Cultivation apparatus

Abstract

To provide a cultivation apparatus which is excellent in the efficiency of adjusting the concentration of an adjustment target dissolved gas in a culture solution and which is less likely to cause damage to a cultivation target.SOLUTION: A cultivation apparatus comprises: a circulation path having a reactor for cultivating a cultivation target; a pump for circulating a culture solution containing the cultivation target in the circulation path; a gas concentration adjustment unit forming a branch path branched from a branch portion of the circulation path and joined to a junction portion of the circulation path; a supply path for supplying a supply gas from a gas supply source to the branch path; and a discharge path for discharging the supply gas from the branch path. The gas concentration adjustment unit includes a gas-permeable member and a filter. The gas-permeable member adjusts the concentration of the adjustment target dissolved gas in the culture solution in the branch path by bringing the supply gas supplied from the supply path into contact with the culture solution through the gas-permeable member.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to an incubation device. [Background technology]

[0002] A culture device is known that has a circulation path with a reactor for cultivating the culture target, a pump for circulating a culture solution containing the culture target within the circulation path, and a gas concentration adjustment unit that adjusts the concentration of a dissolved gas to be adjusted in the culture solution (see, for example, Patent Documents 1 and 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent No. 7216239 [Patent Document 2] Japanese Patent Application Laid-Open No. 2012-100547 Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a culture apparatus which is excellent in the efficiency of adjusting the concentration of a dissolved gas to be adjusted in a culture solution and which is less likely to damage the culture object. [Means for solving the problem]

[0005] One aspect of the present invention is as follows.

[0006] [1] a circulation path having a reactor for cultivating a culture target; a pump that circulates the culture solution containing the culture subject through the circulation path; a gas concentration adjusting unit that forms a branch path that branches from the branching portion of the circulation path and joins the joining portion of the circulation path; a supply path for supplying a supply gas from a gas supply source to the branch path; an exhaust path for exhausting the supply gas from the branch path, the gas concentration adjusting unit includes a gas permeable member and a filter that is provided between the branching unit and the gas permeable member and that prevents the culture object from passing through and allows the culture solution to pass through; The gas-permeable member adjusts the concentration of the dissolved gas to be adjusted in the culture solution in the branch path by bringing the supply gas supplied from the supply path into contact with the culture solution via the gas-permeable member.

[0007] [2] The culture apparatus according to [1], further comprising a flow rate adjusting unit that adjusts the flow rate of the culture solution toward the circulation path side and the flow rate of the culture solution toward the branch path side at the branching section.

[0008] [3] The culture device according to [2], wherein the flow rate adjusting unit allows the culture medium to flow simultaneously on both the circulation path side and the branch path side.

[0009] [4] a sensor for measuring the concentration of the dissolved gas to be adjusted in the culture solution; The culture device according to [2] or [3], further comprising a control device electrically connected to the sensor and the three-way valve as the flow rate control unit, and controlling the three-way valve in accordance with the measurement results of the sensor.

[0010] [5] The culture device according to any one of [1] to [4], wherein the gas concentration adjusting unit has a check valve that allows the culture solution to flow from the branching part to the confluence part in the branching path while suppressing the flow in the reverse direction.

[0011] [6] The culture apparatus according to any one of [1] to [5], wherein the branching section and the confluence section are located inside the reactor.

[0012] [7] The culture apparatus according to any one of [1] to [6], wherein the supply gas supplied through the supply path contains carbon dioxide, and the dissolved gas to be adjusted contains carbon dioxide.

[0013] [8] The culture apparatus according to any one of [1] to [7], wherein the supply gas supplied through the supply path contains nitrogen, and the dissolved gas to be adjusted contains oxygen.

[0014] [9] a supply step of adjusting the concentration of the dissolved gas to be adjusted in the culture solution in the branch path by bringing the pressurized supply gas supplied from the supply path into contact with the culture solution via the gas permeable member; a discharge step of discharging the pressurized supply gas from the branch passage to reduce the pressure of the supply gas in the branch passage; A method for operating a culture apparatus according to any one of [1] to [8], wherein a confluence process is carried out in this order, in which the culture solution in which the concentration of the dissolved gas to be adjusted has been adjusted is returned from the branch path to the confluence part of the circulation path.

