Sampling device and cell culture system

Abstract

The cell culture system (10) is equipped with a culture device (11) having a plurality of reactors (12) and a sampling device (60) for collecting samples of a liquid from the culture device (11). The sampling device (60) is provided in a sample introduction pathway (130) and has a temporary storage unit (136) capable of temporarily storing a sample. The temporary storage unit (136) creates a combined sample of a plurality of samples by causing samples from each of the plurality of reactors (12) to flow in sequentially under the operation of an upstream pump (142) and discharges the combined sample to a sampling pathway (64).

Description

[Technical Field]

[0001] The present invention relates to a sampling device for collecting a sample of a liquid in a culture device for culturing cells, and a cell culture system. [Background technology]

[0002] For example, U.S. Patent No. 9,442,047 discloses a sampling device equipped with a sampling path for collecting a liquid sample from a culture device. The sampling device includes an introduction pump that draws a sample from a sample introduction path connected to the culture device into the sampling path, and a detection unit provided downstream of the sampling path. The detection unit detects the components contained in the sample and the amount (concentration) of the components. Summary of the Invention

[0003] This type of culture device may be configured to include multiple reactors, which are culture containers, in order to improve the efficiency of cell culture. In other words, the culture device can culture cells in each of the multiple reactors by seeding cells and supplying culture medium to each of the reactors.

[0004] However, the culture conditions may differ slightly among multiple reactors due to factors such as slight differences in the circulation of the culture medium, etc. Therefore, when a system for detecting samples is constructed by connecting a sampling device to a culture device, even if samples flowing out from an unspecified reactor are detected, the culture conditions of each reactor cannot be accurately determined.

[0005] On the other hand, if the sampling device is configured to detect samples from each of a plurality of reactors, the number of samplings and the amount of samples will increase significantly, resulting in a decrease in sampling efficiency.

[0006] The present invention has been made in consideration of the above circumstances, and has an object to provide a sampling device and a cell culture system that can more efficiently detect samples from each of a plurality of reactors.

[0007] In order to achieve the above-mentioned object, a first aspect of the present invention is a sampling device for collecting a liquid sample from a culture device having a plurality of reactors for culturing cells based on the flow of culture medium, comprising: a sampling path through which the sample flows; a detection unit provided in the sampling path; a sample introduction path connecting the sampling path upstream of the detection unit and the culture device; a pump for circulating the sample through the sample introduction path; and a control unit for operating the pump.The sampling path also has a temporary storage unit that is provided in the sample introduction path and can temporarily store the sample, and the temporary storage unit combines the plurality of samples into a combined sample by sequentially flowing the samples from each of the plurality of reactors into the temporary storage unit under the operation of the pump, and then discharges the combined sample into the sampling path.

[0008] In order to achieve the above-mentioned object, a second aspect of the present invention is a cell culture system that collects a liquid sample from a culture unit having a plurality of reactors that culture cells based on the flow of culture medium, wherein the culture unit sequentially supplies culture medium to the plurality of reactors and comprises a sampling path through which the sample flows, a detection unit provided in the sampling path, a sample introduction path that connects the sampling path upstream of the detection unit to the culture unit, a pump that circulates the sample through the sample introduction path, and a control unit that operates the pump, and the sample introduction path has a temporary storage unit that is provided in the sample introduction path and can temporarily store the sample, and the temporary storage unit combines the plurality of samples into a combined sample by sequentially flowing the samples from each of the plurality of reactors into the temporary storage unit under the operation of the pump, and then discharges the combined sample into the sampling path.

[0009] The above-described sampling device and cell culture system can detect samples from multiple reactors more efficiently. [Brief explanation of the drawings]

[0010] [Figure 1]1 is a perspective view schematically showing the overall configuration of a cell culture system to which a sampling device according to one embodiment of the present invention is applied. [Figure 2] FIG. 1 is an explanatory diagram schematically showing the path of a medium during cell culture. [Figure 3] FIG. 1 is an explanatory diagram schematically showing the paths through which samples flow out from multiple reactors. [Figure 4] FIG. 2 is an explanatory diagram illustrating a schematic path of a sampling device. [Figure 5] 1 is a flowchart illustrating a sampling method of the sampling device. [Figure 6] 1 is a flowchart showing a sampling process. [Figure 7] FIG. 10 is an explanatory diagram showing the operation of a temporary storage step. [Figure 8] FIG. 10 is an explanatory diagram showing the operation of the combined sample outflow process. [Figure 9] FIG. 10 is an explanatory diagram schematically showing a path of a sampling device according to a first modified example. [Figure 10] Fig. 10A is a flowchart showing the sampling process of the sampling device according to Modification 1. Fig. 10B is a flowchart showing the culturing process of the culturing device. [Figure 11] FIG. 10 is an explanatory diagram schematically showing a path of a sampling device according to a second modified example. DETAILED DESCRIPTION OF THE INVENTION

[0011] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to preferred embodiments thereof and the accompanying drawings.

[0012] As shown in Fig. 1, a sampling device 60 according to one embodiment of the present invention is applied to a cell culture system 10 that cultures living cells in regenerative medicine. The sampling device 60 samples the culture medium during cell culture in the cell culture system 10 to measure the state of the culture medium. For example, the cell culture system 10 continues cell culture for a long period of time by supplying culture medium and oxygen to a reactor 12, which is a cell culture container, while discharging lactic acid, carbon dioxide, and the like (including unused culture medium and oxygen) generated during cell culture from the reactor 12.

[0013] The biological cells are not particularly limited, but examples thereof include cells contained in blood (T cells, etc.) and stem cells (ES cells, iPS cells, mesenchymal stem cells, etc.). The medium may also be selected appropriately depending on the biological cells, and examples thereof include those prepared by adding various amino acids, vitamins, serum, etc. to a balanced salt solution (BSS) as a base solution.

[0014] The cell culture system 10 includes a culture device 11 (culture unit) in which multiple reactors 12 are set and cells are actually cultured, and a sampling device 60 (sampling unit) that collects liquid samples from the culture device 11 during culture. That is, the cell culture system 10 is configured to circulate a culture medium through each of the multiple reactors 12 and culture cells in each reactor 12, thereby obtaining several times the number of cells compared to culturing in a single reactor 12 without significantly changing the culture period. Note that while FIG. 1 illustrates the culture device 11 with five reactors 12, the number of reactors 12 provided in the culture device 11 is not particularly limited. Furthermore, the cell culture system 10 may be configured such that multiple culture devices 11 are connected to one sampling device 60. Furthermore, although the present embodiment illustrates a cell culture system 10 in which the culture unit and the sampling unit are configured separately, the cell culture system 10 may also be an apparatus in which the culture unit and the sampling unit are integrated (united).

