 are provided at the lower part of the rotating cylinder 43 so as to be recessed from the outer and inner peripheral surfaces of the rotating cylinder 43 and open downward. As a result, the lower end of the rotating cylinder 43 is formed with a smaller radial thickness than the upper part.

[0113] Similar to the first modification, the fins 184, 186 are annular and protrude from the upper surface of the lower bottom portion 21, extending over the entire circumferential direction. The fins 184, 186 are arranged to be accommodated in the respective notches 185 so as to correspond to the pair of notches 185. The inner peripheral surface of the rotating cylinder 43 and the inner peripheral surface of the radially inner fin 184, 186 of the pair of fins 184, 186 are located flush with each other on the same cylindrical surface. The outer peripheral surface of the rotating cylinder 43 and the outer peripheral surface of the radially outer fin 184, 186 of the pair of fins 184, 186 are located flush with each other on the same cylindrical surface.

[0114] This allows a labyrinth mechanism to be formed, as in the first modified example, and since no protrusions are formed into the first pool P1 and the second pool P2, appropriate Taylor vortices can be formed in the first pool P1 and the second pool P2.

[0115] Sixth Embodiment Next, a bioreactor system 1 according to a sixth embodiment will be described with reference to Fig. 21. In Fig. 21, components similar to those in the other embodiments are denoted by the same reference numerals, and detailed description thereof will be omitted.

[0116] <Low Thermal Conductivity Member> In the sixth embodiment, a low thermal conductivity member 190 is provided on the surface of the rotating cylinder 43. The low thermal conductivity member 190 is provided along the inner and outer circumferential surfaces of the rotating cylinder 43. The low thermal conductivity member 190 is provided over at least the entire area of ​​the inner and outer circumferential surfaces of the rotating cylinder 43 that come into contact with the culture solution. The low thermal conductivity member 190 may be provided on at least one of the inner and outer circumferential surfaces of the rotating cylinder 43.

[0117] The low thermal conductivity member 190 is made of a material having a lower thermal conductivity than the material that constitutes the rotating cylinder 43. For example, if the low thermal conductivity member 190 is made of a steel material such as stainless steel, the low thermal conductivity member 190 may be made of ceramic applied by thermal spraying, separately attached rubber, or the like.

[0118] The presence of the low thermal conductivity member 190 can suppress heat transfer between the first pool P1 and the second pool P2. This allows the first temperature adjustment unit 91 and the second top heating unit to more appropriately control the temperatures of the first pool P1 and the second pool P2 independently. This allows more appropriate cell culture according to the growth phase in the first pool P1 and the second pool P2.

[0119] In order to suppress heat transfer between the first pool P1 and the second pool P2, for example, the material constituting the rotating cylinder 43 may itself be made of a material having a lower thermal conductivity than the outer cylinder 22. This allows the second temperature adjustment unit 92 to appropriately adjust the temperature inside the second pool P2 from the outside of the outer cylinder 22, while suppressing heat transfer between the first pool P1 and the second pool P2, and allowing the temperatures in each pool to be set independently.

[0120] Other Embodiments Although the embodiments of the present invention have been described above, the present invention is not limited to these and can be modified as appropriate within the scope of the technical concept of the invention.

[0121] For example, although the operation method of the bioreactor 10 in the embodiment has been described as being configured to be performed by the control device 200, all or some of the steps may be performed by an operator. For example, the medium supply step, cell supply step, transfer step, circulation step, and discharge step may be performed manually by an operator. Furthermore, the temperature setting unit 202 may receive input from the operator and control the first temperature adjustment unit and the second temperature adjustment unit in accordance with the input. Furthermore, the rotation control unit 213 may receive input from the operator and rotate the rotating cylinder 43 via the drive unit 50 in accordance with the input. Similarly, the oxygen supply unit 100 may receive input from the operator and control the oxygen supply unit 100 in accordance with the input.