[0015]

[10] the supply path has a first on-off valve, the discharge path has a second on-off valve; the gas concentration adjusting unit includes a third on-off valve provided between the branching unit and the gas permeable member, and a fourth on-off valve provided between the gas permeable member and the junction unit, the supplying step is performed by opening the first on-off valve and closing the second on-off valve, the third on-off valve, and the fourth on-off valve, the discharge step is performed by opening the second on-off valve and closing the first on-off valve, the third on-off valve, and the fourth on-off valve, The method for operating a culture apparatus according to [9], wherein the confluence step is carried out by opening the third on-off valve and the fourth on-off valve and closing the first on-off valve. [Effects of the Invention]

[0016] According to the present invention, it is possible to provide a culture device that is excellent in the efficiency of adjusting the concentration of a dissolved gas to be adjusted in a culture solution and that is less likely to damage the culture target. [Brief explanation of the drawings]

[0017] [Figure 1] FIG. 10 is a schematic diagram showing the state of the culture device during the supply step in one embodiment of the present invention. [Figure 2] FIG. 2 is a schematic diagram showing the state of the culture device in FIG. 1 during a discharge step. [Figure 3] FIG. 2 is a schematic diagram showing the state of the culture apparatus of FIG. 1 during a confluence step. [Figure 4] 1. FIG. 4 is a schematic diagram showing another example of the gas concentration adjusting unit in FIG. [Figure 5] 1. FIG. 4 is a schematic diagram showing still another example of the gas concentration adjusting unit in FIG. [Figure 6] 2 is a schematic diagram showing an example of a gas concentration adjusting tank of the gas concentration adjusting unit of FIG. 1. FIG. [Figure 7] 2 is a schematic view showing an example of a gas permeable member of the gas concentration adjusting section of FIG. 1. FIG. [Figure 8] FIG. 2 is an explanatory diagram illustrating a method of operating the culture apparatus of FIG. DETAILED DESCRIPTION OF THE INVENTION

[0018] Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

[0019] As shown in Figures 1 to 3, in one embodiment of the present invention, a culture apparatus 1 includes a circulation path 2 having a reactor 2a for culturing the culture target, a pump 4 for circulating a culture solution 3 containing the culture target within the circulation path 2, a gas concentration adjusting unit 6 forming a branch path 5 that branches off from a branch point 2b of the circulation path 2 and joins at a confluence point 2c of the circulation path 2, a supply path 8 for supplying a supply gas from a gas supply source 7 to the branch path 5, and an exhaust path 9 for exhausting the supply gas from the branch path 5. The gas concentration adjusting unit 6 includes a gas permeable member 6a and a (first) filter 6b that is provided between the branch point 2b and the gas permeable member 6a and that prevents the culture target from passing through but allows the culture solution 3 to pass through. The gas permeable member 6a adjusts the concentration of the dissolved gas to be adjusted in the culture solution 3 in the branch path 5 by bringing the supply gas supplied from the supply path 8 into contact with the culture solution 3 via the gas permeable member 6a.

[0020] According to the above configuration, the supply gas is supplied from the supply path 8 to the branch path 5 at an appropriate pressure, and the supply gas and the culture solution 3 are brought into contact with each other through the gas-permeable member 6a, thereby adjusting the concentration of the dissolved gas to be adjusted in the culture solution 3. This allows the concentration of the dissolved gas to be adjusted to be adjusted efficiently while suppressing the consumption of the supply gas. Furthermore, the culture solution 3 passes through the filter 6b, which prevents the culture solution 3 from passing through, and reaches the gas-permeable member 6a in the branch path 5 (i.e., carbon dioxide is supplied via the gas-permeable member 6a to the culture solution 3, which is substantially isolated from the culture solution by the branch path 5 and the filter 6b). Therefore, even if the supply gas is supplied at excessive pressure and bubbles form in the culture solution 3, damage to the culture solution caused by the bubbles can be suppressed. Therefore, a culture device 1 can be realized that is highly efficient in adjusting the concentration of the dissolved gas to be adjusted in the culture solution 3 and is less likely to damage the culture solution, thereby improving, for example, the yield (and quality) of the culture solution. In order to enhance the effect of suppressing damage to the culture object, the gas concentration adjusting section 6 may be configured to have a second filter 6c, as in this embodiment, which is provided between the confluence section 2c and the gas permeable member 6a and prevents the culture object from passing through but allows the culture solution 3 to pass through.