[0015] The culture device 11 has a culture medium storage section 14 that stores culture medium, a circulation path 16 that is provided between the reactor 12 and the culture medium storage section 14, a plurality of medical bags 18 that are connected to the circulation path 16, and a waste liquid section 20 that stores liquid discharged from the circulation path 16.

[0016] A hard tank capable of storing a large amount of culture medium is applied as the culture medium storage unit 14. The distribution path 16 is composed of a plurality of tubes 22, which are connected to the plurality of reactors 12, the culture medium storage unit 14, the plurality of medical bags 18, and the waste liquid unit 20, respectively.

[0017] The multiple medical bags 18 include, for example, a cell fluid bag 18A that stores a fluid containing cells (cell fluid), a cleaning fluid bag 18B that stores a cleaning fluid, a detachment fluid bag 18C that stores a detachment fluid, and a recovery bag (not shown) that recovers the cultured cells. The cleaning fluid is a liquid used when priming the reactor 12 and the distribution channel 16. Examples of this cleaning fluid include buffer solutions such as PBS (Phosphate Buffered Salts) and TBS (Tris-Buffered Saline), or physiological saline. The detachment fluid is a liquid that detaches cells cultured by a culture process. Examples of the detachment fluid that can be used include trypsin and EDTA solution.

[0018] When constructing the cell culture system 10, the flow path 16 is set to pass through a flow path control mechanism 24 of the culture device 11. The flow path control mechanism 24 has a housing 26 that houses a part of the flow path 16. The flow path control mechanism 24 also includes, in the housing 26, a clamp 28 that opens and closes a predetermined tube 22, a pump 30 that circulates the liquid in the tube 22, and a control circuit 32 that controls the operation of the clamp 28 and the pump 30 (see FIG. 2).

[0019] The reactors 12 are housed in a housing 26 of the flow path control mechanism 24. The reactor 12 includes a plurality of hollow fibers 34 (e.g., 10,000 or more) and a case 36 that houses the hollow fibers 34. Each hollow fiber 34 has a lumen (not shown), and cells are seeded on the inner circumferential surface that constitutes the lumen. Each hollow fiber 34 also has a plurality of pores (not shown) that communicate between the outside and the lumen. Each pore is impermeable to cells and proteins but permeable to solutions and low-molecular-weight substances. A culture medium or the like is supplied to the cells seeded on the inner circumferential surface of the hollow fibers 34 via the lumen or the pores. Hereinafter, a configuration in which a liquid is circulated mainly through the lumen of the hollow fibers 34 is also referred to as an IC (intra capillary), and a configuration in which a liquid is circulated mainly outside the hollow fibers 34 is also referred to as an EC (extra capillary).

[0020] The case 36 is provided with a first IC terminal 36a and a second IC terminal 36b that communicate with the inner cavity of the hollow fibers 34, and a first EC terminal 36c and a second EC terminal 36d that communicate with the space outside the hollow fibers 34 within the case 36, and a tube 22 is connected to each terminal.

[0021] 2, the configuration of the flow path 16 between one reactor 12 and the culture medium reservoir 14, and the flow path control mechanism 24 will be specifically described. The flow path 16 has a culture medium delivery route 40 connected to the culture medium reservoir 14, and an IC route 42 (internal route) and an EC route 44 (external route) branching off from the culture medium delivery route 40. The IC route 42 is a route for supplying liquid to the lumen of the hollow fibers 34. The EC route 44 is a route for supplying liquid to the inside of the case 36 outside the hollow fibers 34.

[0022] The IC route 42 has an IC circulation circuit 42a capable of circulating liquid between the reactor 12 and the IC route 42, and an IC supply circuit 42b capable of distributing liquid from the culture medium delivery route 40 to the IC circulation circuit 42a. The IC circulation circuit 42a is connected to the first IC terminal 36a and the second IC terminal 36b of the reactor 12, and is equipped with an IC circulation pump 30a that circulates liquid through the lumen of the hollow fibers 34. An IC waste liquid circuit 46 that discharges the culture medium to the waste liquid section 20 is connected to the IC circulation circuit 42a downstream of the reactor 12. Meanwhile, the IC supply circuit 42b is equipped with an IC supply pump 30b that circulates liquid from the culture medium delivery route 40 to the IC circulation circuit 42a.

[0023] Meanwhile, the EC route 44 includes an EC circulation circuit 44a capable of circulating liquid between the reactor 12 and the EC route 44, and an EC supply circuit 44b capable of circulating liquid from the culture medium delivery route 40 to the EC circulation circuit 44a. The EC circulation circuit 44a is connected to the first EC terminal 36c and the second EC terminal 36d of the reactor 12, and includes an EC circulation pump 30c that circulates liquid around the outside of the hollow fibers 34. A gas exchanger 52 is provided upstream of the reactor 12 in the EC circulation circuit 44a. The gas exchanger 52 discharges carbon dioxide contained in the culture medium while mixing predetermined gas components (e.g., nitrogen (N2): 75%, oxygen (O2): 20%, carbon dioxide (CO2): 5%) into the culture medium. An EC waste liquid circuit 48 that discharges the culture medium to the waste liquid section 20 is connected downstream of the reactor 12 in the EC circulation circuit 44a. The EC supply circuit 44b is provided with an EC supply pump 30d that circulates the liquid from the culture medium delivery route 40 to the EC circulation circuit 44a.

[0024] Although not shown, in addition to the culture medium reservoir 14, a plurality of medical bags 18 (cell solution bag 18A, cleaning solution bag 18B, detachment solution bag 18C) are connected to the IC supply circuit 42b upstream of the IC supply pump 30b or the EC supply circuit 44b upstream of the EC supply pump 30d via a plurality of tubes 22. These medical bags 18 may be replaced with collection bags or the like using a sterile joining device that sterilizes and joins the bags depending on the application.

[0025] The sampling device 60 is connected to the EC circulation circuit 44a of the culture device 11 at a position near the downstream side (second EC terminal 36d) of the reactor 12 (between the reactor 12 and the EC waste liquid circuit 48). For this reason, one end of a sample outflow path 54, through which a culture medium, which is a liquid sample, flows out, is connected to the EC circulation circuit 44a. A culture device-side connector 56 is provided at the other end of the sample outflow path 54. The culture device-side connector 56 is configured to be mutually connectable with the sampling device-side connector 132 of the sampling device 60. The sampling device 60 may also be connected to the downstream side (second IC terminal 36b) of the reactor 12 of the IC circulation circuit 42a via the sample outflow path 54.

