Sampling method

Abstract

This sampling device (60) comprises a sampling route (64), a detection unit (75), and a sample introduction route (130). A culture device (11) or the sample introduction route (130) comprises a sterile filter (136) in a segment ending at a location at which a sample is introduced into the sampling route (64). A sampling method according to the present invention includes a sampling step in which a sample from the culture device (11) is introduced and detection is performed by the detection unit (76), and an attached matter removal step in which attached matter attached to the sterile filter (136) as a result of a fluid being made to flow through the sample introduction route (130) from the sampling route (64) is removed.

Description

Technical Field

[0001] The present invention relates to a sampling method for collecting a liquid sample of a cell culture apparatus.

Background Art

[0002] For example, U.S. Patent No. 9,442,047 discloses a sampling device having a sampling path for collecting a liquid sample from a culture device. The sampling device includes a pump that draws a sample (culture medium) 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 contained components and component amounts (concentrations) of the sample.

Summary of the Invention

[0003] By the way, when applying this type of sampling device, in order to prevent bacteria from entering the culture device from the sampling device, a sterile filter is installed in the sample outflow path of the culture device. However, since the culture medium is periodically collected during cell culture, the sterile filter gradually clogs due to the adhesion of aggregates such as proteins contained in the culture medium. Thus, when the sterile filter becomes clogged, inconveniences such as the inability to collect a specified amount of sample, the negative pressure in the sample introduction path, and the change in gas concentration (generation of bubbles) occur.

[0004] In view of the above circumstances, an object of the present invention is to provide a sampling method capable of satisfactorily collecting a sample by improving the clogging of deposits even in a configuration provided with a sterile filter.

[0005] To achieve the above objective, one aspect of the present invention is a sampling method for collecting a liquid sample from a culture section for culturing cells to a sampling section, wherein the sampling section comprises a sampling path through which the sample flows, a detection unit provided in the sampling path so as to be in contact with the sample, and a sample introduction path connecting the culture section and the sampling path upstream of the detection unit, the culture section or the sample introduction path is equipped with a sterile filter in the section up to the introduction of the sample into the sampling path, and the method comprises a sampling step of introducing the sample from the culture section to the sampling path via the sample introduction path and detecting the sample with the detection unit, and a deposit removal step of removing deposits attached to the sterile filter by flowing a fluid from the sampling path to the sample introduction path.

[0006] The sampling method described above can improve filter clogging, thereby enabling better sample collection. [Brief explanation of the drawing]

[0007] [Figure 1] This is a schematic perspective view 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] This is a schematic diagram illustrating the pathway of the culture medium during cell culture. [Figure 3] This is an explanatory diagram illustrating the sampling device's path in general terms. [Figure 4] This is a flowchart showing the sampling method of the sampling device. [Figure 5] This is an explanatory diagram showing the operation of the priming and washing processes. [Figure 6] This is a flowchart of the sampling process. [Figure 7] This is an explanatory diagram showing the operation of the main process. [Figure 8]Figure 8A is an explanatory diagram showing the behavior of aggregates on the sterile filter in the first stage. Figure 8B is an explanatory diagram showing the behavior of aggregates on the sterile filter in the second stage. [Figure 9] This is an explanatory diagram showing the operation of the deposit removal process. [Figure 10] This is an explanatory diagram illustrating the path of the sampling device according to the second embodiment. [Figure 11] Figure 10 is an explanatory diagram showing the cleaning process and the process for removing attached substances from the sampling device. [Figure 12] This is an explanatory diagram illustrating the path of the sampling device according to the third embodiment. [Figure 13] Figure 12 is an explanatory diagram showing the cleaning process of the sampling device. [Figure 14] Figure 12 is an explanatory diagram showing the process of removing deposits from the sampling device. [Figure 15] This is an explanatory diagram illustrating the path of the sampling device according to the fourth embodiment. [Figure 16] Figure 15 is an explanatory diagram showing the main process of the sampling apparatus. [Figure 17] Figure 15 is an explanatory diagram showing the process of removing deposits from the sampling device. [Modes for carrying out the invention]

[0008] The present invention will be described in detail below with reference to the attached drawings, with reference to preferred embodiments.

[0009] [First Embodiment] As shown in Figure 1, the sampling device 60 according to the first embodiment of the present invention is applied to a cell culture system 10 used to culture living cells in regenerative medicine. The sampling device 60 samples the culture medium during cell culture by the cell culture system 10 and measures the state of the culture medium. For example, the cell culture system 10 continues cell culture over a long period of time by supplying culture medium and oxygen to the reactor 12, which is a cell culture vessel, and by discharging lactic acid, carbon dioxide, etc. (including unused culture medium and oxygen) generated during cell culture from the reactor 12.

[0010] The cells of a living organism are not particularly limited, but examples include cells contained in blood (such as T cells) and stem cells (such as ES cells, iPS cells, and mesenchymal stem cells). The culture medium should also be selected appropriately according to the type of living organism's cells. For example, a balanced salt solution (BSS) can be used as the basic solution, to which various amino acids, vitamins, and serum can be added.

[0011] The cell culture system 10 includes a culture device 11 (culture section) in which a reactor 12 is set and cells are actually cultured, and a sampling device 60 (sampling section) for taking a liquid sample from the culture device 11 during culture. In Figure 1, a culture device 11 equipped with one reactor 12 is shown, but the number of reactors 12 is not particularly limited, and the culture device 11 may be equipped with multiple reactors 12. The cell culture system 10 may also be configured in which multiple culture devices 11 are connected to one sampling device 60. In this embodiment, a cell culture system 10 in which the culture section and the sampling section are configured as separate units is illustrated, but the cell culture system 10 may be a device in which the culture section and the sampling section are integrated (combined).

[0012] The culture apparatus 11 includes a culture medium storage section 14 for storing culture medium, a flow path 16 provided between the reactor 12 and the culture medium storage section 14, a plurality of medical bags 18 connected to the flow path 16, and a waste liquid section 20 for storing liquid discharged from the flow path 16.

[0013] A hard tank capable of storing a large amount of culture medium is applied to the culture medium storage unit 14. The flow path 16 is composed of a plurality of tubes 22, and each tube 22 is connected to each of the reactor 12, the culture medium storage unit 14, the plurality of medical bags 18, and the waste liquid unit 20.

[0014] Examples of the plurality of medical bags 18 include a cell fluid bag 18A storing a liquid containing cells (cell fluid), a washing liquid bag 18B storing a washing liquid, a detachment liquid bag 18C storing a detachment liquid, and a recovery bag (not shown) for recovering cultured cells. The washing liquid is a liquid used during priming of the reactor 12 and the flow path 16. Examples of this washing liquid include buffer solutions such as PBS (Phosphate Buffered Salts) and TBS (Tris - Buffered Saline), or physiological saline. The detachment liquid is a liquid for detaching cells cultured by a culture process. As the detachment liquid, for example, trypsin or an EDTA solution can be applied.