[0021] The culture device 1 has a flow rate adjuster 10 that adjusts the flow rate of the culture solution 3 toward the circulation path 2 side and the flow rate of the culture solution 3 toward the branch path 5 side at the branch portion 2b. According to the above configuration, the flow rate adjuster 10 can cause an appropriate amount of culture solution 3 corresponding to the required adjustment amount of the concentration of the dissolved gas to be adjusted in the culture solution 3 to flow from the circulation path 2 to the gas concentration adjuster 6. It is preferable that the flow rate adjuster 10 be configured so that the culture solution 3 can flow toward both the circulation path 2 side and the branch path 5 side at the same time, as in this embodiment.

[0022] The culture apparatus 1 includes a (first) sensor 11 that measures the concentration of a dissolved gas to be adjusted in the culture solution 3, and a control device 12 that is electrically connected to the sensor 11 and a three-way valve serving as a flow rate regulator 10 and controls the three-way valve in accordance with the measurement results from the sensor 11. According to the above configuration, the control device 12, which is comprised of a computer or the like, controls the three-way valve in accordance with the concentration of the dissolved gas to be adjusted in the culture solution 3, thereby allowing an appropriate amount of the culture solution 3 to flow from the circulation path 2 to the gas concentration regulator 6. The sensor 11 may be configured to be provided downstream of the three-way valve in the reactor 2a, as in this embodiment. Furthermore, the culture apparatus 1 may also be configured to include a second sensor 13 that measures the concentration of the dissolved gas to be adjusted in the culture solution 3, and the control device 12 is electrically connected to the sensor 11 and the second sensor 13 and controls the three-way valve serving as the flow rate regulator 10 in accordance with the measurement results from the sensor 11 and the second sensor 13. The second sensor 13 may be configured to be provided upstream of the three-way valve in the reactor 2a, as in this embodiment. The sensor 11 and the second sensor 13 may be configured to measure the pH or temperature, or both, of the culture solution 3 in addition to the concentration of the dissolved gas to be adjusted in the culture solution 3. The control device 12 may be configured to control the three-way valve in accordance with the in vivo metabolism or circadian rhythm of the culture target, the amount of light, or the like, in addition to the measurement results from the sensor 11 (and the second sensor 13). The control device 12 is not limited to control in accordance with the measurement results from the sensor 11 (and the second sensor 13), and may be configured to perform control in accordance with, for example, a preset time schedule.

[0023] The gas concentration adjusting unit 6 includes a (first) check valve 6d that allows the culture solution 3 to flow from the branching portion 2b to the confluence portion 2c in the branching path 5 while suppressing flow in the reverse direction. According to the above configuration, the check valve 6d allows the culture solution 3, whose concentration has been adjusted, to flow stably to the confluence portion 2c, thereby enabling stable adjustment of the concentration of the dissolved gas to be adjusted in the culture solution 3. In this embodiment, the check valve 6d is provided between the branching portion 2b and the gas-permeable member 6a. To enhance the effect of stabilizing the concentration adjustment, the gas concentration adjusting unit 6 may also include a second check valve 6e, as in this embodiment, that is provided between the confluence portion 2c and the gas-permeable member 6a in the branching path 5 while allowing the culture solution 3 to flow from the branching portion 2b to the confluence portion 2c while suppressing flow in the reverse direction. Furthermore, the gas concentration adjuster 6 may be configured to have the second check valve 6e without the check valve 6d, or may be configured to have neither the check valve 6d nor the second check valve 6e.

[0024] The branching portion 2b and the confluence portion 2c are located inside the reactor 2a. According to the above configuration, the concentration of the dissolved gas to be adjusted in the culture solution 3, which has changed due to the activity of the culture subject in the reactor 2a, can be efficiently adjusted.

[0025] The culture subject may be a microorganism that requires dissolved gases such as carbon dioxide for cultivation, such as hydrogen bacteria (Hydrogenobacter thermophilus, Hydrogenophilus thermoluteolus, etc.). It may also be (micro)algae (cyanobacteria, green algae, etc.) that require dissolved carbon dioxide to be consumed by photosynthesis for cultivation. As the culture solution 3, for example, a liquid medium containing a nitrogen source, a phosphorus source, trace elements, a carbon source, etc. can be used.