[0026] 2 and 3, the cell culture system 10 includes a plurality of IC circulation circuits 42a and EC circulation circuits 44a corresponding to a plurality (five) of reactors 12. That is, at a branch point X between the IC supply pump 30b and the IC circulation circuit 42a, and at a branch point Y between the EC supply pump 30d and the EC circulation circuit 44a, other IC circulation circuits 42a and EC circulation circuits 44a (not shown) that circulate liquid to other reactors 12 are connected in parallel. The EC supply circuits 44b between the branch point Y and each EC circulation circuit 44a are each provided with a supply clamp 29 that switches on and off the supply of culture medium to each EC circulation circuit 44a.

[0027] Hereinafter, the five reactors 12 in FIG. 3 are referred to as reactor 12A to reactor 12E from top to bottom. The supply clamps 29 provided in each EC supply circuit 44b are referred to as supply clamps 29A to 29E corresponding to the reactors 12A to 12E. The culture device 11 opens one of the supply clamps 29A to 29E and closes the other four while rotating the EC supply pump 30d. This causes the culture medium to be supplied to the EC circulation circuit 44a whose supply clamp 29 is open, and the culture medium circulates in the EC circulation circuit 44a and flows through the reactor 12. Although not shown, the culture device 11 has multiple IC circulation circuits 42a corresponding to the reactors 12A to 12E. Each IC supply circuit 42b connected to each of the multiple IC circulation circuits 42a is also provided with a supply clamp (not shown) for selectively distributing the culture medium.

[0028] The sample outflow path 54 branches at a branch point Z as a base point and connects to each of the EC circulation circuits 44a in order to connect to each of the multiple reactors 12. A sterile filter 58 is provided in the sample outflow path 54 from the branch point Z to the culture device side connector 56. The sterile filter 58 maintains the sterility of the culture medium flowing on the culture device 11 side (EC circulation circuit 44a side).

[0029] Next, the configuration of the sampling device 60 will be described with reference to Figure 4. The sampling device 60 collects samples of culture medium from one or more culture devices 11 and detects the components contained in the sample and the amounts (concentrations) of the components. The sampling device 60 includes a sampling kit 62 having a sampling path 64 through which the sample is collected, multiple mechanical units 66 to which the sampling kit 62 is detachably set, and a controller 68 that controls the operation of the multiple mechanical units 66. The sampling kit 62 is a disposable item, and the multiple mechanical units 66 are reusable items.

[0030] In addition to the sampling path 64, the sampling kit 62 includes a cleaning liquid storage section 70, a standard liquid storage section 72, a waste liquid storage section 74, and a detection section 75 (first detection section 76, second detection section 80). The sampling path 64 is composed of a flexible tube of an appropriate diameter to allow the sample to flow through. The cleaning liquid storage section 70 is connected to a branch point 65 to which one end of the sampling path 64 is connected via a cleaning liquid branch path 71, and the standard liquid storage section 72 is connected to this branch point 65 via a standard liquid branch path 73. The other end of the sampling path 64 is connected to the waste liquid storage section 74.

[0031] The cleaning liquid storage section 70 and the standard liquid storage section 72 are formed in a bag shape (medical bag) from a soft resin material such as polyvinyl chloride or polyolefin. However, there are no particular limitations on the cleaning liquid storage section 70 and the standard liquid storage section 72 as long as they are capable of storing liquid. The waste liquid storage section 74 shares a tank with the waste liquid section 20 of the culture device 11, but is not limited to this and a medical bag or the like may be used.

[0032] A cleaning liquid is stored in the cleaning liquid storage section 70. The cleaning liquid is not particularly limited, and for example, the buffer solution, physiological saline, etc. listed as the cleaning liquid for the cleaning liquid bag 18B of the culture device 11 may be used appropriately.

[0033] A standard solution is stored in the standard solution storage section 72. The standard solution is a liquid for calibrating the first detection section 76 and the second detection section 80, and is a liquid in which the pH value, glucose value (glucose concentration), and lactic acid value (lactic acid concentration) are set to specified values.

[0034] The first detection unit 76 and the second detection unit 80 are provided in series and spaced apart from each other at a position midway along the sampling path 64. Note that the detection unit 75 is not limited to a structure in which the first detection unit 76 and the second detection unit 80 are separated, but may be a structure in which the first detection unit 76 and the second detection unit 80 are integrated, or may be a structure in which the first detection unit 76 and the second detection unit 80 are separated into three or more units.

[0035] The first detection unit 76 is a cylindrical member having a plurality of first element units 78 that come into contact with the sample (wetted) in the flow path of the sampling path 64. For example, the plurality of first element units 78 include a pH tip 78a for measuring the pH of the sample, an O2 tip 78b for measuring the O2 concentration in the sample, and a CO2 tip 78c for measuring the CO2 concentration in the sample. The pH tip 78a is a tip for measuring H + , O.H. - The O2 tip 78b changes color in response to O2, and the CO2 tip 78c changes color in response to CO2.

[0036] The second detection unit 80 is a tubular member having a plurality of second element units 82 that come into contact with the sample (wetted) in the flow path of the sampling path 64, and is provided downstream (in the waste liquid storage unit 74) of the first detection unit 76. For example, the plurality of second element units 82 are biosensors that react an enzyme with the flowing sample and detect changes in current, etc. The plurality of second element units 82 include a glucose chip 82a that measures the glucose concentration in the sample and a lactic acid chip 82b that measures the lactic acid concentration in the sample. The glucose chip 82a is electrically connected to a glucose terminal 83a that protrudes outside the tubular member. The lactic acid chip 82b is electrically connected to a lactic acid terminal 83b that protrudes outside the tubular member.

[0037] The sampling kit 62 also includes a connection site 84 between the branch point 65 of the sampling path 64 and the first detection unit 76, to which one or more sample introduction paths 130 (described below) can be connected. The connection site 84 is, for example, a member formed by integrally molding multiple branch ports each having a valve (not shown) that closes when the sample introduction path 130 is not attached and opens when the sample introduction path 130 is attached (in FIG. 4, the connection site 84 is conveniently indicated by the area surrounded by a two-dot chain line). Alternatively, the connection site 84 can be a port to which the sample introduction path 130 can be connected while ensuring the sterility of the sampling path 64.

[0038] As shown in Figure 4, a portion of the sampling kit 62 described above is set in a main mechanism 90, which is one of the multiple mechanism sections 66. The main mechanism section 90 includes a main mechanism section pump 92 and multiple clamps 94 that open and close the flow paths in each path (tube) within a housing 91. Although not shown, it is preferable that a controller 68 that controls the sampling device 60 is also provided in the main mechanism section 90. By setting the sampling kit 62 in the main mechanism section 90, a main unit 96 of the sampling device 60 is constructed.