[0015] When constructing the cell culture system 10, the flow path 16 is set to pass through the flow path control mechanism unit 24 of the culture device 11. The flow path control mechanism unit 24 has a housing 26 that houses a part of the flow path 16. Also, the flow path control mechanism unit 24 includes, in the housing 26, a clamp 28 for opening and closing a predetermined tube 22, a pump 30 for circulating the liquid in the tube 22, and a control circuit 32 for controlling the operations of the clamp 28 and the pump 30 (see FIG. 2).

[0016] The reactor 12 is housed within the housing 26 of the flow path control mechanism section 24. The reactor 12 includes a plurality (for example, 10,000 or more) of hollow fibers 34 and a case 36 that houses the plurality of hollow fibers 34. Each hollow fiber 34 has a lumen (not shown), and cells are seeded on the inner peripheral 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, and each pore allows a solution or a low-molecular-weight substance to permeate therethrough without allowing cells or proteins to permeate. A medium or the like is supplied to the cells seeded on the inner peripheral surface of the hollow fiber 34 through the lumen or the pores. Hereinafter, a configuration in which a liquid flows mainly through the lumen of the hollow fiber 34 is also referred to as IC (intra capillary), and a configuration in which a liquid flows mainly outside the hollow fiber 34 is also referred to as EC (extra capillary).

[0017] The case 36 includes a first IC terminal 36a and a second IC terminal 36b that communicate with the lumen of the hollow fiber 34, and a first EC terminal 36c and a second EC terminal 36d that communicate with the space outside the hollow fiber 34 within the case 36, and a tube 22 is connected to each terminal.

[0018] Hereinafter, referring to FIG. 2, the flow path 16 between the reactor 12 and the medium reservoir 14 and the configuration of the flow path control mechanism section 24 will be specifically described. The flow path 16 includes a medium delivery route 40 connected to the medium reservoir 14, and an IC route 42 (internal route) and an EC route 44 (external route) branched from the medium delivery route 40. The IC route 42 is a route for supplying a liquid to the lumen of the hollow fiber 34. The EC route 44 is a route for supplying a liquid into the case 36 outside the hollow fiber 34.

[0019] The IC route 42 includes an IC circulation circuit 42a that can circulate liquid to and from the reactor 12, and an IC supply circuit 42b that can circulate 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 in the lumen of the hollow fiber 34. Downstream of the reactor 12 in the IC circulation circuit 42a, an IC waste liquid circuit 46 is connected that discharges the culture medium to the waste liquid section 20. On the other hand, 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.

[0020] On the other hand, the EC route 44 has an EC circulation circuit 44a that can circulate liquid to and from the reactor 12, and an EC supply circuit 44b that can circulate 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 is equipped with an EC circulation pump 30c that circulates liquid outside the hollow fiber 34. Upstream of the reactor 12 in the EC circulation circuit 44a, a gas exchanger 52 is provided. The gas exchanger 52 discharges carbon dioxide mixed in with the culture medium while mixing a predetermined gas component (for example, nitrogen N2: 75%, oxygen O2: 20%, carbon dioxide CO2: 5%) into the culture medium. Downstream of the reactor 12 in the EC circulation circuit 44a, an EC waste liquid circuit 48 is connected that discharges the culture medium to the waste liquid section 20. The EC supply circuit 44b is equipped with an EC supply pump 30d that circulates liquid from the culture medium delivery route 40 to the EC circulation circuit 44a.

[0021] Although not shown in the diagram, in addition to the culture medium storage section 14, multiple medical bags 18 (cell solution bag 18A, washing solution bag 18B, and detachment solution bag 18C) are connected via multiple tubes 22 to the IC supply circuit 42b upstream of the IC supply pump 30b, or to the EC supply circuit 44b upstream of the EC supply pump 30d. These medical bags 18 may be replaced with recovery bags, etc., using a sterile joining device that sterilizes and joins the bags, depending on the application.

[0022] The sampling device 60 is connected to the EC circulation circuit 44a of the culture device 11 at a location near the downstream side (second EC terminal 36d) of the reactor 12 (between the reactor 12 and the EC waste liquid circuit 48). Therefore, one end of the sample outflow path 54, which discharges the culture medium, a liquid sample, is connected to the EC circulation circuit 44a. The other end of the sample outflow path 54 is provided with a culture device side connector 56. 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 sample outflow path 54 may also be connected to the downstream side (second IC terminal 36b) of the reactor 12 in the IC circulation circuit 42a.

[0023] Next, the configuration of the sampling device 60 will be described with reference to Figure 3. The sampling device 60 collects a sample of culture medium from one or more culture devices 11 and detects the components and their amounts (concentrations) in the sample. The sampling device 60 comprises a sampling kit 62 having a sampling path 64 through which the sample is collected, a plurality of mechanical parts 66 to which the sampling kit 62 is detachably set, and a controller 68 that controls the operation of the plurality of mechanical parts 66. The sampling kit 62 is a disposable item, and the plurality of mechanical parts 66 are reusable items.

[0024] The sampling kit 62 includes, in addition to the sampling path 64, a washing liquid container 70, a standard liquid container 72, a waste liquid container 74, and a detection unit 75 (first detection unit 76, second detection unit 80). The sampling path 64 is made of a flexible tube of an appropriate diameter that can allow the sample to flow through. The washing liquid container 70 is connected to a branching point 65 to which one end of the sampling path 64 is connected via a washing liquid branching passage 71, and the standard liquid container 72 is connected to this branching point 65 via a standard liquid branching passage 73. The waste liquid container 74 is connected to the other end of the sampling path 64.

[0025] The cleaning solution storage section 70 and the standard solution storage section 72 are formed in the shape of a bag (medical bag) from a soft resin material such as polyvinyl chloride or polyolefin. However, the cleaning solution storage section 70 and the standard solution storage section 72 are not particularly limited as long as they are capable of holding liquid. The waste liquid storage section 74 shares the tank of the waste liquid section 20 of the culture apparatus 11, but is not limited to this, and medical bags or the like may be used.

[0026] The washing solution container 70 contains the washing solution. The washing solution is not particularly limited, and for example, buffer solutions, physiological saline, etc., as listed as the washing solution for the washing solution bag 18B of the culture device 11 may be used as appropriate.

[0027] The standard solution container 72 contains a standard solution. The standard solution is a liquid used to calibrate the first detection unit 76 and the second detection unit 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.

[0028] The first detection unit 76 and the second detection unit 80 are provided in series and spaced apart from each other at an intermediate position in the sampling path 64. The detection unit 75 is not limited to a structure divided into the first detection unit 76 and the second detection unit 80; it may also be a structure in which the first detection unit 76 and the second detection unit 80 are integrated, or a structure divided into three or more units.

[0029] The first detection unit 76 is a cylindrical member having a plurality of first element units 78 that come into contact with (wet the liquid with) the sample in a flow path within the sampling path 64. For example, the plurality of first element units 78 include a pH chip 78a for measuring the pH in the sample, an O2 chip 78b for measuring the O2 concentration in the sample, and a CO2 chip 78c for measuring the CO2 concentration in the sample. The pH chip 78a is H + , OH - It reacts to the element and produces a color. The O2 tip 78b reacts to O2 and produces a color. The CO2 tip 78c reacts to CO2 and produces a color.