[0122] In the embodiment, cells grown in the first pool P1 were moved to the second pool P2 and used, but simultaneous culture may also be performed by supplying cells in different growth phases to the first pool P1 and the second pool P2 from the outside.

[0123] <Hardware Configuration> The processing steps performed by the control device 200 described above are stored in the form of a program on a computer-readable recording medium, and the computer reads and executes this program to perform the above processing. Specific examples of computers are shown below.

[0124] As shown in Fig. 22, the computer includes a CPU 301, a main memory 302, a storage 303, and an interface 304. For example, the above-mentioned arithmetic device is implemented in the computer. The operations of the above-mentioned processing units are stored in the storage 303 in the form of a program. The CPU 301 reads the program from the storage 303, loads it into the main memory 302, and executes the above-mentioned processing in accordance with the program. The CPU 301 also allocates a storage area in the main memory 302 in accordance with the program.

[0125] Examples of storage 303 include a hard disk drive (HDD), a solid state drive (SSD), a magnetic disk, a magneto-optical disk, a compact disc read-only memory (CD-ROM), a digital versatile disc read-only memory (DVD-ROM), and a semiconductor memory. Storage 303 may be an internal medium directly connected to the computer's bus, or an external medium connected to the computer via interface 304 or a communication line. Furthermore, when this program is distributed to a computer via a communication line, the computer that receives the program may load the program into main memory 302 and execute the above-described processing. Note that storage 303 is a non-transitory tangible storage medium.

[0126] The program may also implement some of the functions described above. Furthermore, the program may be a file that can implement the functions described above in combination with a program already stored on the computer, a so-called differential file (differential program).

[0127] In addition to or instead of the above configuration, a custom LSI (Large Scale Integrated Circuit) such as a PLD (Programmable Logic Device), an ASIC (Application Specific Integrated Circuit), a GPU (Graphics Processing Unit), or a processing device similar thereto may be provided. Examples of PLDs include programmable array logic (PAL), generic array logic (GAL), complex programmable logic device (CPLD), and field programmable gate array (FPGA). In this case, some or all of the functions realized by a processor may be realized by the integrated circuit.

[0128] <Notes> The descriptions in each embodiment can be understood as follows, for example.

[0129] (1) A bioreactor 10 according to a first aspect includes an outer cylinder 22 centered on an axis O extending in the vertical direction, an inner cylinder 30 provided inside the outer cylinder 22 and centered on the axis O, a rotating cylinder 43 disposed between the outer cylinder 22 and the inner cylinder 30, defining a first pool P1 between the outer cylinder 22 and the inner cylinder 30 and defining a second pool P2 between the outer cylinder 22 and the inner cylinder 30, and being rotatable about the axis O, a drive unit 50 that drives the rotating cylinder 43 to rotate about the axis O, a first supply port 61 a capable of supplying a liquid to the first pool P1, and a second supply port 71 a capable of supplying a liquid to the second pool P2.

[0130] This allows different media to be supplied to the first pool P1 and the second pool P2. Furthermore, by rotating the rotating cylinder 43, Taylor vortices can be formed in both the first pool P1 and the second pool P2. This allows cells in different growth phases to be cultured simultaneously in a compact configuration.

[0131] (2) The bioreactor 10 according to the second aspect is the bioreactor 10 of (1), which includes a first culture medium supply unit 60 capable of supplying the first culture medium as the liquid to the first pool P1 via the first supply port 61 a, and a second culture medium supply unit 70 capable of supplying a second culture medium having components different from the first culture medium as the liquid to the second pool P2 via the second supply port 71 a.

[0132] This allows for the creation of growth environments for cells in different growth phases in each of the first pool P1 and the second pool P2.

[0133] (3) The bioreactor 10 according to the third aspect is the bioreactor 10 of (1) or (2), which is equipped with a first temperature control unit 91 capable of adjusting the temperature of the first pool P1 and a second temperature control unit 92 capable of adjusting the temperature of the second pool P2 independently of the first temperature control unit 91.

