Sampling method

Abstract

A sampling method comprising: a sampling step for passing a sample through a sampling flow path (30) from a cell culture device (200) and measuring the concentration of a specific component in the sample by means of a second sensor (28) and a gas concentration sensor (62); a washing step for passing a wash solution through the sampling flow path (30) and passing the wash solution through the second sensor (28) and the gas concentration sensor (62) subsequent to the sampling step; and an air introduction step for replacing the wash solution remaining in the inside of the gas concentration sensor (62) with air subsequent to the washing step.

Description

[Technical Field]

[0001] The present invention relates to a sampling method. [Background technology]

[0002] For example, US Pat. No. 9,442,047 discloses a sampling method using a sampling system having a sampling channel for collecting a liquid sample from a cell culture device. Summary of the Invention

[0003] Incidentally, a sampling system may include a biosensor and a gas concentration sensor provided in a sampling flow path so as to come into contact with the sample, and an inlet path for introducing a cleaning solution into the sampling flow path upstream of the biosensor and the gas concentration sensor. In this case, for example, a sampling process is performed in which the sample is circulated through the sampling flow path to bring the sample into contact with the biosensor and the gas concentration sensor, and a cleaning process is performed in which the cleaning solution is introduced from the inlet path into the sampling flow path to remove the sample adhering to the biosensor in order to prevent deterioration of the catalyst in the biosensor.

[0004] After the cleaning process is completed, cleaning liquid remains inside the gas concentration sensor. If the internal volume of the gas concentration sensor is relatively large, the cleaning liquid may get mixed into the sample inside the gas concentration sensor during the sampling process. This reduces the measurement accuracy of the gas concentration sensor.

[0005] The present invention has been made in consideration of these problems, and aims to provide a sampling method that can remove sample adhering to a biosensor and prevent a decrease in the measurement accuracy of a gas concentration sensor.

[0006] One aspect of the present invention is a sampling method using a sampling system having a sampling flow path for collecting a sample of a liquid from a cell culture device, wherein the sampling flow path is provided with a biosensor and a gas concentration sensor so as to come into contact with the sample, and the sampling method includes: a sampling step of circulating the sample from the cell culture device to the sampling flow path and measuring the concentration of a predetermined component in the sample with the biosensor and the gas concentration sensor; a cleaning step of circulating a cleaning liquid through the sampling flow path after the sampling step to circulate the cleaning liquid through the biosensor and the gas concentration sensor; and an air introduction step of replacing the cleaning liquid remaining inside the gas concentration sensor with air after the cleaning step, wherein the sampling step is performed two or more times, and the second and subsequent sampling steps are performed after the air introduction step.

[0007] According to the present invention, since the cleaning process is performed after the sampling process, the sample adhering to the biosensor during the sampling process can be removed with a cleaning solution in the cleaning process. Furthermore, in the air introduction process after the cleaning process, the cleaning solution remaining inside the gas concentration sensor is replaced with air. Therefore, in the sampling process after the cleaning process, it is possible to prevent the cleaning solution from mixing with the sample inside the gas concentration sensor. Therefore, it is possible to prevent a decrease in the measurement accuracy of the gas concentration sensor. [Brief explanation of the drawings]

[0008] [Figure 1] 1 is a schematic configuration diagram of a sampling system according to an embodiment of the present invention. [Figure 2] FIG. 2 is a diagram illustrating the configuration of a main part of the cell culture device. [Figure 3] 2 is a flowchart illustrating a sampling method using the sampling system of FIG. 1. [Figure 4] 4 is a flowchart for explaining the sampling process of FIG. 3. [Figure 5] FIG. 10 is a diagram illustrating a first operation of the sampling method. [Figure 6] FIG. 10 is a diagram illustrating a second operation of the sampling method. [Figure 7] FIG. 10 is a diagram illustrating a third operation of the sampling method. [Figure 8] FIG. 4 is a diagram illustrating a fourth operation of the sampling method. [Figure 9] FIG. 5 is a diagram illustrating a fifth operation of the sampling method. DETAILED DESCRIPTION OF THE INVENTION

[0009] A preferred embodiment of the sampling method according to the present invention will be described below with reference to the accompanying drawings.