[0030] The second detection unit 80 is a cylindrical member having a plurality of second element units 82 that come into contact with (wet the liquid with) the sample in a flow path within the sampling path 64, and is located downstream of the first detection unit 76 (towards the waste liquid containment unit 74). For example, the plurality of second element units 82 are biosensors that react enzymes with the flowing sample to detect changes in current, etc. Examples of the plurality of second element units 82 include glucose chips 82a for measuring glucose concentration in the sample and lactate chips 82b for measuring lactate concentration in the sample. The glucose chip 82a is electrically connected to a glucose terminal 83a that protrudes to the outside of the cylindrical member. The lactate chip 82b is electrically connected to a lactate terminal 83b that protrudes to the outside of the cylindrical member.

[0031] Furthermore, the sampling kit 62 includes a connection portion 84 between the branching 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 portion 84 is, for example, a component integrally molded from multiple branching ports, each equipped with a valve (not shown) that closes when the sample introduction path 130 is not installed and opens when the sample introduction path 130 is installed (in Figure 3, the connection portion 84 is shown for convenience as an area enclosed by a dashed line). Alternatively, the connection portion 84 can be configured to include ports to which the sample introduction path 130 can be connected while ensuring the sterility of the sampling path 64.

[0032] As shown in Figure 3, a portion of the sampling kit 62 is set in the main mechanism 90, which is one of the multiple mechanism units 66. The main mechanism 90 is housed in a housing 91 (see Figure 1) and includes a main mechanism pump 92 and multiple clamps 94 that open and close the flow paths in each path (tube). Although not shown, a controller 68 for controlling the sampling device 60 may also be provided in the main mechanism 90. When the sampling kit 62 is set in the main mechanism 90, the main unit 96 of the sampling device 60 is constructed.

[0033] The pump 92 on the main mechanism side is equipped with a sampling path 64 that extends between the branching point 65 and the connection point 84. The pump 92 on the main mechanism side has a circular winding part that can wrap around the sampling path 64, and rotates in a way that squeezes the winding sampling path 64 (tube) to circulate the fluid (liquid, air, etc.) inside.

[0034] The multiple clamps 94 include a cleaning liquid clamp 94a that opens and closes the cleaning liquid branching passage 71, a standard liquid clamp 94b that opens and closes the standard liquid branching passage 73, and a waste liquid clamp 94c that opens and closes the sampling path 64 between the second detection unit 80 and the waste liquid storage unit 74.

[0035] Furthermore, the first detection unit 76 of the sampling kit 62 is set in the first measuring instrument 110, which is one of the multiple mechanical parts 66, thereby constructing the first sensor unit 111. The first measuring instrument 110 has a holder 112 that houses the first detection unit 76, and a measuring body 116 fixed to the holder 112 that optically measures the multiple first element parts 78.

[0036] The measuring main unit 116 has a PH detector 116a, an O2 detector 116b, and a CO2 detector 116c, facing the PH chip 78a, O2 chip 78b, and CO2 chip 78c, while the first detection unit 76 is held in the holder 112. Under the control of the controller 68, the measuring main unit 116 emits measurement light of a wavelength corresponding to the characteristics of each first element unit 78, and receives the excitation light generated from the excitation of each first element unit 78, thereby transmitting the detection signal to the controller 68. The measuring main unit 116 is calibrated by the user when it is set in a calibration device 118 (see Figure 1) installed adjacent to the main mechanism unit 90.

[0037] Furthermore, the second detection unit 80 of the sampling kit 62 is set in the second measuring instrument 120, which is one of the multiple mechanical parts 66, thereby constructing the second sensor unit 121. The second measuring instrument 120 has a case 122 capable of housing the second detection unit 80 and an enzyme detector (not shown) electrically connected to the glucose terminal 83a and the lactate terminal 83b. The enzyme detector detects current values ​​from the glucose chip 82a and the lactate chip 82b, respectively, and transmits a detection signal based on the current value to the controller 68.

[0038] Then, in order to introduce the sample to be measured by the first sensor unit 111 and the second sensor unit 121, a sample introduction path 130 is connected to the connection part 84 of the sampling kit 62 (sampling path 64). The sample introduction path 130, like the sampling path 64, is made up of a flexible tube of an appropriate diameter that can allow the sample to flow through.

[0039] The sample introduction path 130 has a sampling device side connector 132 at one end for connecting to the culture device side connector 56 (see also Figure 2). The other end of the sample introduction path 130 is provided with a detachable plug (not shown) that can be attached to a connection part 84. Hereinafter, the point where the plug of the sample introduction path 130 is connected to the sampling path 64 will be referred to as the connection point 134. The sampling path 64 and the sample introduction path 130 may be inseparably connected to each other at the connection part 84.

[0040] Furthermore, a sterile filter 136 is provided in the sample introduction path 130 between the sampling device connector 132 and the plug (connection point 134). The sterile filter 136 maintains the sterile state of the sample introduction path 130 and the culture device 11 upstream of the sterile filter 136.

[0041] For example, the sterile filter 136 has a housing 136a connected to the sample introduction path 130, and a mesh body 136b housed in a space within the housing that communicates with the flow path of the sample introduction path 130 (see Figure 8A). The mesh body 136b may be a membrane filter or depth filter capable of capturing bacteria of an appropriate size.

[0042] Furthermore, a waste liquid route 138 is connected to the sample introduction route 130 upstream of the sterile filter 136. The waste liquid route 138 connects a branching point 131 of the sample introduction route 130, which is located between the sterile filter 136 and the sampling device side connector 132, to a branching point 86 of the sampling route 64 downstream of the second detection unit 80 (waste liquid clamp 94c). The waste liquid route 138 circulates the washing liquid introduced into the sample introduction route 130 to the waste liquid storage unit 74.

[0043] The wastewater route 138 is equipped with a sterile filter 139 near the branching point 131 (closer to the sample introduction route 130). The sterile filter 139 uses the same filter as the sterile filter 136 to maintain the sterile state of the culture device 11.

[0044] A portion of the sample introduction path 130 is detachably set in an introduction mechanism 140, which is one of a plurality of mechanism units 66. The introduction mechanism 140 includes an introduction pump 142 and a waste liquid path clamp 144. Furthermore, the introduction mechanism 140 has a pressure sensor 146 for detecting the pressure in the flow path of the sample introduction path 130, and a bubble sensor 148 for detecting bubbles in the flow path of the sample introduction path 130. When the sample introduction path 130 is set in the introduction mechanism 140, the introduction unit 141 of the sampling device 60 is constructed.

[0045] The introduction unit 141 allows for the integrated handling of a portion of the sample introduction path 130, the introduction pump 142, the pressure sensor 146, and the bubble sensor 148. A short portion of the sample introduction path 130 extending from the introduction unit 141 is connected to the connection point 84 on the main unit 96.