[0134] This allows the first pool P1 and the second pool P2 to be set at appropriate temperatures according to the growth phase of the cells.

[0135] (4) The bioreactor 10 according to the fourth aspect is the bioreactor 10 of (3), which is provided with a low thermal conductivity member 190 provided on the surface of the rotating cylinder 43 and having a thermal conductivity lower than that of the rotating cylinder 43.

[0136] This makes it possible to suppress heat transfer between the first pool P1 and the second pool P2.

[0137] (5) A bioreactor 10 according to a fifth aspect is the bioreactor 10 according to (3) or (4), in which the rotating cylinder 43 is made of a material having a lower thermal conductivity than the outer cylinder 22.

[0138] This makes it possible to suppress heat transfer between the first pool P1 and the second pool P2.

[0139] (6) The bioreactor 10 according to a sixth aspect is any one of the bioreactors 10 of (1) to (5) including an oxygen supply unit 100 capable of supplying oxygen to the first pool P1 and the second pool P2.

[0140] This allows the first pool P1 and the second pool P2 to be supplied with oxygen necessary for cell culture.

[0141] (7) A seventh aspect of the bioreactor 10 is the bioreactor 10 of (6), in which the oxygen supply unit 100 includes a first oxygen supply unit 101 capable of supplying oxygen to the first pool P1 and a second oxygen supply unit 102 capable of supplying oxygen to the second pool P2 independently of the first oxygen supply unit 101.

[0142] This allows appropriate oxygen supply amounts to be set according to the growth phases of the cells in the first pool P1 and the second pool P2.

[0143] (8) The bioreactor 10 according to the eighth aspect is a bioreactor 10 according to any one of (1) to (7), which is provided with a cell supply unit 80 that supplies cells to the first pool P1, and a moving mechanism 150 that moves at least a portion of the liquid in the first pool P1 to the second pool P2.

[0144] This allows, for example, after cells have been grown in the first pool P1, when the cells progress to the next growth phase, to be moved to the second pool P2, allowing for culturing appropriate for that growth phase.

[0145] (9) A ninth aspect of the bioreactor 10 is the bioreactor 10 of (8), in which the moving mechanism 150 has a moving pipe connecting the first pool P1 and the second pool P2, and a moving valve 152 provided in the moving pipe and capable of being opened and closed.

[0146] This allows the medium and cells to be transferred from the first pool P1 to the second pool P2.

[0147] (10) The bioreactor 10 according to the tenth aspect is the bioreactor 10 of (8), in which the drive unit 50 is capable of driving the rotating cylinder 43 to rotate forward and backward, the rotating cylinder 43 has an opening 43b that penetrates the rotating cylinder 43 radially, and the moving mechanism 150 has an opening / closing unit that closes the opening 43b when the rotating cylinder 43 rotates forward and opens the opening 43b when the rotating cylinder 43 rotates backward.

[0148] This allows the medium and cells to be easily transferred from the first pool P1 to the second pool P2.

[0149] (11) The bioreactor 10 according to the eleventh aspect is the bioreactor 10 of (10), in which the moving mechanism 150 has a sliding door 160 that can slide between a closed position that closes the opening 43b and an open position that is located forward of the closed position in the forward rotation direction and opens the opening 43b.

[0150] This allows the medium and cells to be easily transferred from the first pool P1 to the second pool P2.

[0151] (12) The bioreactor 10 according to the twelfth aspect is the bioreactor 10 of (10), in which the moving mechanism 150 has an opening / closing door 170 that is rotatable around a rotation axis extending vertically at the front side of the opening 43b in the forward rotation direction, and the opening / closing door 170 is rotatable between a closed position that closes the opening 43b and an open position that moves away from the opening 43b to open the opening 43b.

[0152] This allows the medium and cells to be easily transferred from the first pool P1 to the second pool P2.