[0039] The main mechanism side pump 92 is provided with a sampling path 64 extending between the branch point 65 and the connection part 84. The main mechanism side pump 92 has a circular wrapped portion around which the sampling path 64 can be wrapped, and rotates to squeeze the wrapped sampling path 64 (tube), thereby circulating the internal fluid (liquid, air, etc.).

[0040] The multiple clamps 94 include a cleaning liquid clamp 94a that opens and closes the cleaning liquid branch path 71, a standard liquid clamp 94b that opens and closes the standard liquid branch path 73, and a waste liquid clamp 94c that opens and closes the sampling path 64 between the second detection section 80 and the waste liquid storage section 74.

[0041] Furthermore, the first detection unit 76 of the sampling kit 62 is set in a first measurement device 110, which is one of the multiple mechanism units 66, to form a first sensor unit 111. The first measurement device 110 has a holder 112 that houses the first detection unit 76, and a measurement main body unit 114 that is fixed to the holder 112 and optically measures the multiple first element units 78.

[0042] The measurement main body 114 has a PH detector 116a, an O2 detector 116b, and a CO2 detector 116c that face the PH chip 78a, the O2 chip 78b, and the CO2 chip 78c when the first detection unit 76 is held in the holder 112. Under the control of the controller 68, the measurement main body 114 emits measurement light of a wavelength according to the characteristics of each first element unit 78, receives excitation light resulting from excitation of each first element unit 78, and transmits a detection signal to the controller 68.

[0043] Furthermore, the second detection unit 80 of the sampling kit 62 is configured as a second sensor unit 121 by being set in a second measuring device 120, which is one of the multiple mechanical units 66. The second measuring device 120 has a case 122 that can house the second detection unit 80, and an enzyme detector (not shown) that is electrically connected to the glucose terminal 83a and the lactic acid terminal 83b. The enzyme detector detects current values ​​from each of the glucose tip 82a and the lactic acid tip 82b, and transmits a detection signal based on the current values ​​to the controller 68.

[0044] A sample introduction path 130 is connected to the connection site 84 of the sampling kit 62 (sampling path 64) in order to introduce a sample to be measured by the first sensor unit 111 and the second sensor unit 121. Similar to the sampling path 64, the sample introduction path 130 is made of a flexible tube having an appropriate thickness to allow the sample to flow through.

[0045] The sample introduction path 130 has at one end a sampling device side connector 132 for connection to the above-mentioned culture device side connector 56 (see also Figures 2 and 3). At the other end of the sample introduction path 130, a detachable plug (not shown) is provided at the connection site 84. Hereinafter, the point where the sample introduction path 130 is connected to the sampling path 64 will be referred to as the connection point 134.

[0046] A temporary storage section 136 is provided in the sample introduction path 130 between the sampling device side connector 132 and the plug (connection point 134). The temporary storage section 136 temporarily stores the sample that has flowed out from the culture device 11, and then causes the sample to flow out toward the sampling path 64. For example, the temporary storage section 136 is implemented as a medical bag that is softer than the sample introduction path 130, and is hung from a stand 98 fixed on the housing 91 of the main unit 96. The temporary storage section 136 may also be implemented as a hard container.

[0047] The sample introduction path 130 includes an upstream line 137 provided between the sampling device side connector 132 and the temporary storage section 136, and a downstream line 138 provided between the temporary storage section 136 and the plug. When the temporary storage section 136 is suspended from the stand 98, the upstream line 137 and the downstream line 138 are connected to the lower side of the temporary storage section 136 in the direction of gravity.

[0048] A portion of the sample introduction path 130 is detachably set in an introduction mechanism 140, which is one of the multiple mechanism parts 66, to form an introduction unit 148 of the sampling device 60. The introduction mechanism part 140 includes an upstream pump 142, an introduction pump 144, and a downstream clamp 146 in a housing 141. Furthermore, the introduction mechanism part 140 may be configured to include a sensor (not shown) that detects pressure and air bubbles within the flow path of the sample introduction path 130.

[0049] The introduction unit 148 allows a part of the sample introduction path 130, the upstream pump 142, the introduction pump 144, and the downstream clamp 146 to be handled together. A short portion of the sample introduction path 130 (downstream line 138) extending from the introduction unit 148 is connected to the connection site 84 on the housing 91.

[0050] The upstream pump 142 is disposed in the upstream line 137 (i.e., between the culture device 11 and the temporary storage section 136) in the introduction unit 148. The introduction pump 144 and the downstream clamp 146 are disposed in the downstream line 138 (i.e., between the sampling path 64 and the temporary storage section 136) in the introduction unit 148. The upstream pump 142 and the introduction pump 144 have circular wound portions around which the sample introduction path 130 can be wound, and rotate in a manner that squeezes the wrapped-around sample introduction path 130 (tube), thereby circulating the internal fluid. The downstream clamp 146 opens and closes the downstream line 138, thereby switching between allowing and stopping the outflow of the sample from the temporary storage section 136 to the sampling path 64.

[0051] The controller 68 (control unit) is a computer having one or more processors, a memory, an input / output interface, and electronic circuits (not shown). The controller 68 controls the entire sampling device 60 by having the processor execute a program stored in the memory. In this embodiment, the controller 68 is configured to be able to communicate information with the control circuit 32 of the culture device 11, and controls the culture device 11 and the sampling device 60 in conjunction with each other. The controller 68 may also be a control device integrated with the control circuit 32 of the culture device 11.

[0052] The sampling device 60 according to this embodiment is basically configured as described above, and the sampling method of the sampling device 60 will be described below with reference to Fig. 5. The sampling method sequentially performs a preparation step, a priming step, a sampling step, a cleaning step, and a calibration step.

[0053] First, in the preparation step (step S1), as shown in Fig. 4, the user of the cell culture system 10 sets (attaches) the sampling kit 62 on the main mechanism 90 to form the main unit 96. After that, the user sets the first detection unit 76 exposed from the housing 91 on the first measurement device 110 to form the first sensor unit 111, and also sets the second detection unit 80, which is also exposed, on the second measurement device 120 to form the second sensor unit 121. The first sensor unit 111 and the second sensor unit 121 are hung on a stand 98.

[0054] Furthermore, the user sets the sample introduction path 130 in the introduction mechanism part 140 to form the introduction unit 148. After that, the user connects the sampling device side connector 132 of the sample introduction path 130 exposed from the introduction unit 148 to the culture device side connector 56, and also connects the plug of the sample introduction path 130 to the connection part 84.