[0026] The culture target is cultured in the reactor 2a by consuming the necessary dissolved gases while circulating within the circulation path 2 in a state contained in the culture solution 3. In this embodiment, the reactor 2a has a long flow path that constitutes part of the circulation path 2. The reactor 2a may be configured to have a pipe that forms the flow path inside, as in this embodiment. Furthermore, as in this embodiment, the reactor 2a is preferably a closed system reactor in which the circulation path 2 within the reactor 2a is closed from the outside. The circulation path 2 may be configured to have a tank 2d that stores the culture solution 3, as in this embodiment.

[0027] When the culture target is a microorganism that requires dissolved gas for cultivation, an insufficient concentration of the necessary dissolved gas in the culture solution 3 will result in poor growth of the culture target.

[0028] When the culture subject is a photosynthetic organism, the reactor 2a is preferably a photobioreactor. That is, the culture subject is cultured by receiving light in the reactor 2a. The culture subject exposed to light in the reactor 2a consumes dissolved carbon dioxide in the culture solution 3 through photosynthesis, thereby increasing the dissolved oxygen in the culture solution 3. An insufficient concentration of dissolved carbon dioxide in the culture solution 3 leads to poor growth of the culture subject. Furthermore, an excessive concentration of dissolved oxygen in the culture solution 3 has an adverse effect on the growth of the culture subject. In this embodiment, the reactor 2a is optically transparent, allowing light from an external light source to pass into the reactor 2a.

[0029] When the culture target is a microorganism that requires carbon dioxide for cultivation or when the culture target is an organism that performs photosynthesis, it is preferable that the supply gas supplied by supply path 8 contains carbon dioxide and the dissolved gas to be adjusted contains carbon dioxide. According to the above configuration, carbon dioxide supplied at an appropriate pressure from supply path 8 to branch path 5 of gas concentration adjuster 6 permeates gas permeable member 6a and is directly dissolved in culture solution 3, so that carbon dioxide can be dissolved efficiently in culture solution 3 without waste.

[0030] When the subject of cultivation is a photosynthetic organism, it is preferable that the supply gas supplied through supply path 8 contains nitrogen and the dissolved gas to be adjusted contains oxygen. According to the above configuration, nitrogen is supplied from supply path 8 to branch path 5 at an appropriate pressure, and the nitrogen and culture solution 3 are brought into contact with each other via gas-permeable member 6a. This allows the dissolved oxygen in culture solution 3 to pass through gas-permeable member 6a and be released into the nitrogen gas, thereby efficiently reducing the dissolved oxygen in culture solution 3.

[0031] When the culture target is a photosynthetic organism, it is more preferable that the supply gas supplied through the supply path 8 contains carbon dioxide and nitrogen, and the dissolved gas to be adjusted contains carbon dioxide and oxygen. In this case, the culture apparatus 1 may be configured to have a (first) gas supply source 7 that supplies one of carbon dioxide and nitrogen, and a second gas supply source 14 that supplies the other of carbon dioxide and nitrogen, as shown in the example of FIG. 5. The supply of the supply gas from the gas supply source 7 may be configured to be supplied via a pressure adjustment valve 8a, as in this example. The supply of the supply gas from the second gas supply source 14 may be configured to be supplied via a second pressure adjustment valve 8b, as in this example. The supply of the supply gas from the gas supply source 7 and the supply of the supply gas from the second gas supply source 14 may be configured to be performed simultaneously, or may be configured to be performed at different times.

[0032] 8, the method of operating the culture apparatus 1 in this embodiment may be configured to perform the following steps in this order: a supply step (see FIG. 1) in which pressurized supply gas supplied from the supply path 8 is brought into contact with the culture solution 3 via the gas-permeable member 6a to adjust the concentration of the dissolved gas to be adjusted in the culture solution 3 in the branch path 5; a discharge step (see FIG. 2) in which the pressure of the supply gas in the branch path 5 is reduced by discharging the pressurized supply gas from the branch path 5; and a confluence step (see FIG. 3) in which the culture solution 3, the concentration of which has been adjusted, is returned from the branch path 5 to the confluence section 2c of the circulation path 2. According to the above configuration, by performing the discharge step between the supply step and the confluence step, it is possible to prevent the culture solution 3 from flowing with excessive force from the branch path 5 to the confluence section 2c due to excessive pressure in the branch path 5 in the confluence step, thereby suppressing damage to the culture target.