[0046] The introduction pump 142 is positioned upstream of the branching point 131 in the sample introduction path 130 (between the branching point 131 and the sampling device side connector 132). The introduction pump 142 has a circular wrapping portion that can wrap around the sample introduction path 130 so that it wraps around it, and rotates to squeeze the wrapping sample introduction path 130 (tube), thereby circulating the fluid inside.

[0047] The wastewater path clamp 144 is positioned in the wastewater path 138 between the sterile filter 139 and the branching point 86 of the sampling path 64. The wastewater path clamp 144 switches the outflow and stopflow of the cleaning fluid through the wastewater path 138 by opening and closing the wastewater path 138.

[0048] The pressure sensor 146 is positioned in the sample introduction path 130 between the branching point 131 and the sterile filter 136 (upstream of the sterile filter 136), and detects the internal pressure of the sample introduction path 130 at this location. The detection result detected by the pressure sensor 146 is wirelessly transmitted to the controller 68. In order to improve the pressure detection accuracy of the pressure sensor 146, the planned location of the pressure sensor 146 in the sample introduction path 130 may be formed into an appropriate shape (such as a cylindrical or disc shape with a larger diameter than other locations).

[0049] Similarly, the bubble sensor 148 is positioned in the sample introduction path 130 between the connection point 134 and the pressure sensor 146 (upstream of the sterile filter 136) to detect bubbles in the sample introduction path 130. The detection result from the bubble sensor 148 is wirelessly transmitted to the controller 68. The bubble sensor 148 may also be positioned upstream of the pressure sensor 146 (towards the branching point 131).

[0050] The controller 68 (control unit) is a computer having one or more processors, memory, input / output interfaces, and electronic circuits (not shown). The controller 68 controls the entire sampling device 60 by having the processor execute a program stored in memory. In this embodiment, the controller 68 is configured to communicate with the control circuit 32 of the culture device 11, and performs linked control of the culture device 11 and the sampling device 60. The controller 68 may also be a control device integrated with the control circuit 32 of the culture device 11.

[0051] 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 Figure 4. The sampling method consists of sequentially performing a preparation step, a priming step, a sampling step, a cleaning step, and a calibration step.

[0052] First, in the preparation step (step S1), the user of the cell culture system 10 sets (attaches) the sampling kit 62 to the main mechanism 90 to form the main unit 96, as shown in Figure 3. The user also sets the first detection unit 76, which is exposed from the housing 91, to the first measuring instrument 110 to construct the first sensor unit 111, and sets the second detection unit 80, which is also exposed, to the second measuring instrument 120 to construct the second sensor unit 121. These first sensor unit 111 and second sensor unit 121 are suspended from the stand 98.

[0053] Furthermore, the user sets the sample introduction path 130 into the introduction mechanism 140 to form the introduction unit 141. Subsequently, the user connects the sampling device side connector 132 of the sample introduction path 130, which is exposed from the introduction unit 141, to the culture device side connector 56, and also connects the plug of the sample introduction path 130 to the connection part 84.

[0054] Next, in the priming process (step S2 in Figure 4), the controller 68 opens the cleaning solution clamp 94a and the waste liquid clamp 94c, as shown in Figure 5, while closing the standard solution clamp 94b and the waste liquid path clamp 144. In this state, the controller 68 rotates the main mechanism pump 92. This creates negative pressure in the cleaning solution branch 71, and cleaning solution is supplied from the cleaning solution container 70. The cleaning solution that has passed through the cleaning solution branch 71 and the branching point 65 passes through the main mechanism pump 92 in the sampling path 64, flows sequentially through the connection point 84, the first detection unit 76, and the second detection unit 80, and is discharged into the waste liquid container 74. Also, in the priming process, the introduction pump 142 is stopped, so that cleaning solution does not flow into the sample introduction path 130.

[0055] Next, in the sampling process (step S3 in Figure 4), the sampling device 60 guides the sample from the culture device 11 to the sampling path 64, and the detection unit 75 detects the components and amounts of the components in the sample. At this time, as shown in Figure 6, the controller 68 first performs the main process (step S3-1). In this main process, as shown in Figure 7, the controller 68 closes the washing solution clamp 94a, the standard solution clamp 94b, and the waste liquid path clamp 144, while opening the waste liquid clamp 94c. The controller 68 also stops the rotation of the main mechanism pump 92 while rotating the introduction pump 142. As a result, negative pressure is applied to the sample introduction path 130 upstream of the introduction pump 142, and the sample is introduced from the culture device 11.

[0056] The sample drawn in from the culture device 11 passes through the sterile filter 136 as it flows through the sample introduction path 130. At this time, aggregates of proteins and other substances contained in the sample are captured by the sterile filter 136. To enhance the rinsing effect of the sterile filter 136 on these aggregates, the controller 68 controls the rotation speed of the introduction pump 142 when the main process is being executed.

[0057] Specifically, in the first stage after the start of the main process, the controller 68 rotates the introduction pump 142 at a fast first rotational speed to flow the sample at a first flow rate. In the second stage after the first stage, the controller 68 rotates the introduction pump 142 at a second rotational speed slower than the first rotational speed to flow the sample at a second flow rate slower than the first flow rate.

[0058] The ratio of the first flow velocity (first rotation speed) to the second flow velocity (second rotation speed) is preferably set to, for example, 3 to 10 times. The actual value of the first flow velocity is preferably set within the range of approximately 20 mL / min to 60 mL / min, and the actual value of the second flow velocity is preferably set within the range of approximately 1 mL / min to 10 mL / min. As a result, as shown in Figure 8A, in the first stage, aggregates on the filter surface on the sampling path 64 side that were captured in the previous deposit removal step are allowed to flow into the sampling path 64. Then, in the second stage, as shown in Figure 8B, by slowly flowing the sample and guiding it to the sampling path 64, it is possible to reduce the excessive capture of aggregates on the filter surface on the culture device 11 side.

[0059] Furthermore, it is preferable that the controller 68 sets the execution period of the first stage to a few seconds (for example, 1 to 3 seconds), and the execution period of the second stage to be set so that a predetermined amount of sample can be obtained. By performing the first stage in a short period after the start of the main process, it is possible to enhance the effect of washing away aggregates captured on the filter surface on the sampling path 64 side, while reducing the excessive capture of aggregates on the filter surface on the culture device 11 side. Note that the controller 68 may be configured to change the sample flow rate (rotation speed of the introduction pump 142) in steps of 3 or more, or it may be configured to change it linearly from the first flow rate to the second flow rate.