[0055] Subsequently, in the priming step (step S2 in FIG. 5), the controller 68 opens the cleaning liquid clamp 94a and the waste liquid clamp 94c, while closing the standard liquid clamp 94b, and rotates the main mechanism pump 92. This causes the cleaning liquid in the cleaning liquid storage unit 70 to pass through the first detection unit 76 and the second detection unit 80 and be discharged into the waste liquid storage unit 74.

[0056] Next, in the sampling step (step S3 in FIG. 5), the controller 68 guides the sample from the culture device 11 to the detection unit 75. At this time, as shown in FIG. 6, the controller 68 sequentially carries out a temporary storage step and a combined sample outflow step.

[0057] The temporary storage step is a step of collectively storing the samples flowing out from the reactors 12A to 12E in the temporary storage unit 136. In this embodiment, the controller 68 acquires information on the rotation of the EC supply pump 30d of the culture device 11 and the opening of the supply clamps 29A to 29E, and rotates the upstream pump 142 while the supply clamps 29A to 29E are open. The upstream pump 142 operates at the same rotation speed and for the same time (predetermined period) while the supply clamps 29A to 29E are open. As a result, the temporary storage unit 136 stores the same amount of each sample from the reactors 12A to 12E.

[0058] Specifically, after the start of the temporary storage step, the controller 68 acquires information on the rotation of the EC supply pump 30d and the opening of the supply clamp 29A from the control circuit 32 of the culture device 11 (step S3-1). Thereafter, the controller 68 rotates the upstream pump 142 for a predetermined period while keeping the downstream clamp 146 closed (step S3-2). At this time, the controller 68 also stops the operation of the introduction pump 144 (the same applies hereinafter until the end of the temporary storage step). As a result, the sample flowing out of the reactor 12A is stored in the temporary storage section 136 via the upstream line 137, as shown in FIG. 7 .

[0059] Controller 68 rotates upstream pump 142 to circulate the sample at a flow rate of, for example, 10 mL / min. The predetermined period for operating upstream pump 142 is set to a range of approximately 5 to 15 seconds, depending on the open period of supply clamp 29A. As a result, for example, when upstream pump 142 is rotated for 6 seconds, 1 mL of the sample in reactor 12A is stored in temporary storage section 136.

[0060] Next, the controller 68 acquires information on the rotation of the EC supply pump 30d and the opening of the supply clamp 29B from the control circuit 32 of the culture device 11 (step S3-3). After that, the controller 68 rotates the upstream pump 142 for a predetermined period (the same period as the sample acquisition period for the reactor 12A) while keeping the downstream clamp 146 closed (step S3-4). As a result, the temporary storage unit 136 stores the same amount of sample from the reactor 12B as the amount of sample stored in the reactor 12A.

[0061] Next, the controller 68 acquires information on the rotation of the EC supply pump 30d and the opening of the supply clamp 29C from the control circuit 32 of the culture device 11 (step S3-5). After that, the controller 68 rotates the upstream pump 142 for a predetermined period (the same period as the sample acquisition period for the reactor 12A) while keeping the downstream clamp 146 closed (step S3-6). As a result, the temporary storage unit 136 stores the same amount of sample from the reactor 12C as the amount of sample stored in the reactor 12A.

[0062] Next, the controller 68 acquires information on the rotation of the EC supply pump 30d and the opening of the supply clamp 29D from the control circuit 32 of the culture device 11 (step S3-7). After that, the controller 68 rotates the upstream pump 142 for a predetermined period (the same period as the sample acquisition period for the reactor 12A) while keeping the downstream clamp 146 closed (step S3-8). As a result, the temporary storage unit 136 stores the same amount of sample from the reactor 12D as the amount of sample stored in the reactor 12A.

[0063] Next, the controller 68 acquires information on the rotation of the EC supply pump 30d and the opening of the supply clamp 29E from the control circuit 32 of the culture device 11 (step S3-9). After that, the controller 68 rotates the upstream pump 142 for a predetermined period (the same period as the sample acquisition period of the reactor 12A) while keeping the downstream clamp 146 closed (step S3-10). As a result, the temporary storage unit 136 stores the same amount of sample from the reactor 12E as the amount of sample stored in the reactor 12A.

[0064] By carrying out the above steps S3-1 to S3-10, a combined sample obtained by combining the samples from each of the reactors 12A to 12E is stored in the temporary storage unit 136. Since the same amount of each sample from each of the reactors 12A to 12E flows into the combined sample, it becomes an average of the samples from the entire culture device 11, and can be said to indicate the culture state of the culture device 11.

[0065] After the temporary storage step, the sampling device 60 supplies the combined sample from the temporary storage unit 136 to the sampling path 64 in the next combined sample outflow step. At this time, the controller 68 opens the downstream clamp 146 and rotates the introduction pump 144 (step S3-11). At this time, the controller 68 also stops the operation of the upstream pump 142. As shown in FIG. 8 , the controller 68 closes the cleaning solution clamp 94a and the standard solution clamp 94b, while opening the waste solution clamp 94c and stopping the rotation of the main mechanism pump 92 and the upstream pump 142. As the introduction pump 144 rotates, the combined sample from the temporary storage unit 136 flows out into the downstream line 138 of the sample introduction path 130 at a flow rate of, for example, 10 mL / min. As a result, the combined sample flows from the downstream line 138 into the connection site 84 (connection point 134) of the sampling path 64, passes through the first detection section 76 and the second detection section 80 in order, and is discharged into the waste liquid storage section 74.

[0066] As the combined sample passes, the first element units 78 (pH tip 78a, O2 tip 78b, CO2 tip 78c) of the first detection unit 76 come into contact with the combined sample and change color according to the respective contents of pH, O2, and CO2. The first measuring device 110 performs optical measurement on each of the first element units 78 and transmits the detection results to the controller 68. The controller 68 receives the detection results and performs appropriate processing to display the measurement values ​​(pH value, O2 concentration, CO2 concentration) on the monitor 100 of the main mechanism unit 90.

[0067] Similarly, when the combined sample passes, the multiple second element units 82 (glucose tip 82a, lactic acid tip 82b) of the second detection unit 80 come into contact with the combined sample, and the second measuring device 120 detects current values ​​corresponding to the glucose and lactic acid contents. The second measuring device 120 transmits each detection result to the controller 68. The controller 68 receives the detection results and performs appropriate processing to display the measurement values ​​(glucose concentration, lactic acid concentration) on the monitor 100.

[0068] After the sampling step, the controller 68 determines whether or not cell culture in the culture device 11 has been completed (step S4 in FIG. 5). If cell culture has not been completed (step S4: NO), a cleaning step (step S5 in FIG. 5) is performed. In the cleaning step, the controller 68 supplies the cleaning liquid from the cleaning liquid storage unit 70 to the sampling path 64 to remove the combined sample adhering to the plurality of first element units 78 and the plurality of second element units 82.