[0033] The supply path 8 has a first on-off valve 8c, the discharge path 9 has a second on-off valve 9a, the gas concentration adjuster 6 has a third on-off valve 6f provided between the branching section 2b and the gas permeable member 6a, and a fourth on-off valve 6g provided between the gas permeable member 6a and the confluence section 2c, the supply step is performed by opening the first on-off valve 8c and closing the second on-off valve 9a, the third on-off valve 6f, and the fourth on-off valve 6g, the discharge step is performed by opening the second on-off valve 9a and closing the first on-off valve 8c, the third on-off valve 6f, and the fourth on-off valve 6g, and the confluence step is performed by opening the third on-off valve 6f and the fourth on-off valve 6g and closing the first on-off valve 8c. The confluence step may be performed with the second on-off valve 9a open as in this embodiment, or may be performed with it closed.

[0034] The discharge step may be performed so that the pressure of the supply gas in the branch path 5 is slightly (for example, 10 Pa to 20 Pa) higher than atmospheric pressure. According to the above configuration, in the confluence step, the pressure of the supply gas in the branch path 5 can be used to cause the culture solution 3 to flow with an appropriate force from the branch path 5 to the confluence part 2c. In this case, the confluence step is performed with the second on-off valve 9a closed.

[0035] The gas concentration adjusting unit 6 has a third sensor 6i that measures the concentration of a dissolved gas to be adjusted in the culture solution 3 in a gas concentration adjusting tank 6h (described later). The control device 12 is electrically connected to the third sensor 6i, the first on-off valve 8c, the second on-off valve 9a, the third on-off valve 6f, and the fourth on-off valve 6g, and controls the first on-off valve 8c, the second on-off valve 9a, the third on-off valve 6f, and the fourth on-off valve 6g in accordance with the measurement results of the third sensor 6i. The control device 12 is also electrically connected to the pressure adjusting valve 8a, and controls the pressure adjusting valve 8a in accordance with the measurement results of the third sensor 6i. The control device 12 is not limited to control in accordance with the measurement results of the third sensor 6i, and may be configured to perform control in accordance with, for example, a preset time schedule.

[0036] As shown in the example of FIG. 4, the gas concentration adjustment unit 6 may have a branch path 5 having a (first) gas concentration adjustment tank 6h with a (first) gas permeable member 6a installed therein, which receives supply gas from a supply path 8 and discharges the supply gas to a discharge path 9, a second gas concentration adjustment tank 6j with a second gas permeable member (not shown) installed therein and connected to the branch path 5 in parallel with the gas concentration adjustment tank 6h, a second supply path 15 which supplies supply gas to the second gas concentration adjustment tank 6j, and a second discharge path 16 which discharges the supply gas from the second gas concentration adjustment tank 6j. In this case, the operating method of the culture apparatus 1 may be configured such that a second confluence step (see thick arrow in FIG. 4) is performed in the second gas concentration adjustment tank 6j while a (first) supply step (see hollow arrow in FIG. 4) and a (first) discharge step are performed in the gas concentration adjustment tank 6h, and a second supply step and a second discharge step are performed in the second gas concentration adjustment tank 6j while the (first) confluence step is performed in the gas concentration adjustment tank 6h. The operating method of the culture apparatus 1 may be configured such that the supply step and the second supply step are performed simultaneously, the discharge step and the second discharge step are performed simultaneously, and the confluence step and the second confluence step are performed simultaneously.

[0037] In this example, the second supply path 15 has a fifth on-off valve 15a, the second exhaust path 16 has a sixth on-off valve 16a, the gas concentration adjustment unit 6 has a seventh on-off valve 6k provided between the branching portion 2b and the second gas permeable member, and an eighth on-off valve 6l provided between the second gas permeable member and the confluence portion 2c, the second supply step is performed by opening the fifth on-off valve 15a and closing the sixth on-off valve 16a, the seventh on-off valve 6k, and the eighth on-off valve 6l, the second exhaust step is performed by opening the sixth on-off valve 16a and closing the fifth on-off valve 15a, the seventh on-off valve 6k, and the eighth on-off valve 6l, and the second confluence step is performed by opening the seventh on-off valve 6k and the eighth on-off valve 6l, and closing the fifth on-off valve 15a.