[0010] 1, a sampling system 10 according to one embodiment of the present invention collects liquid samples from a plurality of cell culture devices 200 and measures the concentration of a predetermined component in the sample. The sampling system 10 includes a sampling kit 12, a circuit control device 14 to which the sampling kit 12 can be detachably attached, and a controller 16. The sampling kit 12 is a disposable item, and the circuit control device 14 is a reusable item.

[0011] In this embodiment, a first cell culture device 200A and a second cell culture device 200B are connected to the sampling kit 12 as a plurality of cell culture devices 200. As shown in Fig. 2, the cell culture device 200 has a bioreactor 202 for culturing cells. The cells to be cultured are those separated from biological tissue, and for example, cells contained in blood (such as T cells) or stem cells (such as ES cells, iPS cells, mesenchymal stem cells) are used.

[0012] The bioreactor 202 is configured as a so-called hollow fiber bioreactor. The bioreactor 202 includes a large number (plurality) of hollow fibers 204 and a cylindrical housing 206 that houses these hollow fibers 204. A plurality of pores (not shown) are formed in the wall of the hollow fibers 204. The pores communicate between an IC (intra capillary) region, which is the lumen of the hollow fibers 204, and an EC (extra capillary) region located outside the hollow fibers 204 within the housing 206. The diameter of the pores is set to a size that prevents the passage of macromolecules (such as cells) while allowing the passage of small molecules (e.g., water, ions, oxygen, lactate, etc.).

[0013] Housing 206 is provided with an IC inlet port 208, an IC outlet port 210, an EC inlet port 212, and an EC outlet port 214. IC inlet port 208 is provided at one end of housing 206. IC inlet port 208 introduces liquid (such as a cell-containing solution or culture medium) guided from IC inlet channel 216 into the IC region of bioreactor 202. IC outlet port 210 is provided at the other end of housing 206. IC outlet port 210 guides liquid that has circulated through the IC region of bioreactor 202 to IC outlet channel 218.

[0014] The EC inlet port 212 and the EC outlet port 214 are provided on the outer peripheral surface of the housing 206. The EC inlet port 212 introduces the culture medium guided from the EC inlet channel 220 into the EC region of the bioreactor 202. The EC outlet port 214 guides the culture medium that has circulated through the EC region of the bioreactor 202 to the EC outlet channel 222. The culture medium may be selected appropriately depending on the cells of the living organism, and for example, a medium prepared by adding various amino acids, vitamins, serum, etc. to a balanced salt solution (BSS) as a base solution is used.

[0015] A connection line 224 is connected to the EC outlet channel 222 to guide the culture medium that has flowed through the EC region to the sampling kit 12. The connection line 224 is provided with a sterile filter 226 and a sampling connector 228. The sterile filter 226 keeps the portion of the cell culture device 200 that is closer to the EC outlet channel 222 than the sterile filter 226 sterile. The introduction connector 42 of the sampling kit 12 is detachable from the sampling connector 228.

[0016] In this embodiment, the sampling system 10 collects, as a sample, the culture medium that has flowed through the EC region of the cell culture device 200. However, the sample collected by the sampling system 10 is not limited to the culture medium that has flowed through the EC region, and may be the culture medium that has flowed through the IC region or other liquids.

[0017] In FIG. 1, the sampling kit 12 includes a cleaning liquid storage section 18, a standard liquid storage section 20, a waste liquid storage section 22, a connection circuit 24, a first sensor 26, and a second sensor .

[0018] The cleaning liquid storage section 18, the standard liquid storage section 20, and the waste liquid storage section 22 are each formed into a bag shape and made of a flexible material such as soft resin, for example, polyvinyl chloride or polyolefin. However, the cleaning liquid storage section 18, the standard liquid storage section 20, and the waste liquid storage section 22 can be modified as needed as long as they are capable of storing liquids.