[0069] Furthermore, the sampling device 60 performs a calibration process (step S6 in FIG. 5) as necessary. In the calibration process, the controller 68 calibrates the second sensor unit 121 (second measuring device 120) by supplying the standard solution from the standard solution storage section 72 to the sampling path 64. Furthermore, the user calibrates the first measuring device 110 by setting the first measuring device 110 in the calibration device 118 (see FIG. 1).

[0070] When the cleaning step (or calibration step) is completed, the controller 68 returns to step S3 and sequentially performs the subsequent steps. On the other hand, in step S4, if the controller 68 determines that the cell culture is completed (step S4: YES), the operation flow of the sampling device 60 is terminated.

[0071] The sampling device 60 is not limited to the above, and various methods can be adopted. Some other variations of the sampling device 60 will be described below.

[0072] The sampling device 60A according to the first modification is configured to store each sample of each reactor 12 in the temporary storage section 136 without acquiring information on the opening of each of the supply clamps 29A to 29E from the culture device 11. The configuration of the sampling device 60A may be the same as the configuration of the sampling device 60 described above.

[0073] Specifically, as shown in Figure 9, the culture device 11 opens one of the supply clamps 29A-29E and closes the other four clamps for the same period in the order of the reactors 12A-12E while rotating the EC supply pump 30d. This allows the culture device 11 to sequentially and intermittently supply culture medium to each of the EC circulation circuits 44a of the reactors 12A-12E. Furthermore, the culture device 11 opens one of the supply clamps of each of the IC supply circuits 42b while rotating the IC supply pump 30b and closes the other four clamps for the same period in the order of the reactors 12A-12E. This allows the culture device 11 to sequentially and intermittently supply culture medium to each of the EC circulation circuits 44a of the reactors 12A-12E.

[0074] Meanwhile, the controller 68 of the sampling device 60 operates the upstream pump 142 for a period (hereinafter referred to as one cycle period) that is the sum of all the open periods of the supply clamps 29A to 29E. For example, if the open period of each of the supply clamps 29A to 29E of the culture device 11 is 6 seconds, the culture medium is supplied once to each of the reactors 12A to 12E (EC circulation circuit 44a) from the EC supply circuit 44b in 30 seconds. The controller 68 sets the time (30 seconds) for one cycle in which each of the supply clamps 29A to 29E is opened once as one cycle period. The controller 68 then rotates the upstream pump 142 for one cycle period at an appropriate timing when it is deemed necessary to sample the culture medium during the culture process in which the culture medium is supplied and circulated in the culture device 11. The flow rate of each sample caused by the rotation of the upstream pump 142 is set to a value that allows the desired amount of combined sample to be stored in the temporary storage unit 136. As an example, when one cycle period is 30 seconds and 5 mL of the combined sample is stored in temporary storage section 136, upstream pump 142 is rotated so that the flow rate becomes 10 mL / min.

[0075] Here, even if the rotation start timing of upstream pump 142 does not coincide with the release timing of each of supply clamps 29A to 29E, sampling device 60 can collect the same amount of sample from each of reactors 12A to 12E by operating upstream pump 142 for one cycle period. For example, even if the rotation start timing of upstream pump 142 is three seconds later than the release timing of supply clamp 29A, supply clamp 29A is released in the latter half of one cycle period, and each sample can be stored in temporary storage section 136 for the entire open period during which supply clamps 29A to 29E are opened once.

[0076] The sampling device 60A according to the first modified example is basically configured as described above. This sampling device 60A performs the process flow shown in FIG. 10A during the sampling process (step S3 in FIG. 5). That is, when the controller 68 starts the temporary storage process, it rotates the upstream pump 142 for one cycle while keeping the downstream clamp 146 closed (step S3-21). The sampling device 60A is not limited to rotating the upstream pump 142 for only one cycle, but may be configured to rotate the upstream pump 142 two or more times (a or more times: a is a natural number) continuously or intermittently for one cycle. This allows the sampling device 60A to store a sufficient amount of combined sample in the temporary storage section 136, even when the supply rate of the culture medium to each of the reactors 12A-12E is slow, for example.

[0077] 10B, during the temporary storage step, the culture device 11 (control circuit 32) opens supply clamp 29A for a predetermined open period and closes the other supply clamps 29B to 29E for the same period as the open period while rotating the EC supply pump 30d (step S101). This allows the culture medium to be supplied to the EC circulation circuit 44a of the reactor 12A. Similarly, the culture device 11 opens supply clamp 29B for a predetermined open period while rotating the EC supply pump 30d and closes the other supply clamps 29A, 29C to 29E for the same period as the open periods (step S102). Furthermore, the culture device 11 opens supply clamp 29C for a predetermined open period while rotating the EC supply pump 30d and closes the other supply clamps 29A, 29B, 29D, and 29E for the same period as the open periods (step S103). The culture device 11 also opens the supply clamp 29D for a predetermined open period while rotating the EC supply pump 30d, and closes the other supply clamps 29A to 29C and 29E for the same period as the open period (step S104). The culture device 11 also opens the supply clamp 29E for a predetermined open period while rotating the EC supply pump 30d, and closes the other supply clamps 29A to 29D for the same period as the open period (step S105). This sequentially supplies the culture medium to the EC circulation circuits 44a of the reactors 12B to 12E. Finally, the culture device 11 determines whether the culture process is complete (step S106). If the culture process is to be continued (step S106: NO), the process returns to step S101, and the same flow is repeated. If the culture process is to be completed (step S106: YES), the culture device 11 performs the next process (a recovery process of recovering cells from the reactors 12A to 12E).

[0078] 9, the sampling device 60A rotates the upstream pump 142 over one cycle period in which culture medium is supplied to each of the reactors 12A to 12E, thereby storing equal amounts of each sample from each of the reactors 12A to 12E from the culture device 11 in the temporary storage section 136. That is, a combined sample obtained by averaging the individual samples is stored in the temporary storage section 136.

[0079] After the temporary storage step, the controller 68 opens the downstream clamp 146 and rotates the introduction pump 144 (step S3-22). The controller 68 also closes the cleaning solution clamp 94a and the standard solution clamp 94b, while opening the waste solution clamp 94c and stopping the rotation of the main mechanism pump 92 and the upstream pump 142. As a result, the combined sample in the temporary storage section 136 flows into the downstream line 138 of the sample introduction path 130, flows sequentially through the connection part 84 of the sampling path 64, the first detection section 76, and the second detection section 80, and is then discharged into the waste solution storage section 74.