[0038] In this example, the gas concentration adjuster 6 has a fourth sensor 6m that measures the concentration of the dissolved gas to be adjusted in the culture solution 3 in the second gas concentration adjustment tank 6j, and the control device 12 is electrically connected to the fourth sensor 6m, the fifth on-off valve 15a, the sixth on-off valve 16a, the seventh on-off valve 6k, and the eighth on-off valve 6l, and controls the fifth on-off valve 15a, the sixth on-off valve 16a, the seventh on-off valve 6k, and the eighth on-off valve 6l in accordance with the measurement result by the fourth sensor 6m. Also in this example, the control device 12 is electrically connected to the pressure adjustment valve 8a, and controls the pressure adjustment valve 8a in accordance with the measurement result by the third sensor 6i, the fourth sensor 6m, or both.

[0039] The filter 6b is formed of, for example, a membranous porous member (porous membrane) having a large number of micropores that prevent the passage of the culture subject and allow the passage of the culture solution 3. The porous member may be hydrophobic and may be formed of, for example, a material such as polytetrafluoroethylene, polypropylene, or polyethylene.

[0040] The gas-permeable member 6a has a liquid-side surface that contacts the culture solution 3 and a gas-side surface that contacts the supply gas on the opposite side of the liquid-side surface, and prevents the passage of the liquid (culture solution 3) while allowing the passage of the gas (the dissolved gas to be adjusted). The gas-permeable member 6a preferably has a permeable membrane having a liquid-side surface and a gas-side surface. That is, the gas-permeable member 6a has a membrane contactor 6a1 (porous membrane or homogeneous membrane) having a liquid-side surface and a gas-side surface. The porous membrane has pores of, for example, several tens of angstroms, which are larger than the molecular size of the dissolved gas to be adjusted, thereby allowing the molecules of the dissolved gas to pass through. The homogeneous membrane is formed, for example, from silicon, and allows the molecules of the dissolved gas to pass through by generating gaps of, for example, several angstroms, which are larger than the molecular size of the dissolved gas to be adjusted due to the thermal motion of polymer chains. The membrane contactor 6a1 may be configured to have a tubular shape with an inner circumferential surface formed by a liquid side surface or a gas side surface (for example, a hollow fiber membrane contactor as shown in FIG. 7), or may be configured to have a flat or curved plate shape. The gas permeable member 6a may be configured by a membrane contactor group 6a2 which is an assembly of the membrane contactors 6a1.

[0041] 6, the gas concentration adjusting tank 6h is a sealed container having a liquid-side internal space facing the liquid-side surface of the gas-permeable member 6a disposed therein, a gas-side internal space facing the gas-side surface of the gas-permeable member 6a, a liquid inlet 6h1 and a liquid outlet 6h2 communicating with the liquid-side internal space, and a gas inlet 6h3 and a gas outlet 6h4 communicating with the gas-side internal space. The positions of the liquid inlet 6h1, the liquid outlet 6h2, the gas inlet 6h3, and the gas outlet 6h4 on the sealed container are not particularly limited. For example, as in this example, the liquid outlet 6h2 may be provided vertically above the liquid inlet 6h1, and the gas outlet 6h4 may be provided vertically below the gas inlet 6h3. If the supply gas turns into bubbles in the culture solution 3 and the bubbles get mixed into the circulation path 2 in the reactor 2a, there is a risk of damaging the culture object. Therefore, a gas trap space may be provided in the liquid-side internal space to separate bubbles by buoyancy from the culture solution 3 discharged from the liquid outlet 6h2 toward the confluence 2c. In this case, the liquid outlet 6h2 is provided below the vertical upper end of the liquid-side internal space, and the space above the liquid outlet 6h2 can be used as the gas trap space.

[0042] The culture apparatus 1 may include a circulation path 2, a pump 4, a (first) gas concentration adjuster 6 that forms a (first) branch path 5 that branches from a (first) branch point 2b of the circulation path 2 and merges with a (first) confluence point 2c of the circulation path 2, and adjusts the concentration of a (first) dissolved gas to be adjusted in the culture solution 3 in the branch path 5, and a second gas concentration adjuster that forms a second branch path that branches from a second branch point of the circulation path 2 and merges with the second confluence point of the circulation path 2, and adjusts the concentration of a second dissolved gas to be adjusted in the culture solution 3 in the second branch path. The positions of the branch point 2b and the second branch point in the circulation path 2 may be the same or different. The positions of the confluence point 2c and the second confluence point in the circulation path 2 may be the same or different. The dissolved gas to be adjusted and the second dissolved gas to be adjusted may be the same or different.