[0153] (13) The bioreactor 10 according to the thirteenth aspect is any of the bioreactors 10 of (1) to (12), further comprising an extraction unit 110 having a discharge line 111 capable of extracting the liquid in the second pool P2 to the outside, a cell acquisition unit 120 provided in the discharge line 111 for extracting the cells from the liquid, and an antibody acquisition unit 130 provided in the discharge line 111 for extracting antibodies produced by the cells from the liquid.

[0154] This allows the liquid to be removed from the second pool P2, and the cells and antibodies to be extracted from the liquid.

[0155] (14) The bioreactor 10 according to the fourteenth aspect is the bioreactor 10 of (13), in which the discharge line 111 passes through the inside of the inner cylinder 30, and at least one of the cell acquisition section 120 and the antibody acquisition section 130 is provided inside the inner cylinder 30.

[0156] This allows the space inside the inner cylinder 30 to be used effectively, and the installation space for the cell acquisition section 120 and the antibody acquisition section 130 to be used more efficiently.

[0157] (15) The bioreactor 10 according to the fifteenth aspect is the bioreactor 10 of (13) or (14), in which the removal section 110 is connected to the discharge line 111 downstream of the cell acquisition section 120 and the antibody acquisition section 130 and has a circulation line 140 capable of returning the liquid to the second pool P2.

[0158] This allows for the reuse of the medium and cells, and also allows for continuous culture, which improves the efficiency of antibody production.

[0159] (16) The bioreactor 10 according to the sixteenth aspect is any one of the bioreactors 10 of (1) to (15), which has a lower bottom 21 that closes the outer cylinder 22 from below, and the lower edge 43a of the rotating cylinder 43 faces the lower end of the lower bottom 21 via a clearance.

[0160] This allows the rotating cylinder 43 to rotate smoothly while preventing the liquids in the first pool P1 and the second pool P2 from accidentally mixing with each other.

[0161] (17) The bioreactor 10 according to the seventeenth aspect is the bioreactor 10 of (16), further comprising a seal portion 180 that seals between the lower edge portion 43 a of the rotating cylinder 43 and the upper surface of the lower bottom portion 21.

[0162] This makes it possible to further prevent the liquids in the first pool P1 and the second pool P2 from mixing with each other.

[0163] (18) A bioreactor system 1 according to an eighteenth aspect includes the bioreactor 10 of (1) and a control device 200 that controls the bioreactor 10. The bioreactor 10 includes a first culture medium supply unit 60 that can supply a first culture medium as the liquid to the first pool P1 via the first supply port 61 a, and a second culture medium supply unit 70 that can supply a second culture medium having components different from the first culture medium as the liquid to the second pool P2 via the second supply port 71 a. The control device 200 includes a culture medium supply control unit 201 that executes a culture medium supply step of controlling the first culture medium supply unit 60 to supply the first culture medium to the first pool P1 and controlling the second culture medium supply unit 70 to supply the second culture medium to the second pool P2.

[0164] This makes it possible to create a growth environment for cells in different growth phases in a compact configuration.

[0165] (19) A bioreactor system 1 according to a 19th aspect is the bioreactor system 1 of (18), in which the bioreactor 10 has a cell supply unit 80 that supplies cells to the culture medium of the first pool P1, and the control device 200 has a cell supply control unit 203 that controls the cell supply unit 80 after the culture medium supply step to execute a cell supply step of supplying the cells to the first pool P1, and a culture execution unit 204 that controls the drive unit 50 after the cell supply step to execute a culture step of rotating the rotating cylinder 43.

[0166] This allows cell proliferation in the first pool P1.

[0167] (20) A bioreactor system 1 according to a twentieth aspect is a bioreactor system 1 of (19), in which the bioreactor 10 has a moving mechanism 150 that moves at least a portion of the liquid in the first pool P1 to the second pool P2, the control device 200 has a moving execution unit 205 that controls the moving mechanism 150 to execute a moving process of moving the liquid in the first pool P1 to the second pool P2 after the culture process, and the culture execution unit 204 executes the culture process again after the moving process.