[0019] A cleaning liquid is stored in the cleaning liquid storage section 18. A buffer solution or physiological saline is used as the cleaning liquid. Examples of the buffer solution include PBS (Phosphate Buffered Saline) and TBS (Tris-Buffered Saline). However, the cleaning liquid is not limited to those mentioned above.

[0020] A standard solution is stored in the standard solution storage section 20. The standard solution is a liquid for calibrating the first sensor 26 and the second sensor 28. Specifically, the standard solution is a liquid in which the pH value, O2 value (oxygen concentration), CO2 value (carbon dioxide concentration), glucose value (glucose concentration), and lactic acid value (lactic acid concentration) are set to specified values.

[0021] The waste liquid storage section 22 is for storing waste liquids (sample, cleaning liquid, and standard liquid) that have flowed through the connection circuit 24. The waste liquid storage section 22 is an empty bag that does not contain any liquid before the sampling kit 12 is used.

[0022] The connection circuit 24 has a sampling flow path 30 for collecting a sample from the cell culture device 200, an introduction path 32 for introducing a cleaning solution to the sampling flow path 30, a standard solution introduction path 33 for introducing a standard solution to the introduction path 32, and an air introduction path 35 for introducing air to the sampling flow path 30. The sampling flow path 30 includes a first sample introduction path 34a, a second sample introduction path 34b, and a sample flow path 36.

[0023] The first sample introduction path 34a introduces the sample (culture medium) of the first cell culture device 200A into the sample flow path 36. One end of the first sample introduction path 34a is provided with an introduction connector 42 that is attached to the sampling connector 228 of the first cell culture device 200A (see FIG. 2). The other end of the first sample introduction path 34a is connected to one end of the sample flow path 36. Hereinafter, the connection portion between the first sample introduction path 34a and the sample flow path 36 will be referred to as a first connection portion 38.

[0024] The second sample introduction path 34b introduces the sample (culture medium) of the second cell culture device 200B into the sample flow path 36. One end of the second sample introduction path 34b is provided with an introduction connector 42 that is attached to the sampling connector 228 of the second cell culture device 200B (see FIG. 2). The other end of the second sample introduction path 34b is connected to a midpoint of the sample flow path 36. Hereinafter, the connection portion between the second sample introduction path 34b and the sample flow path 36 will be referred to as a second connection portion 40.

[0025] The sample flow path 36 includes an intermediate flow path 44 that connects the first connecting portion 38 and the second connecting portion 40 to each other, and a sensor flow path 46 that connects the second connecting portion 40 and the waste liquid storage portion 22 to each other.

[0026] One end of the introduction path 32 is connected to the cleaning liquid storage section 18. The other end of the introduction path 32 is connected to the first connecting section 38. One end of the standard solution introduction path 33 is connected to the standard solution storage section 20. The other end of the standard solution introduction path 33 is connected to an intermediate portion of the introduction path 32. Hereinafter, the connecting section between the introduction path 32 and the standard solution introduction path 33 will be referred to as the third connecting section 48.

[0027] An air port 50 open to the atmosphere and a sterile filter 52 are provided at one end of the air introduction path 35. The sterile filter 52 maintains the connection circuit 24 in a sterile state. The other end of the air introduction path 35 is connected to a section of the sensor flow path 46 between the second connecting part 40 and the first sensor 26. Hereinafter, the connecting part between the sensor flow path 46 and the air introduction path 35 will be referred to as a fourth connecting part 54.

[0028] The first sensor 26 and the second sensor 28 are provided in the sensor flow path 46 so as to come into contact with the sample. The first sensor 26 is an integrally molded product and includes a pH sensor 60 and a gas concentration sensor 62. The pH sensor 60 measures the pH of the sample. The gas concentration sensor 62 measures the gas concentration of the sample. Specifically, the gas concentration sensor 62 includes an O2 sensor 64 that measures the O2 concentration of the sample and a CO2 sensor 66 that measures the CO2 concentration of the sample.