[0043] Although the embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments, and the above-described embodiments can be modified in various ways without departing from the gist of the present invention. [Explanation of symbols]

[0044] 1 Culture device 2 Circulation Route 2a Reactor 2b Branch 2c Confluence 2d tank 3 Culture solution 4. Pump 5 Branching Paths 6 Gas concentration adjustment section 6a Gas permeable member 6a1 Membrane Contactor 6a2 Membrane contactor group 6b Filter 6c Second filter 6d Check valve 6e Second check valve 6f Third shut-off valve 6g 4th shutoff valve 6h Gas concentration adjustment tank 6h1 Liquid inlet 6h2 Liquid outlet 6h3 Gas inlet 6h4 Gas outlet 6i 3rd sensor 6j Second gas concentration adjustment tank 6k 7th shut-off valve 6l No. 8 shut-off valve 6m 4th sensor 7 Gas supply source 8 Supply Channels 8a Pressure regulating valve 8b Second pressure regulating valve 8c First shut-off valve 9. Excretion Route 9a Second shut-off valve 10 Flow rate adjustment section 11 Sensors 12 Control device 13 Second sensor 14 Second gas supply source 15 Second Supply Route 15a 5th shut-off valve 16 Secondary Emission Pathway 16a 6th shut-off valve

Claims

1. a circulation path having a reactor for cultivating a culture target; a pump that circulates the culture solution containing the culture subject through the circulation path; a gas concentration adjusting unit that forms a branch path that branches from the branching portion of the circulation path and joins the joining portion of the circulation path; a supply path for supplying a supply gas from a gas supply source to the branch path; an exhaust path for exhausting the supply gas from the branch path, the gas concentration adjusting unit includes a gas permeable member and a filter that is provided between the branching unit and the gas permeable member and that prevents the culture object from passing through and allows the culture solution to pass through; The gas-permeable member adjusts the concentration of the dissolved gas to be adjusted in the culture solution in the branch path by bringing the supply gas supplied from the supply path into contact with the culture solution via the gas-permeable member.

2. The culture apparatus according to claim 1 , further comprising a flow rate adjusting unit that adjusts the flow rate of the culture solution toward the circulation path side and the flow rate of the culture solution toward the branch path side at the branching section.

3. The culture device according to claim 2 , wherein the flow rate adjusting unit allows the culture medium to flow simultaneously on both the circulation path side and the branch path side.

4. a sensor for measuring the concentration of the dissolved gas to be adjusted in the culture solution; The culture device according to claim 2 , further comprising a control device electrically connected to the sensor and the three-way valve as the flow rate adjusting unit, and controlling the three-way valve in accordance with the measurement result by the sensor.

5. The culture apparatus according to claim 1 , wherein the gas concentration adjusting unit has a check valve that allows the culture solution to flow from the branching portion to the confluence portion in the branching path while suppressing a flow in the reverse direction.

6. The culture device according to claim 1 , wherein the branching portion and the confluence portion are located within the reactor.

7. The culture apparatus according to claim 1 , wherein the supply gas supplied by the supply path contains carbon dioxide, and the dissolved gas to be adjusted contains carbon dioxide.

8. The culture apparatus according to claim 1 , wherein the supply gas supplied by the supply path contains nitrogen, and the dissolved gas to be adjusted contains oxygen.

9. a supply step of adjusting the concentration of the dissolved gas to be adjusted in the culture solution in the branch path by bringing the pressurized supply gas supplied from the supply path into contact with the culture solution via the gas permeable member; a discharge step of discharging the pressurized supply gas from the branch passage to reduce the pressure of the supply gas in the branch passage; a confluence step of returning the culture solution, in which the concentration of the dissolved gas to be adjusted has been adjusted, from the branch path to the confluence part of the circulation path, in this order.

10. the supply path has a first on-off valve; the discharge path has a second on-off valve; the gas concentration adjusting unit includes a third on-off valve provided between the branching portion and the gas permeable member, and a fourth on-off valve provided between the gas permeable member and the junction portion, the supplying step is performed by opening the first on-off valve and closing the second on-off valve, the third on-off valve, and the fourth on-off valve, the discharge step is performed by opening the second on-off valve and closing the first on-off valve, the third on-off valve, and the fourth on-off valve, The method for operating a culture apparatus according to claim 9 , wherein the confluence step is performed by opening the third on-off valve and the fourth on-off valve and closing the first on-off valve.

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