[0080] As described above, by operating the upstream pump 142 for one cycle period, the sampling device 60A can store a combined sample obtained by averaging each sample in the temporary storage section 136 without obtaining information on the opening of each of the supply clamps 29A to 29E from the culture device 11. In particular, since the opening period of the supply clamps 29A to 29E by the culture device 11 does not basically vary, if one cycle period is initially set in the controller 68 of the sampling device 60, periodic sampling can be performed multiple times during the culture process of the culture device 11.

[0081] 11, a sampling device 60B according to the second modification differs from the above-described sampling devices 60 and 60A in that it does not include an upstream pump 142 and a downstream clamp 146, but instead includes an upstream clamp 150 on the upstream line 137. Note that the introduction pump 144 is provided on the downstream line 138, similar to the above-described sampling devices 60 and 60A.

[0082] Furthermore, temporary reservoir 136 is a flexible medical bag that expands as the sample flows in and collapses (flattens) as the sample flows out. One end of upstream line 137 is connected to the lower side, in the direction of gravity, of temporary reservoir 136 held by main unit 96 or introduction unit 148. Conversely, one end of downstream line 138 is connected to the upper side, in the direction of gravity, of temporary reservoir 136 held by main unit 96 or introduction unit 148.

[0083] In the sampling device 60B configured in this manner, during the temporary storage step, the controller 68 rotates the introduction pump 144 while the upstream clamp 150 is open, thereby applying negative pressure to the upstream line 137. In other words, similar to the sampling device 60A, the controller 68 rotates the introduction pump 144 for one cycle period, thereby causing each sample from each of the reactors 12A to 12E to flow into the temporary storage section 136, and the combined sample can be stored.

[0084] Furthermore, in the combined sample discharge step, the sampling device 60B applies negative pressure to the temporary reservoir 136 and the downstream line 138 by rotating the introduction pump 144 while the upstream clamp 150 is closed. As a result, the combined sample in the temporary reservoir 136 flows through the connection part 84 of the sampling path 64, the first detection part 76, and the second detection part 80 in this order, and is discharged into the waste liquid storage part 74.

[0085] In this way, even though the sampling device 60B is configured without the upstream pump 142, it is possible to store each sample of each reactor 12A to 12E in the temporary storage section 136 under the operation of the introduction pump 144. Therefore, the configuration of the introduction unit 148 is further simplified, which reduces costs and makes it easier to handle the introduction unit 148.

[0086] The sampling device 60B can also cause the sample from the culture device 11 to flow into the temporary storage section 136 without relying on the operation of the introduction pump 144. Specifically, the culture device 11 closes the clamps 28 provided on the IC waste liquid circuit 46 and the EC waste liquid circuit 48 (see FIG. 2). This causes the culture medium (waste liquid) from each of the reactors 12A to 12E to flow from the EC circulation circuit 44a to the sample introduction path 130. In other words, the sampling device 60B can store the combined sample in the temporary storage section 136 simply by opening the upstream clamp 150 while the introduction pump 144 is stopped. In this case, because the flow rate of the sample flowing into the sample introduction path 130 depends on the rotation speed of the EC supply pump 30d, the sampling device 60B can appropriately control the opening period of the upstream clamp 150 depending on the rotation speed of the EC supply pump 30d.

[0087] The technical ideas and effects that can be understood from the above-described embodiments will be described below.

[0088] The first aspect of the present invention is a sampling device 60, 60A, 60B for collecting a liquid sample from a culture device 11 having a plurality of reactors 12 for culturing cells based on the flow of culture medium, and is equipped with a sampling path 64 through which the sample flows, a detection unit 75 provided in the sampling path 64, a sample introduction path 130 connecting the sampling path 64 upstream of the detection unit 75 to the culture device 11, pumps (upstream pump 142, introduction pump 144) for circulating the sample through the sample introduction path 130, and a control unit (controller 68) for operating the pump, and a temporary storage unit 136 provided in the sample introduction path 130 for temporarily storing the sample, and the temporary storage unit 136 combines the plurality of samples into a combined sample by sequentially flowing samples from each of the plurality of reactors 12 into it under the operation of the pump, and then discharges the combined sample into the sampling path 64.

[0089] As described above, the sampling devices 60, 60A, 60B can more efficiently detect the samples from the multiple reactors 12 by storing the samples from each of the multiple reactors 12 together in the temporary storage section 136 and discharging the combined sample to the sampling path 64. This allows the sampling devices 60, 60A, 60B to effectively monitor the culture state of the entire culture device 11 (all of the multiple reactors 12) while reducing the number of samplings and the sample amount.

[0090] The culture device 11 also includes a plurality of supply paths (EC supply circuits 44b) for supplying culture medium connected to each of the plurality of reactors 12, as well as a plurality of supply clamps 29 for opening and closing each of the plurality of supply paths, and the control unit (controller 68) operates the pumps (upstream pump 142, introduction pump 144) for a predetermined period based on information on the opening of any one of the plurality of supply clamps 29 obtained from the culture device 11. This enables the sampling device 60 to store the same amount of sample from each of the plurality of reactors 12 in the temporary storage unit 136, making it possible to more reliably obtain a combined sample in which the individual samples are averaged.

[0091] The culture device 11 also includes a plurality of supply paths (EC supply circuits 44b) for supplying culture medium connected to each of the plurality of reactors 12, as well as a plurality of supply clamps 29 for opening and closing each of the plurality of supply paths, and the control unit (controller 68) operates the pumps (upstream pump 142, introduction pump 144) for one cycle period which is the sum of the open periods of all of the plurality of supply clamps 29. This allows the sampling device 60A to easily obtain a joint sample in which each sample is averaged, without obtaining information on the open status of the supply clamps 29 from the culture device 11.

[0092] The control unit (controller 68) also executes one cycle period for operating the pumps (upstream pump 142, introduction pump 144) two or more times. This allows the sampling device 60A to store a sufficient amount of combined sample in the temporary storage unit 136.

[0093] The pump is an upstream pump 142 provided in the sample introduction path 130 between the culture device 11 and the temporary storage section 136, and the control section (controller 68) operates the upstream pump 142 in a temporary storage step in which samples from each of the multiple reactors 12 flow into the temporary storage section 136, and stops the operation of the upstream pump 142 in a combined sample outflow step in which the combined sample flows out from the temporary storage section 136 to the sampling path 64. This allows the sampling device 60 to stably store each sample from each of the multiple reactors 12 in the temporary storage section 136 in the temporary storage step.