[0029] The second sensor 28 is, for example, a biosensor such as an enzyme sensor. The second sensor 28 is provided downstream of the first sensor 26 in the sensor flow path 46. The second sensor 28 is an integrally molded product and includes a glucose sensor 68 that measures the glucose concentration in a sample and a lactate sensor 70 that measures the lactate concentration in the sample. The second sensor 28 is not limited to an enzyme sensor and may include a non-enzymatic glucose sensor. Furthermore, the measurement items of the second sensor 28 are not limited to glucose and lactate but may include glutamic acid, etc.

[0030] The circuit control device 14 includes a plurality of clamps 72 and one pump 74. In this embodiment, the circuit control device 14 includes the plurality of clamps 72, which include a first clamp 72a (first opening / closing unit), a second clamp 72b, a third clamp 72c, a fourth clamp 72d (second opening / closing unit), a fifth clamp 72e, and a sixth clamp 72f (third opening / closing unit).

[0031] The first clamp 72a is positioned to face the first sample introduction path 34a when the sampling kit 12 is attached to the circuit control device 14 (hereinafter referred to as the "set state") and opens and closes the internal flow path of the first sample introduction path 34a. The second clamp 72b is positioned to face the second sample introduction path 34b when the sampling kit 12 is set and opens and closes the internal flow path of the second sample introduction path 34b. The third clamp 72c is positioned to face a portion of the sensor flow path 46 between the second sensor 28 and the waste liquid storage section 22 when the sampling kit 12 is set and opens and closes the internal flow path of that portion of the sensor flow path 46. The fourth clamp 72d is positioned to face a portion of the introduction path 32 upstream of the third connecting section 48 when the sampling kit 12 is set and opens and closes the internal flow path of that portion of the introduction path 32. The fifth clamp 72e is positioned to face the standard solution introduction path 33 when the sampling kit 12 is set and opens and closes the internal flow path of the standard solution introduction path 33. In the set state, the sixth clamp 72f is disposed so as to face the air introduction path 35, and opens and closes the internal flow path of the air introduction path 35.

[0032] The pump 74 applies a flow force to the liquid inside by rotating in a manner that squeezes the wall that constitutes the flow path (tube) of the connection circuit 24. When set, the pump 74 is disposed so as to come into contact with a portion of the sensor flow path 46 between the second connecting portion 40 and the first sensor 26. The pump 74 performs a first rotation operation (a rotation operation in the direction of arrow R1) so as to apply a flow force to the liquid flowing through the sensor flow path 46 in a direction toward the first sensor 26 (waste liquid storage portion 22). The pump 74 may also perform a second rotation operation (a rotation operation in the direction of arrow R2) so as to apply a flow force to the liquid flowing through the sensor flow path 46 in a direction toward the second connecting portion 40.

[0033] The controller 16 (control unit) is a computer having a processor, memory, and input / output interface (not shown). The controller 16 performs overall control of the entire system by having the processor execute a program stored in the memory. The controller 16 is connected to the circuit control device 14 by a communication means such as a wired, wireless, network, or a combination thereof. Specifically, the controller 16 controls the operation of the multiple clamps 72 and the pump 74.

[0034] Next, a sampling method using the sampling system 10 will be described.

[0035] As shown in FIG. 3, the sampling method includes a preparation step, a priming step, an air introduction step, a sampling step, a cleaning step, and a calibration step.

[0036] First, in the preparation process (step S1), as shown in Figures 1 and 2, the sampling kit 12 is attached (set) to the circuit control device 14, and the introduction connector 42 of the first sample introduction path 34a is connected to the sampling connector 228 of the first cell culture device 200A, and the introduction connector 42 of the second sample introduction path 34b is connected to the sampling connector 228 of the second cell culture device 200B.