[0060] Returning to Figure 7, the sample in the sample introduction path 130, having passed through the sterile filter 136, flows sequentially through the connection section 84 (connection point 134), the first detection unit 76, and the second detection unit 80 before being discharged into the waste liquid collection unit 74. As the sample passes through, the multiple first element units 78 (PH tip 78a, O2 tip 78b, CO2 tip 78c) of the first detection unit 76 come into contact with the sample and change color according to the respective contents of PH, O2, and CO2. The first measuring instrument 110 performs optical measurements on each first element unit 78 and transmits the detection results to the controller 68. Upon receiving the detection results, the controller 68 performs appropriate processing and displays the measured values ​​(PH value, O2 concentration, CO2 concentration) on the monitor 100 of the main mechanism unit 90.

[0061] Similarly, as the sample passes through, the multiple second element units 82 (glucose chip 82a, lactic acid chip 82b) of the second detection unit 80 come into contact with the sample, and the second measuring instrument 120 detects current values ​​corresponding to the glucose and lactic acid content. The second measuring instrument 120 transmits each detection result to the controller 68. Upon receiving the detection results, the controller 68 performs appropriate processing and displays the measured values ​​(glucose concentration, lactic acid concentration) on the monitor 100.

[0062] Returning to Figure 6, the controller 68 determines whether or not to terminate the main process while it is being performed (step S3-2). For example, the controller 68 determines the end of the sampling process based on whether the detection unit 75 has finished detecting the sample and displaying the detection result, or whether the sampling process has been performed for a predetermined time.

[0063] Furthermore, during the execution of the main process, the controller 68 detects the pressure in the sample introduction path 130 using the pressure sensor 146 and detects air bubbles in the sample introduction path 130 using the air bubble sensor 148 (step S3-3). Based on the detected pressure and air bubbles, the controller 68 determines whether the sterile filter 136 is clogged by the sample flowing through the sample introduction path 130 (step S3-4). For example, the controller 68 determines that the sterile filter 136 is clogged when the pressure detected by the pressure sensor 146 falls below a predetermined pressure threshold (not shown). Alternatively, the controller 68 determines that the sterile filter 136 is clogged when the air bubble sensor 148 detects that the sample contains more than a predetermined amount of air bubbles during the sampling process.

[0064] If the sterile filter 136 is not clogged (Step S3-4: NO), the process returns to Step S3-1 and the same processing flow is repeated. If the sterile filter 136 is found to be clogged (Step S3-4: YES), the controller 68 stops the main process and performs the deposit removal process (Step S3-5) to remove aggregates from the sterile filter 136. As shown in Figure 9, in the deposit removal process, the controller 68 closes the standard solution clamp 94b and the waste liquid clamp 94c, while opening the washing solution clamp 94a and the waste liquid path clamp 144. The controller 68 also stops the rotation of the introduction pump 142 while rotating the main mechanism pump 92.

[0065] Furthermore, during the deposit removal process, the controller 68 maintains a constant rotational speed of the main mechanism pump 92, introducing the cleaning solution into the sample introduction path 130 at a constant flow rate. As a result, the cleaning solution in the cleaning solution container 70 is introduced into the sampling path 64 via the cleaning solution branch 71. Because the waste liquid clamp 94c is closed, this cleaning solution does not go towards the detection unit 75, but flows into the sample introduction path 130 from the connection point 134. The cleaning solution that flows into the sample introduction path 130 passes from the sampling path 64 side to the culture device 11 side in the sterile filter 136, thereby removing aggregates attached to the sterile filter 136.

[0066] Then, with the wastewater path clamp 144 open and the introduction pump 142 stopped, the cleaning liquid containing the aggregate flows from the branching point 131 to the wastewater path 138. In the wastewater path 138, the aggregate is removed as the cleaning liquid passes through the sterile filter 139. The cleaning liquid then returns from the wastewater path 138 to the sampling path 64 via the branching point 86, flows through the sampling path 64, and is discharged into the wastewater storage section 74.

[0067] In other words, the sampling device 60 can effectively clear clogging of the sterile filter 136 by performing a deposit removal process according to the state of the sample during the sampling process. After performing the deposit removal process for a predetermined period, the controller 68 returns to the main process (step S3-1) in Figure 6 and performs the main process from the beginning. As a result, the sampling device 60 can introduce a specified amount of sample into the sampling path 64 that can be properly detected by the detection unit 75.

[0068] Furthermore, the deposit removal process is not limited to being performed according to the condition of the sample during the sampling process; for example, it may be performed periodically after the sampling process, after the cleaning process described later, etc. Also, the controller 68 may, when performing the deposit removal process, control the rotation speed of the main mechanism pump 92 in the same way as the main process.

[0069] For example, in the first stage after the start of the deposit removal process, the controller 68 rotates the main mechanism pump 92 at a fast first rotational speed, thereby circulating the cleaning fluid at a first flow rate. In the second stage after the first stage, the controller 68 rotates the main mechanism pump 92 at a second rotational speed, which is slower than the first rotational speed, thereby circulating the cleaning fluid at a second flow rate, which is slower than the first flow rate. Furthermore, the duration of the first stage is shorter than the duration of the second stage. As a result, in the first stage, aggregates adhering to the sterile filter 136 are removed more easily, and aggregates captured in the previous deposit removal process are made more likely to pass through the sterile filter 139.

[0070] Returning to Figure 4, after the sampling process, the controller 68 determines whether or not the cell culture in the culture device 11 is complete (step S4). If the cell culture is not complete (step S4: NO), the washing process (step S5) is performed. In the washing process, the controller 68 supplies the washing solution from the washing solution container 70 to the sampling path 64, similar to the priming process shown in Figure 5. This removes the samples adhering to the multiple first element units 78 (PH tip 78a, O2 tip 78b, CO2 tip 78c) and the multiple second element units 82 (glucose tip 82a, lactic acid tip 82b) using the washing solution.

[0071] Furthermore, the sampling device 60 performs a calibration process (step S6) as needed. In the calibration process, the controller 68 opens the clamp 94b for the standard solution and the clamp 94c for the waste liquid, and closes the clamp 94a for the washing solution and the clamp 144 for the waste liquid path, while rotating the pump 92 on the main mechanism side. As a result, the standard solution in the standard solution storage section 72 is guided from the standard solution branching path 73 to the sampling path 64, and flows sequentially through the connection section 84, the first detection section 76, and the second detection section 80 before being discharged to the waste liquid storage section 74.

[0072] At this time, the second sensor unit 121 measures the glucose concentration and lactate concentration in the standard solution and transmits the measurement results to the controller 68 or the second measuring instrument 120. The controller 68 or the second measuring instrument 120 calibrates the second measuring instrument 120 based on the measurement results of the second sensor unit 121. Meanwhile, the first sensor unit 111 (first measuring instrument 110) is set in the calibration device 118 by the user. The first measuring instrument 110 then measures the standard solution, pH, O2 concentration, and CO2 concentration in the calibration device 118 and transmits the measurement results to the controller 68 or the first measuring instrument 110. The controller 68 or the first measuring instrument 110 calibrates the pH detector 116a, O2 detector 116b, and CO2 detector 116c based on these measurement results.

[0073] Once the washing process (or calibration process) is complete, the controller 68 returns to step S3 and sequentially performs the subsequent steps. Meanwhile, in step S4, if the controller 68 determines that the cell culture is complete (step S4: YES), it terminates the operation flow of the sampling device 60.