[0168] By transferring cells grown in the first pool P1 to the second pool P2, it is possible to sequentially culture the cells according to their growth phases in a single device. For example, in the second phase, an environment suitable for antibody production by the cells is created, allowing for efficient antibody production.

[0169] (21) A bioreactor system 1 according to a 21st aspect is a bioreactor system 1 (20) in which the moving mechanism 150 has a moving pipe connecting the first pool P1 and the second pool P2, and a moving valve 152 provided in the moving pipe and capable of being opened and closed, and the moving execution unit 205 moves the liquid in the first pool P1 to the second pool P2 by controlling the moving valve 152 as the moving process.

[0170] This allows easy transfer of cells from the first pool P1 to the second pool P2.

[0171] (22) A bioreactor system 1 according to a 22nd aspect is the bioreactor system 1 of (20), wherein the drive unit 50 is capable of driving the rotating cylinder 43 in forward and reverse rotation, the culture execution unit 204 controls the drive unit 50 to drive the rotating cylinder 43 in forward rotation, the rotating cylinder 43 has an opening 43b that penetrates the rotating cylinder 43 in the radial direction, the movement mechanism 150 has an opening / closing unit that closes the opening 43b when the rotating cylinder 43 rotates forward and opens the opening 43b when the rotating cylinder 43 rotates reversely, and the movement execution unit 205 controls the drive unit 50 to drive the rotating cylinder 43 in reverse rotation at a rotation speed lower than the forward rotation speed as the movement process.

[0172] As a result, the cells and culture medium in the first pool P1 can be moved to the second pool P2 simply by rotating the rotating cylinder 43. That is, since the cells can be moved by controlling only the rotating cylinder 43, a simple configuration can be achieved.

[0173] (23) A bioreactor system 1 according to a 23rd aspect is a bioreactor system 1 of any of (20) to (22), in which the bioreactor 10 includes a discharge line 111 capable of discharging the liquid in the second pool P2 to the outside, a cell acquisition unit 120 provided in the discharge line 111 to extract the cells from the liquid, and an extraction unit 110 having an antibody acquisition unit 130 provided in the discharge line 111 to extract antibodies produced by the cells from the liquid, and the control device 200 includes a discharge execution unit 206 that controls the extraction unit 110 to execute a discharge step of extracting and discharging the liquid from the second pool P2 after the culture step following the transfer step is completed.

[0174] This allows the liquid in the second pool P2 after antibody production to be removed to the outside, and allows cells and antibodies to be extracted from the liquid.

[0175] (24) In a bioreactor system 1 according to a 24th aspect, the take-out unit 110 has a circulation line 140 that is connected to the discharge line 111 downstream of the cell acquisition unit 120 and the antibody acquisition unit 130 and that can return the liquid to the second pool P2, the cell supply control unit 203 executes the cell supply step again between the transfer step and the culture step after the transfer step, and the control device 200 determines whether to perform continuous operation between the culture step after the transfer step and the discharge step. The discharge execution unit 206 executes the discharge step when the determination unit 208 determines that continuous operation is not to be performed, and the discharge execution unit 206 executes the discharge step when the determination unit 208 determines that continuous operation is to be performed, and the discharge execution unit 206 executes the circulation step of returning the liquid extracted from the second pool P2 to the second pool P2 via the circulation line 140 by controlling the extraction unit 110 when the determination unit 208 determines that continuous operation is to be performed, and the transfer execution unit 205 executes the transfer step again after the circulation step (23).

[0176] This allows the medium to be reused. Furthermore, by repeating the circulation process, cell growth in the first pool P1 and antibody production in the second pool P2 can be carried out sequentially. In other words, continuous culture is possible, which improves the efficiency of antibody production.