[0037] Subsequently, in the priming step (step S2 in FIG. 3), as shown in FIG. 5, the controller 16 opens the third clamp 72c and the fourth clamp 72d and closes the first clamp 72a, the second clamp 72b, the fifth clamp 72e, and the sixth clamp 72f, and causes the pump 74 to perform a first rotation operation. Then, the cleaning liquid in the cleaning liquid storage section 18 is guided by the action of the pump 74 from the introduction path 32 through the first connecting section 38, the intermediate flow path 44, the second connecting section 40, and the sensor flow path 46 to the waste liquid storage section 22.

[0038] Thereafter, an air introduction step (step S3 in FIG. 3) is performed. Specifically, as shown in FIG. 6, controller 16 opens third clamp 72c and sixth clamp 72f and closes first clamp 72a, second clamp 72b, fourth clamp 72d, and fifth clamp 72e, and causes pump 74 to perform a first rotation operation. This causes air introduced into air introduction path 35 from air port 50 via sterile filter 52 to be guided to waste liquid storage section 22 via first sensor 26 and second sensor 28 of sensor flow path 46. This forces the cleaning liquid remaining inside first sensor 26 into waste liquid storage section 22.

[0039] The air introduction step is not limited to the example in which the air introduced into the air introduction path 35 is guided to the waste liquid storage section 22. The air introduction step only needs to be able to replace the cleaning liquid remaining inside the gas concentration sensor 62 with air. In other words, the timing at which the introduction of air is stopped (the operation of the pump 74 is stopped) in the air introduction step may be the point in time when air is introduced into the section of the sensor flow path 46 between the first sensor 26 and the second sensor 28.

[0040] Then, the sampling step (step S4 in FIG. 3) is performed. Specifically, the controller 16 selects a sample to be collected (step S5 in FIG. 4). That is, the controller 16 selects whether to collect a sample from the first cell culture device 200A (first sample) or a sample from the second cell culture device 200B (second sample) based on the state of cell culture in the cell culture device 200.

[0041] When the controller 16 selects to collect a first sample, a first sample introduction step is performed (step S6 in FIG. 4). That is, as shown in FIG. 7, the controller 16 opens the first clamp 72a and the third clamp 72c and closes the second clamp 72b, the fourth clamp 72d, the fifth clamp 72e, and the sixth clamp 72f, and causes the pump 74 to perform a first rotation operation. Then, the first sample in the first cell culture device 200A is guided by the action of the pump 74 to the waste liquid storage section 22 via the first sample introduction channel 34a, the first connecting section 38, the intermediate flow path 44, the second connecting section 40, and the sensor flow path 46.

[0042] At this time, since no cleaning liquid remains inside the first sensor 26 (the cleaning liquid inside the first sensor 26 is replaced with air in the air introduction process), the cleaning liquid does not get mixed into the first sample flowing through the first sensor 26.

[0043] Furthermore, the first sensor 26 and the second sensor 28 come into contact with the first sample. The first sensor 26 measures the pH, O2 concentration, and CO2 concentration in the first sample. The measurement results of the first sensor 26 are transmitted to the controller 16. The second sensor 28 measures the glucose concentration and lactate concentration in the first sample. The measurement results of the second sensor 28 are transmitted to the controller 16. The controller 16 controls the culture conditions of the first cell culture device 200A based on the measurement results of the first sensor 26 and the second sensor 28.

[0044] When the controller 16 selects to collect a second sample, a second sample introduction step is performed (step S7 in FIG. 4). That is, as shown in FIG. 8, the controller 16 opens the second clamp 72b and the third clamp 72c and closes the first clamp 72a, the fourth clamp 72d, the fifth clamp 72e, and the sixth clamp 72f, and causes the pump 74 to perform the first rotation. Then, the second sample in the second cell culture device 200B is introduced into the waste liquid storage section 22 via the second sample introduction path 34b, the second connection section 40, and the sensor flow path 46 by the action of the pump 74.