[0074] The sampling device 60 and sampling method are not limited to those described above, and various methods can be employed. For example, the waste liquid path 138 of the sampling device 60 may not be connected to the sampling path 64, but to a waste liquid recovery unit (not shown) different from the waste liquid containment unit 74. Also, the waste liquid path 138 may not be configured to include a sterile filter 139.

[0075] [Second Embodiment] The sampling device 60A according to the second embodiment differs from the sampling device 60 described above in that, as shown in Figure 10, the sample introduction path 130 does not have a waste liquid path 138, and the washing liquid is directed to the culture device 11. In the following description, elements having the same configuration or function as those in the above embodiment are denoted by the same reference numerals, and their detailed description is omitted.

[0076] In detail, the introduction unit 141 of the sampling device 60A has an introduction pump 142 positioned near the connection point 134 of the sample introduction path 130. The introduction pump 142 is rotatable in a first direction to guide the sample to the sampling path 64 and in a second direction to draw the washing liquid into the sample introduction path 130.

[0077] Furthermore, the introduction unit 141 has a pressure sensor 146 and a bubble sensor 148 positioned in the sample introduction path 130 upstream of the introduction pump 142. The culture device 11 is equipped with a sterile filter 58 to maintain a sterile state on the culture device 11 side. The sterile filter 58 is installed between the EC circulation circuit 44a and the culture device side connector 56 (see also Figure 2).

[0078] The sampling device 60A according to the second embodiment is basically configured as described above, and its operation will be explained below. In the sampling method using the sampling device 60A, each step in the processing flow of Figure 4, except for the washing step, basically performs the same operation (however, since there is no waste liquid path clamp 144, this clamp is not opened or closed).

[0079] For example, in the sampling process (main process), the controller 68 of the sampling device 60A closes the washing solution clamp 94a and the standard solution clamp 94b, while opening the waste liquid clamp 94c. The controller 68 also stops the rotation of the main mechanism pump 92, while rotating the introduction pump 142 in the first direction. As a result, the sample introduced from the culture device 11 into the sample introduction path 130 flows into the sampling path 64 from the connection point 134, passes through the first detection unit 76 and the second detection unit 80 in sequence, and is discharged into the waste liquid container 74. The first detection unit 76 and the second detection unit 80 detect this sample.

[0080] Furthermore, in the main process, it is preferable that the controller 68 controls the rotation speed of the introduction pump 142, similar to the first embodiment. That is, in the first stage after the start of the main process, the controller 68 rotates the introduction pump 142 at a fast first rotation speed to circulate the sample at a first flow rate. In the second stage after the first stage, the controller 68 rotates the introduction pump 142 at a second rotation speed slower than the first rotation speed to circulate the sample at a second flow rate slower than the first flow rate. This enhances the effect of washing away aggregates captured on the filter surface on the sampling path 64 side, while reducing the excessive capture of aggregates on the filter surface on the culture device 11 side (see also Figures 8A and 8B).

[0081] Meanwhile, during the cleaning process, as shown in Figure 11, the controller 68 opens the cleaning solution clamp 94a and the waste liquid clamp 94c while closing the standard solution clamp 94b. The controller 68 also rotates the main mechanism pump 92 while rotating the introduction pump 142 in the second direction. As a result, the cleaning solution in the cleaning solution container 70 flows out from the cleaning solution branching path 71 into the sampling path 64, and is divided into a first cleaning solution that flows directly through the sampling path 64 from the connection point 134, and a second cleaning solution that flows from the connection point 134 into the sample introduction path 130.

[0082] The first washing solution flows through the detection unit 75 (first detection unit 76, second detection unit 80) to wash the multiple first element units 78 and multiple second element units 82. Meanwhile, the second washing solution flows through the sample introduction path 130 and flows into the sample outflow path 54, where it passes through the sterile filter 58. As the second washing solution passes through the sterile filter 58, it removes any aggregates attached to the sterile filter 58. The second washing solution that has passed through the sterile filter 58 flows through the EC circulation circuit 44a of the culture device 11, but the amount is small, and it is also discharged to the waste liquid section 20 via the EC waste liquid circuit 48 as needed.

[0083] In this washing process, it is preferable for the controller 68 to set the rotational speed of the introduction pump 142 in the second direction to be slower than the rotational speed of the main mechanism pump 92. This reduces the amount of second washing solution flowing through the sample introduction path 130 to less than the amount of first washing solution flowing through the sampling path 64. Therefore, the amount of second washing solution flowing into the culture apparatus 11 can be sufficiently reduced. In addition, the controller 68 may also control the rotational speed of the introduction pump 142 during the washing process, similar to the first embodiment.

[0084] As described above, the sampling device 60A can be configured to perform the cleaning process and the deposit removal process simultaneously by reversing the rotation direction of the introduction pump 142. This makes the sampling device 60A (sampling kit 62) even simpler in configuration, improving user workability and handling.

[0085] [Third Embodiment] The sampling device 60B according to the third embodiment differs from the sampling device 60A according to the second embodiment in that, as shown in Figure 12, a bypass route 150 that bypasses the main mechanism pump 92 is connected to the sampling path 64. In addition, a bypass route clamp 152 is provided in the bypass route 150 to open and close the bypass route 150. The bypass route clamp 152 is provided in the main mechanism 90 and opens and closes the flow path of the bypass route 150 under the control of the controller 68.

[0086] By configuring the sampling device 60B as described above, it becomes possible to perform the deposit removal process at a different time than the cleaning process. For example, the controller 68 of the sampling device 60B can transition to the deposit removal process based on the pressure detected by the pressure sensor 146 and the bubbles detected by the bubble sensor 148 when the sampling process is being performed.

[0087] Specifically, during the cleaning process, as shown in Figure 13, the controller 68 opens the cleaning liquid clamp 94a and the waste liquid clamp 94c while closing the standard liquid clamp 94b and the bypass route clamp 152. The controller 68 also rotates the main mechanism pump 92 while stopping the introduction pump 142. As a result, the cleaning liquid in the cleaning liquid storage section 70 flows through the sampling path 64 where the main mechanism pump 92 is located to the first detection section 76 and the second detection section 80.

[0088] In contrast, during the deposit removal process, the controller 68 opens the cleaning solution clamp 94a and the bypass route clamp 152, as shown in Figure 14, while closing the standard solution clamp 94b and the waste liquid clamp 94c. The controller 68 also stops the rotation of the main mechanism pump 92 while rotating the introduction pump 142 in the second direction. As a result, the cleaning solution in the cleaning solution container 70 flows through the sampling route 64 via the bypass route 150 and into the sample introduction route 130. Consequently, the sterile filter 58 provided in the culture device 11 has aggregates removed as the liquid moves from the sample introduction route 130 side to the EC circulation circuit 44a side.