[0177] (25) A bioreactor system 1 according to a 25th aspect includes the bioreactor 10 of (1) and a control device 200 for controlling the bioreactor 10, wherein the bioreactor 10 includes a first temperature control unit 91 capable of adjusting the temperature of the first pool P1 and a second temperature control unit 92 capable of adjusting the temperature of the second pool P2, and the control device 200 includes a temperature setting unit 202 for controlling the first temperature control unit 91 and the second temperature control unit 92 so that the first pool P1 and the second pool P2 have different temperatures.

[0178] This allows the first pool P1 and the second pool P2 to be set to appropriate temperatures separately.

[0179] (26) A bioreactor system 1 according to a 26th aspect includes the bioreactor 10 of (1) and a control device 200 that controls the bioreactor 10. The bioreactor 10 includes a first oxygen supply unit 101 that can supply oxygen to the first pool P1 and a second oxygen supply unit 102 that can supply oxygen to the second pool P2. The control device 200 has an oxygen supply control unit 212 that controls the first oxygen supply unit 101 and the second oxygen supply unit 102 so that the amounts of oxygen supplied to the first pool P1 and the second pool P2 are different from each other.

[0180] This allows the oxygen concentrations in the first pool P1 and the second pool P2 to be set to appropriate values ​​separately.

[0181] (27) A twenty-seventh aspect of the present invention relates to a method for operating a bioreactor (10) comprising: an outer cylinder (22) centered on an axis O extending in the vertical direction; an inner cylinder (30) centered on the axis O inside the outer cylinder (22); a rotating cylinder (43) disposed between the outer cylinder (22) and the inner cylinder (30) to define a first pool (P1) between the outer cylinder (22) and the inner cylinder (30) and to define a second pool (P2) between the outer cylinder (22) and the inner cylinder (30), the rotating cylinder (43) being rotatable about the axis O; and a drive unit (50) that drives the rotating cylinder (43) to rotate about the axis O, the method comprising a culture medium supply step of supplying a first culture medium to the first pool (P1) and supplying a second culture medium having components different from those of the first culture medium to the second pool (P2).

[0182] This makes it possible to culture cells according to their growth phase with a compact configuration.

[0183] (28) A 28th aspect of the method for operating the bioreactor 10 is the method for operating the bioreactor 10 of (27), further including a cell supplying step of supplying the cells to the first pool P1 after the culture medium supplying step, and a culturing step of rotating the rotating cylinder 43 via the driving unit 50 after the cell supplying step.

[0184] This allows cells to grow in the first pool P1.

[0185] (29) A 29th aspect of the method for operating a bioreactor 10 is the method for operating a bioreactor 10 of (28), further including a transfer step of transferring the liquid in the first pool P1 to the second pool P2 after the culture step, and the culture step is executed again after the transfer step.

[0186] By transferring the cells grown in the first pool P1 to the second pool P2, culture can be performed according to the cell growth phase. For example, in the second phase, an environment suitable for antibody production by the cells can be created, allowing for efficient antibody production.

[0187] (30) A thirtieth aspect of the method for operating a bioreactor 10 is the method for operating a bioreactor 10 of (29), further including a discharge step of removing the liquid from the second pool P2 and extracting the cells and the antibodies produced by the cells from the liquid after the culture step following the transfer step is completed.

[0188] This allows the liquid in the second pool P2 after antibody production to be removed to the outside, and allows cells and antibodies to be extracted from the liquid.

[0189] (31) A thirty-first aspect of the method for operating a bioreactor (10) according to the thirty-first aspect includes a determination step for determining whether or not to perform continuous operation between the culturing step and the discharging step after the transferring step, the discharging step being executed when it is determined in the determination step that continuous operation is not to be performed, and further includes a circulation step for removing liquid from the second pool (P2), extracting the cells and the antibodies from the liquid, and then returning the liquid to the second pool (P2), when it is determined in the determination step that continuous operation is to be performed, and the method for operating a bioreactor (10) according to (30), wherein the transferring step is executed again after the circulation step.