[0045] At this time, since no cleaning liquid remains inside the first sensor 26 (the cleaning liquid inside the first sensor 26 is replaced with air in the air introduction process), the cleaning liquid does not get mixed into the second sample flowing through the first sensor 26.

[0046] Furthermore, the first sensor 26 and the second sensor 28 come into contact with the second sample. The first sensor 26 measures the pH, O2 concentration, and CO2 concentration in the second sample. The measurement results of the first sensor 26 are transmitted to the controller 16. The second sensor 28 measures the glucose concentration and lactate concentration in the second sample. The measurement results of the second sensor 28 are transmitted to the controller 16. The controller 16 controls the culture conditions of the second cell culture device 200B based on the measurement results of the first sensor 26 and the second sensor 28.

[0047] 3, the controller 16 determines whether or not cell culture has been completed in the first cell culture device 200A and the second cell culture device 200B (step S8). If the controller 16 determines that cell culture has not been completed (step S8: NO), a cleaning step (step S9) is performed. In the cleaning step, the controller 16 operates the multiple clamps 72 and the pump 74 in the same manner as in the priming step, as shown in FIG. 5. Then, the cleaning liquid in the cleaning liquid storage section 18 flows through the first sensor 26 and the second sensor 28 and is guided to the waste liquid storage section 22.

[0048] As a result, in the first sensor 26, the cleaning fluid removes samples adhering to the pH sensor 60, O2 sensor 64, and CO2 sensor 66. In the second sensor 28, the cleaning fluid removes samples adhering to the glucose sensor 68 and lactate sensor 70.

[0049] Thereafter, a calibration process (step S10 in FIG. 3) is performed as necessary. In the calibration process, as shown in FIG. 9, the controller 16 opens the third clamp 72c and the fifth clamp 72e and closes the first clamp 72a, the second clamp 72b, the fourth clamp 72d, and the sixth clamp 72f, and causes the pump 74 to perform a first rotation. Then, the standard solution in the standard solution storage section 20 is guided by the action of the pump 74 to the waste liquid storage section 22 via the standard solution inlet channel 33, the inlet channel 32, the first connecting section 38, the intermediate flow path 44, the second connecting section 40, and the sensor flow path 46.

[0050] At this time, the first sensor 26 measures the pH, O2 concentration, and CO2 concentration in the standard solution. The measurement results of the first sensor 26 are transmitted to the controller 16. The controller 16 calibrates the pH sensor 60, O2 sensor 64, and CO2 sensor 66 based on the measurement results of the first sensor 26. The second sensor 28 measures the glucose concentration and lactate concentration in the standard solution. The measurement results of the second sensor 28 are transmitted to the controller 16. The controller 16 calibrates the glucose sensor 68 and lactate sensor 70 based on the measurement results of the second sensor 28. Once the calibration process is completed, step S3 and subsequent steps are performed sequentially. In this embodiment, the sampling process is performed two or more times.

[0051] When the controller 16 determines that the cell culture has ended (step S8 in FIG. 3: YES), the series of operation flows ends.

[0052] This embodiment has the following advantages.

[0053] According to this embodiment, the cleaning step is performed after the sampling step, so that the sample adhering to the second sensor 28 during the sampling step can be removed with a cleaning liquid in the cleaning step. Furthermore, in the air introduction step after the cleaning step, the cleaning liquid remaining inside the gas concentration sensor 62 is replaced with air. Therefore, in the sampling step after the cleaning step, it is possible to prevent the cleaning liquid from mixing with the sample inside the gas concentration sensor 62. Therefore, it is possible to prevent a decrease in the measurement accuracy of the gas concentration sensor 62.

[0054] The sampling system 10 includes an inlet channel 32 that introduces a cleaning liquid upstream of the second sensor 28 into the sampling flow path 30, an air inlet channel 35 that is connected to the sampling flow path 30 upstream of the gas concentration sensor 62 and is open to the atmosphere, a first clamp 72a that opens and closes the sampling flow path 30 upstream of a first connecting portion 38 with the inlet channel 32, a fourth clamp 72d that opens and closes the inlet channel 32, a sixth clamp 72f that opens and closes the air inlet channel 35, and a pump 74 provided in the sampling flow path 30. In the air introduction step, the pump 74 is driven with the first clamp 72a and the fourth clamp 72d closed and the sixth clamp 72f open, thereby introducing air from the air inlet channel 35 into the sampling flow path 30.