[0089] Thus, by applying the bypass route 150 and the bypass route clamp 152, the sampling device 60B can guide the washing solution to the sterile filter 58 and remove aggregates solely by the rotation of the introduction pump 142. Of course, in this sampling device 60B as well, control may be implemented to change the rotation speed of the introduction pump 142 during the adhering material removal process.

[0090] [Fourth Embodiment] The sampling device 60C according to the fourth embodiment differs from the sampling device 60A described above in that, as shown in Figure 15, it includes an air port 160 that can take in air into the sampling path 64 and an air port clamp 162 that opens and closes the air port 160. In other words, the sampling device 60C is configured to remove aggregates from the sterile filter 58 of the culture device 11 by allowing air to flow in from the air port 160 and pushing back the culture medium in the sampling path 64 and the sample introduction path 130.

[0091] Specifically, the airport 160 is connected to a branching point 161 provided in the sampling path 64 downstream of the detection unit 75 (between the second detection unit 80 and the waste liquid containment unit 74). The airport 160 is equipped with an air filter 164 at an extension end that extends from the sampling path 64. The air filter 164 has the function of blocking the outflow of liquid from the sampling path 64 while allowing air to flow into the sampling path 64.

[0092] The airport clamp 162 is located in the main mechanism 90 and opens and closes the flow path of the airport 160 under the control of the controller 68. In addition, in the sample introduction path 130, similar to the sampling device 60A, a pressure sensor 146 and a bubble sensor 148 are positioned upstream of the introduction pump 142.

[0093] By installing the air filter 164 at the branching point 161, a sufficient amount of liquid can be secured to clean more filters.

[0094] The sampling device 60C according to the fourth embodiment is basically configured as described above, and its operation will be explained below. In the sampling method using the sampling device 60C, the deposit removal step is performed at a different timing than the washing step. The steps other than the deposit removal step are basically the same as those of the sampling device 60A.

[0095] For example, in the sampling process (main process), the controller 68 of the sampling device 60C closes the washing solution clamp 94a, the standard solution clamp 94b, and the air port clamp 162, as shown in Figure 16, while opening the waste liquid clamp 94c. The controller 68 also stops the rotation of the main mechanism pump 92 while rotating the introduction pump 142 in the first direction. In this case, it is preferable that the controller 68 controls the rotation speed of the introduction pump 142, similar to the first embodiment. As a result, the sample introduced from the culture device 11 into the sample introduction path 130 flows into the sampling path 64 from the connection point 134, passes through the first detection unit 76 and the second detection unit 80 in sequence, and is discharged into the waste liquid storage unit 74. The first detection unit 76 and the second detection unit 80 detect this sample.

[0096] Meanwhile, in the deposit removal process, as shown in Figure 17, the controller 68 opens the air port clamp 162 while closing the cleaning solution clamp 94a, the standard solution clamp 94b, and the waste liquid clamp 94c. The controller 68 then stops the rotation of the main mechanism pump 92 and rotates the introduction pump 142 in the second direction. The rotation of the introduction pump 142 creates negative pressure in the sample introduction path 130 and the sampling path 64 downstream of the introduction pump 142. As a result, air flows into the sampling path 64 via the air port 160 (air filter 164).

[0097] The sample (culture medium) remaining in the sampling path 64 and the sample introduction path 130 flows toward the culture device 11 under the rotation of the introduction pump 142 in the second direction. This culture medium passes through the sterile filter 58 in the sample outflow path 54 of the culture device 11 (see also Figure 2), removing any aggregates attached to the sterile filter 58.

[0098] Furthermore, during the adhesion removal process, the controller 68 monitors whether the air flowing from the air port 160 into the sampling path 64 has reached the bubble sensor 148, based on the detection signal from the bubble sensor 148. When the bubble sensor 148 detects air, the controller 68 stops the rotation of the introduction pump 142 and terminates the adhesion removal process. This allows the sampling device 60C to remove aggregates from the sterile filter 58 of the culture device 11 using the culture medium while preventing air from reaching the sterile filter 58. In addition, by not using a washing solution in the adhesion removal process, the sampling device 60C can reliably prevent the washing solution from flowing into the culture device 11.

[0099] It should be noted that the sampling devices 60, 60A to 60C according to the first to fourth embodiments can be adapted to other embodiments by appropriately selecting parts of the configuration of each embodiment. For example, the waste liquid path 138 provided in the sample introduction path 130 of the first embodiment may be adapted to the second to fourth embodiments.

[0100] The technical concepts and effects that can be understood from the above embodiments are described below.

[0101] One aspect of the present invention is a sampling method for collecting a liquid sample from a cell culture section (culture device 11) to a sampling section (sampling devices 60, 60A to 60C), wherein the sampling section comprises a sampling path 64 through which the sample flows, a detection unit 75 provided in the sampling path 64 so as to be in contact with the sample, and a sample introduction path 130 connecting the culture section and the sampling path 64 upstream of the detection unit 75, wherein the culture section or the sample introduction path 130 is equipped with sterile filters 58, 136 in the section up to the introduction of the sample into the sampling path 64, and the sampling step comprises introducing a sample from the culture section to the sampling path 64 via the sample introduction path 130 and detecting the sample with the detection unit 75, and a deposit removal step of removing deposits attached to the sterile filters 58, 136 by flowing fluid from the sampling path 64 to the sample introduction path 130.

[0102] As described above, even with a sampling method equipped with sterile filters 58 and 136, by flowing fluid from the sampling path 64 side during the deposit removal process, deposits attached to the sterile filters 58 and 136 can be removed by performing the sampling process. This improves the clogging of the sterile filters 58 and 136 by deposits. Therefore, the sampling method allows for good sample collection through the sterile filters 58 and 136 while maintaining a sterile state on the culture section (culture device 11) side with the sterile filters 58 and 136.

[0103] Furthermore, the sampling process involves flowing the sample from the culture section (culture device 11) through the sample introduction path 130 to the sampling path 64 at multiple flow rates. This enhances the effect of washing away aggregates captured on the filter surface on the sampling path side, while reducing the excessive capture of aggregates on the filter surface on the culture section side.

[0104] Furthermore, the sampling process involves gradually changing the sample flow rate from fast to slow. This allows for the washing away of aggregates captured on the filter surface on the sampling path side at the start of sampling using a fast flow rate, and then enabling stable sample collection and detection at the subsequent slower flow rate.

[0105] Furthermore, the fast flow rate (first flow rate) is set to a range of 3 to 10 times that of the slow flow rate (second flow rate). This allows the sampling method to more smoothly pass aggregates through the sterile filters 58 and 136.

[0106] Furthermore, the sampling unit (sampling devices 60, 60A~60C) is connected to the upstream side of the sampling path 64 and includes a cleaning liquid storage unit 70 containing cleaning liquid. In the deposit removal process, the cleaning liquid is introduced as a fluid from the sampling path 64 into the sample introduction path 130. As a result, the sampling method can remove deposits from the sterile filters 58 and 136 using the cleaning liquid.