[0190] This allows the medium to be reused. Furthermore, by repeating the circulation process, cell growth in the first pool P1 and antibody production in the second pool P2 can be carried out sequentially. In other words, continuous culture is possible, which improves the efficiency of antibody production.

[0191] According to the present disclosure, it is possible to carry out cell culture according to the growth phase with a compact configuration.

[0192] REFERENCE SIGNS LIST 1 Bioreactor system 10 Bioreactor 20 Casing 21 Lower bottom 22 Outer cylinder 23 Lid 23a Penetration section 30 Inner cylinder 40 Rotating section 41 Shaft 42 Upper bottom 43 Rotating cylinder 43a Lower edge 43b Opening 50 Drive section 60 First culture medium supply section 61 First supply pipe 61a First supply port 62 First supply line 63 First supply valve 64 First culture medium storage section 70 Second culture medium supply section 71 Second supply pipe 71a Second supply port 72 Second supply line 73 Second supply valve 74 Second culture medium storage section 80 Cell supply section 81 Cell supply pipe 81a Cell supply port 82 Cell supply line 83 Cell supply valve 84 Cell storage section 91 First temperature adjustment section 92 Second temperature adjustment unit 100 Oxygen supply unit 101 First oxygen supply unit 102 Second oxygen supply unit 110 Take-out unit 111 Discharge line 112 Discharge valve 113 Take-out pump 120 Cell acquisition unit 130 Antibody acquisition unit 140 Circulation line 141 Circulation valve 150 Movement mechanism 151 Movement line 152 Movement valve 160 Sliding door 161 Stopper 170 Opening / closing door 171 Hinge 180 Seal unit 181 Rotating seal piece 182 Fixed seal piece 183 Annular groove 184 Fin 185 Notch 186 Fin 190 Low thermal conductivity member 200 Control device 201 Culture medium supply control unit 202 Temperature setting unit 203 Cell supply control unit 204 Cultivation execution unit 205 Movement execution unit 206 Discharge execution unit 207 Circulation execution unit 208 Determination unit 211 Valve control unit 212 Oxygen supply control unit 213 Rotation control unit 214 Pump control unit 301 CPU 302 Main memory 303 Storage 304 Interface O Axis P1 First pool P2 Second pool

Claims

1. A bioreactor comprising: an outer cylinder centered on an axis extending in a vertical direction; an inner cylinder disposed inside the outer cylinder and centered on the axis; a rotating cylinder disposed between the outer cylinder and the inner cylinder, defining a first pool between the outer cylinder and the inner cylinder and defining a second pool between the outer cylinder and the inner cylinder, and capable of rotating about the axis; a drive unit that drives the rotating cylinder to rotate about the axis; a first supply port capable of supplying liquid to the first pool; and a second supply port capable of supplying liquid to the second pool.

2. The bioreactor according to claim 1, comprising: a first culture medium supply unit capable of supplying a first culture medium as the liquid to the first pool via the first supply port; and a second culture medium supply unit capable of supplying a second culture medium having components different from the first culture medium as the liquid to the second pool via the second supply port.

3. A bioreactor as described in claim 1, comprising: a first temperature control unit capable of adjusting the temperature of the first pool; and a second temperature control unit capable of adjusting the temperature of the second pool independently of the first temperature control unit.

4. The bioreactor according to claim 1, further comprising an oxygen supply unit capable of supplying oxygen to the first pool and the second pool.

5. The bioreactor described in claim 4, wherein the oxygen supply unit includes: a first oxygen supply unit capable of supplying oxygen to the first pool; and a second oxygen supply unit capable of supplying oxygen to the second pool independently of the first oxygen supply unit.

6. A bioreactor described in any one of claims 1 to 5, comprising: a cell supply unit that supplies cells to the first pool; and a movement mechanism that moves at least a portion of the liquid in the first pool to the second pool.