[0094] The sample introduction path 130 has a downstream clamp 146 between the sampling path 64 and the temporary storage section 136 that opens and closes the sample introduction path 130, and the control section (controller 68) closes the downstream clamp 146 in the temporary storage step and opens the downstream clamp 146 in the joint sample outflow step. This allows the sampling devices 60, 60A to prevent the sample from outflowing from the temporary storage section 136 in the temporary storage step.

[0095] The sample introduction path 130 also includes an introduction pump 144 between the sampling path 64 and the temporary storage section 136, and the control section (controller 68) stops the operation of the introduction pump 144 in the temporary storage step, and operates the introduction pump 144 in the combined sample discharge step to discharge the combined sample into the sampling path 64. This allows the sampling devices 60, 60A to smoothly introduce the combined sample into the sampling path 64 in the combined sample discharge step.

[0096] The pump is an introduction pump 144 provided in the sample introduction path 130 between the sampling path 64 and the temporary storage section 136, and the control section (controller 68) operates the introduction pump 144 in a temporary storage step in which samples from each of the plurality of reactors 12 flow into the temporary storage section 136, and also operates the introduction pump 144 in a combined sample outflow step in which the combined sample flows out from the temporary storage section 136 to the sampling path 64. As a result, the sampling device 60B can introduce the samples from each of the plurality of reactors 12 into the temporary storage section 136 by applying negative pressure to the sample introduction path 130 upstream of the introduction pump 144 while the introduction pump 144 is operating in the temporary storage step. Furthermore, the sampling device 60B achieves cost reduction by omitting the upstream pump 142.

[0097] Furthermore, the sample introduction path 130 has an upstream clamp 150 between the culture device 11 and the temporary storage section 136 that opens and closes the sample introduction path 130, and the control section (controller 68) opens the upstream clamp 150 in the temporary storage step and closes the upstream clamp 150 in the combined sample outflow step. This allows the sampling device 60B to store samples from each of the multiple reactors 12 in the temporary storage section 136 in the temporary storage step and to introduce the combined sample into the sampling path 64 in the combined sample outflow step.

[0098] In addition, a second aspect of the present invention is a cell culture system 10 that collects a liquid sample from a culture section (culture device 11) having a plurality of reactors 12 that culture cells based on the flow of culture medium, and the culture section supplies culture medium to the plurality of reactors 12 in sequence, and is equipped with a sampling path 64 through which the sample flows, a detection section 75 provided in the sampling path 64, a sample introduction path 130 that connects the sampling path 64 upstream of the detection section 75 and the culture section, pumps (upstream pump 142, introduction pump 144) that circulate the sample through the sample introduction path 130, and a control section (controller 68) that operates the pump, and has a temporary storage section 136 provided in the sample introduction path 130 that can temporarily store the sample, and the temporary storage section 136 combines the plurality of samples into a combined sample by sequentially flowing samples from each of the plurality of reactors 12 into it under the operation of the pump, and then discharges the combined sample into the sampling path 64. This allows the cell culture system 10 to detect samples from multiple reactors 12 more efficiently.

Claims

1. A sampling device for collecting a liquid sample from a culture device having a plurality of reactors for culturing cells based on the flow of a culture medium, a sampling path through which the sample flows; a detection unit provided in the sampling path; a sample introduction path that connects the sampling path upstream of the detection unit and the culture device; a pump for circulating the sample through the sample introduction path; a control unit that operates the pump, a temporary storage section provided in the sample introduction path and capable of temporarily storing the sample; The temporary storage unit sequentially receives the samples from the plurality of reactors under the operation of the pump, combines the plurality of samples into a combined sample, and discharges the combined sample into the sampling path. Sampling device.

2. 2. The sampling device according to claim 1, the culture device includes a plurality of supply paths for supplying culture medium connected to each of the plurality of reactors, and a plurality of supply clamps for opening and closing each of the plurality of supply paths; The control unit operates the pump for a predetermined period based on acquisition of open information of any one of the plurality of supply clamps from the culture device. Sampling device.

3. 2. The sampling device according to claim 1, the culture device includes a plurality of supply paths for supplying culture medium connected to each of the plurality of reactors, and a plurality of supply clamps for opening and closing each of the plurality of supply paths; The control unit operates the pump for one cycle period that is the sum of all open periods of the plurality of supply clamps. Sampling device.

4. 4. The sampling device according to claim 3, The control unit executes the one cycle period for operating the pump two or more times. Sampling device.

5. The sampling device according to any one of claims 1 to 4, the pump is an upstream pump provided in the sample introduction path between the culture device and the temporary reservoir, The control unit operates the upstream pump in a temporary storage step of causing the samples from each of the plurality of reactors to flow into the temporary storage unit, and stops operation of the upstream pump in a combined sample outflow step of causing the combined sample to flow out from the temporary storage unit to the sampling path. Sampling device.

6. 6. The sampling device according to claim 5, the sample introduction path has a downstream clamp between the sampling path and the temporary reservoir that opens and closes the sample introduction path; The control unit closes the downstream clamp in the temporary storage step and opens the downstream clamp in the combined sample outflow step. Sampling device.

7. 7. The sampling device according to claim 5 or 6, the sample introduction path includes an introduction pump between the sampling path and the temporary storage section; The control unit stops the operation of the introduction pump in the temporary storage step, and operates the introduction pump in the combined sample outflow step to outflow the combined sample into the sampling path. Sampling device.

8. The sampling device according to any one of claims 1 to 4, the pump is an introduction pump provided in the sample introduction path between the sampling path and the temporary storage section, The control unit operates the introduction pump in a temporary storage step of causing the samples for each of the plurality of reactors to flow into the temporary storage unit, and operates the introduction pump in a combined sample outflow step of causing the combined sample to flow out from the temporary storage unit to the sampling path. Sampling device.

9. 9. The sampling device according to claim 8, the sample introduction path has an upstream clamp between the culture device and the temporary reservoir that opens and closes the sample introduction path; The control unit opens the upstream clamp in the temporary storage step and closes the upstream clamp in the combined sample outflow step. Sampling device.

10. A cell culture system for collecting a liquid sample from a culture unit having a plurality of reactors for culturing cells based on the flow of a culture medium, The culture unit sequentially supplies culture media to the plurality of reactors, a sampling path through which the sample flows; a detection unit provided in the sampling path; a sample introduction path connecting the sampling path upstream of the detection unit and the culture unit; a pump that circulates the sample through the sample introduction path; a control unit that operates the pump, a temporary storage section provided in the sample introduction path and capable of temporarily storing the sample; The temporary storage unit sequentially receives the samples from the plurality of reactors under the operation of the pump, combines the plurality of samples into a combined sample, and discharges the combined sample into the sampling path. Cell culture system.

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