[0107] Furthermore, a cleaning process is performed at a different time from the deposit removal process by circulating cleaning solution from the cleaning solution containment section 70 to the detection section 75 via the sampling path 64 to clean the detection section 75. This allows the sampling method to perform the deposit removal process at an appropriate time.

[0108] Furthermore, the sample introduction path 130 is equipped with at least one of a pressure sensor 146 for detecting the pressure within the sample introduction path 130 and a bubble sensor 148 for detecting bubbles within the sample introduction path 130. In the sampling process, the deposit removal process is performed based on whether the pressure detected by the pressure sensor 146 exceeds a pressure threshold or whether the number of bubbles detected by the bubble sensor 148 exceeds a predetermined level. This allows the sampling method to accurately detect the degree of clogging of the sterile filters 58 and 136, and enables the deposit removal process to be performed at the necessary timing.

[0109] Furthermore, a waste liquid route 138, which is a different route from the culture section, is connected to the sample introduction route 130 between the culture section (culture device 11) and the sterile filter 136. In the deposit removal process, the fluid that has passed through the sterile filter 136 is guided to the waste liquid route 138. As a result, even if cleaning solution is introduced into the sample introduction route 130 to remove deposits from the sterile filter 136, the cleaning solution is discharged via the waste liquid route 138, preventing it from flowing into the culture device 11.

[0110] Furthermore, the sampling path 64 includes a first pump (main mechanism side pump 92) that circulates the cleaning solution from the cleaning solution containment section 70 to the sampling path 64, and a bypass path 150 that bypasses the first pump. The sample introduction path 130 includes a second pump (introduction pump 142) that can rotate in a first direction for introducing the sample into the sampling path 64 and a second direction for directing the fluid toward the culture section (culture device 11). The first pump is rotated, and the bypass path 150 is blocked to circulate the cleaning solution to the detection section 75. In the deposit removal process, the rotation of the first pump is stopped, and the second pump is rotated in the second direction, causing the cleaning solution to flow into the sample introduction path 130 via the bypass path 150. This makes it easy to switch between the cleaning process and the deposit removal process in the sampling method.

[0111] Furthermore, a cleaning process is performed simultaneously with the deposit removal process by circulating cleaning solution from the cleaning solution containment section 70 to the detection section 75 via the sampling path 64 to clean the detection section 75. This allows the sampling method to efficiently perform cleaning of the detection section 75 with cleaning solution and removal of deposits attached to the sterile filter 58.

[0112] Furthermore, the amount of cleaning solution flowing through the sample introduction path 130 in the deposit removal process is set to be less than the amount of cleaning solution flowing through the detection unit 75 in the cleaning process. This allows the sampling method to remove deposits from the sterile filter 58 while stably cleaning the detection unit 75.

[0113] Furthermore, the sampling unit (sampling device 60C) is connected to the sampling path 64 downstream of the detection unit 75, and the sampling path 64 is equipped with an air port 160 that can take in air. In the deposit removal process, the sample in the sampling path 64 is circulated through the sample introduction path 130 using air taken in from the air port 160, thereby removing the deposits with the sample. In this way, the sampling method can also remove deposits from the sterile filter 58 by causing the sample to flow backward with the inflow of air.

Claims

1. A sampling method for collecting a liquid sample from a cell culture section to a sampling section, The sampling unit is The sampling route through which the aforementioned sample is distributed, A detection unit is provided in the sampling path so as to be in contact with the sample, The system includes a sample introduction path connecting the culture section and the sampling path upstream of the detection section, The culture section or the sample introduction route is equipped with a sterile filter in the section up to the introduction of the sample into the sampling route. The sample introduction path is equipped with a pump for circulating the fluid within the sample introduction path. A sampling step in which the sample is introduced from the culture unit to the sampling path via the sample introduction path, and the sample is detected by the detection unit, The system includes a deposit removal step, which involves flowing the fluid from the sampling path to the sample introduction path to remove deposits adhering to the sterile filter during the sampling step, In the sampling step, the rotational speed of the pump is changed to allow the sample to flow from the culture section through the sample introduction path to the sampling path at multiple flow rates. Sampling method.

2. In the sampling method according to claim 1, In the sampling step, the multiple flow velocities of the sample are changed in steps from a fast flow velocity to a slow flow velocity. Sampling method.

3. In the sampling method according to claim 2, The aforementioned high flow velocity is set to be in the range of 3 to 10 times the aforementioned slow flow velocity. Sampling method.

4. In the sampling method according to any one of claims 1 to 3, The sampling unit is connected to the upstream side of the sampling path and includes a cleaning liquid storage unit that contains cleaning liquid. In the deposit removal step, the cleaning solution is introduced as the fluid from the sampling path into the sample introduction path. Sampling method.

5. In the sampling method according to claim 4, A cleaning step, in which the cleaning liquid is circulated from the cleaning liquid storage section to the detection section via the sampling path to clean the detection section, is performed at a different time than the deposit removal step. Sampling method.

6. In the sampling method according to claim 5, The sample introduction path is provided with at least one of the following: a pressure sensor for detecting the pressure within the sample introduction path, and a bubble sensor for detecting bubbles within the sample introduction path. Based on the sampling step, if the pressure detected by the pressure sensor becomes equal to or above a pressure threshold, or if the number of bubbles detected by the bubble sensor becomes equal to or above a predetermined level, the deposit removal step is performed. Sampling method.

7. In the sampling method according to any one of claims 4 to 6, A wastewater pathway, which is a different pathway from the culture pathway, is connected to the sample introduction pathway between the culture section and the sterile filter. In the deposit removal step, the fluid that has passed through the sterile filter is guided to the wastewater path. Sampling method.

8. In the sampling method according to any one of claims 4 to 7, The sampling path comprises a first pump that circulates the cleaning liquid from the cleaning liquid containment unit to the sampling path, and a bypass path that bypasses the first pump. The sample introduction path includes a second pump that is rotatable in a first direction for introducing the sample into the sampling path and a second direction for directing the fluid toward the culture section. The first pump is rotated, and the bypass path is blocked, allowing the cleaning fluid to flow to the detection unit. In the deposit removal step, the rotation of the first pump is stopped, and the second pump is rotated in the second direction, thereby causing the cleaning solution to flow into the sample introduction path via the detour path. Sampling method.

9. In the sampling method according to claim 4, A cleaning process is performed simultaneously with the deposit removal process, in which the cleaning liquid is circulated from the cleaning liquid storage section to the detection section via the sampling path to clean the detection section. Sampling method.

10. In the sampling method according to claim 9, The amount of cleaning solution flowing through the sample introduction path in the deposit removal step is made less than the amount of cleaning solution flowing through the detection unit in the cleaning step. Sampling method.

11. In the sampling method according to any one of claims 1 to 4, The sampling unit is connected to the sampling path downstream of the detection unit and is equipped with an air port capable of taking in air into the sampling path. In the deposit removal step, the sample in the sampling path is circulated through the sample introduction path using air taken in from the air port, thereby removing the deposit with the sample. Sampling method.

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