7. A bioreactor as described in claim 6, wherein the transfer mechanism comprises: a transfer pipe connecting the first pool and the second pool; and a transfer valve provided in the transfer pipe and capable of being opened and closed.

8. A bioreactor as described in claim 6, wherein the drive unit is capable of driving the rotating cylinder to rotate in both forward and reverse directions, the rotating cylinder has an opening that penetrates the rotating cylinder in the radial direction, and the moving mechanism has an opening / closing unit that closes the opening when the rotating cylinder rotates in the forward direction and opens the opening when the rotating cylinder rotates in the reverse direction.

9. A bioreactor as described in claim 8, wherein the moving mechanism has a sliding door that can slide between a closing position that closes the opening and an opening position that is located forward of the closing position in the forward rotation direction and opens the opening.

10. A bioreactor as described in claim 8, wherein the moving mechanism has an opening / closing door that is rotatable around a rotation axis extending vertically at the front side of the opening in the positive rotation direction, and the opening / closing door is rotatable between a closing position that closes the opening and an opening position that moves away from the opening to open the opening.

11. A bioreactor described in any one of claims 1 to 5, further comprising an extraction section having a discharge line capable of removing the liquid in the second pool to the outside, a cell acquisition section provided in the discharge line for extracting cells from the liquid, and an antibody acquisition section provided in the discharge line for extracting antibodies produced by the cells from the liquid.

12. A bioreactor as described in claim 11, wherein the discharge line passes through the inside of the inner cylinder, and at least one of the cell acquisition section and the antibody acquisition section is provided inside the inner cylinder.

13. A bioreactor as described in claim 11, wherein the removal section has a circulation line connected to the discharge line downstream of the cell acquisition section and the antibody acquisition section and capable of returning the liquid to the second pool.

14. A bioreactor described in any one of claims 1 to 5, further comprising a lower bottom portion that closes the outer cylinder from below, and wherein the lower edge portion of the rotating cylinder faces an upper surface of the lower bottom portion via a clearance.

15. The bioreactor of claim 14, further comprising a seal portion for sealing between the lower edge of the rotating cylinder and the upper surface of the lower bottom portion.

16. A bioreactor system comprising: a bioreactor as described in claim 1; and a control device for controlling the bioreactor, wherein the bioreactor comprises: a first culture medium supply unit capable of supplying a first culture medium as the liquid to the first pool via the first supply port; and a second culture medium supply unit capable of supplying a second culture medium having components different from the first culture medium as the liquid to the second pool via the second supply port, and the control device has a culture medium supply control unit that executes a culture medium supply step of controlling the first culture medium supply unit to supply the first culture medium to the first pool and controlling the second culture medium supply unit to supply the second culture medium to the second pool.

17. A method for operating a bioreactor comprising: an outer cylinder centered on an axis extending in the vertical direction; an inner cylinder centered on the axis inside the outer cylinder; a rotating cylinder disposed between the outer cylinder and the inner cylinder, defining a first pool between the inner cylinder and defining a second pool between the outer cylinder and the inner cylinder, and capable of rotating about the axis; and a drive unit that drives the rotating cylinder to rotate about the axis, the method including a culture medium supplying step of supplying a first culture medium to the first pool and supplying a second culture medium having components different from the first culture medium to the second pool.

18. The method for operating a bioreactor described in claim 17, further comprising: a cell supplying step of supplying cells to the first pool after the culture medium supplying step; and a culturing step of rotating the rotating cylinder via the driving unit after the cell supplying step.

19. The method for operating a bioreactor according to claim 18, further comprising a transfer step of transferring the liquid in the first pool to the second pool after the culturing step, wherein the culturing step is carried out again after the transfer step.

20. A method for operating a bioreactor as described in claim 19, further comprising a discharge step of removing the liquid from the second pool and extracting the cells and antibodies produced by the cells from the liquid after the culture step following the transfer step is completed.

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