[0055] According to this method, air can be introduced from the air introduction passage 35 to the gas concentration sensor 62 with a simple configuration and control.

[0056] The air introduction path 35 is provided with a sterile filter 52. In the air introduction step, air that has passed through the sterile filter 52 is introduced into the sampling flow path 30.

[0057] In this manner, the sterile filter 52 keeps the sample sterile.

[0058] The sampling flow path 30 has a sample flow path 36 that is provided with a second sensor 28 and a gas concentration sensor 62 and that forms a downstream side of a first connecting portion 38 with the inlet path 32 in the sampling flow path 30, a first sample inlet path 34a that introduces the first sample of the first cell culture device 200A to the first connecting portion 38, and a second sample inlet path 34b that introduces the second sample of the second cell culture device 200B to the upstream side of the second sensor 28 and the gas concentration sensor 62 in the sample flow path 36. The sampling step includes a first sample introduction step of introducing the first sample of the first cell culture device 200A from the first sample introduction path 34a into the sample flow path 36, and a second sample introduction step of introducing the second sample of the second cell culture device 200B from the second sample introduction path 34b into the sample flow path 36.

[0059] According to this method, contamination of the first sample or the second sample inside the gas concentration sensor 62 with the cleaning liquid can be suppressed.

[0060] The sampling system 10 may collect a sample from one cell culture device 200 and measure the concentration of a predetermined component therein. In this case, the sampling system 10 may not be provided with the second sample introduction path 34b. Alternatively, the sampling system 10 may collect samples from three or more cell culture devices 200 individually and measure the concentration of a predetermined component therein. In other words, the number of cell culture devices 200 connected to the sampling flow path 30 may be three or more. In this case, the number of sample introduction paths provided corresponds to the number of cell culture devices 200. The air introduction path 35 may be connected to any of the intermediate flow path 44, the first sample introduction path 34a, the second sample introduction path 34b, and the introduction path 32.

[0061] The present invention is not limited to the above-described embodiment, and various modifications are possible without departing from the spirit of the present invention. In the above-described embodiment, a cell culture system in which the sampling system 10 and the cell culture device 200 are configured separately is shown, but the cell culture system may be one in which the sampling system 10 and the cell culture device 200 are integrated (united).

[0062] The above embodiments can be summarized as follows.

[0063] The above embodiment discloses a sampling method using a sampling system (10) having a sampling flow path (30) for collecting a liquid sample from a cell culture device (200), in which a biosensor (28) and a gas concentration sensor (62) are provided in the sampling flow path so as to come into contact with the sample, and the sampling method includes: a sampling step of circulating the sample from the cell culture device to the sampling flow path and measuring the concentration of a predetermined component in the sample with the biosensor and the gas concentration sensor; a cleaning step of circulating a cleaning liquid through the sampling flow path to circulate the cleaning liquid through the biosensor and the gas concentration sensor after the sampling step; and an air introduction step of replacing the cleaning liquid remaining inside the gas concentration sensor with air after the cleaning step, in which the sampling step is performed two or more times, and the second and subsequent sampling steps are performed after the air introduction step.

[0064] In the above sampling method, the sampling system comprises an inlet path (32) that introduces the cleaning solution upstream of the biosensor in the sampling flow path, an air inlet path (35) that is connected to the sampling flow path upstream of the gas concentration sensor and is open to the atmosphere, a first opening / closing unit (72a) that opens and closes the sampling flow path upstream of the connection unit (38) with the inlet path, a second opening / closing unit (72d) that opens and closes the inlet path, a third opening / closing unit (72f) that opens and closes the air inlet path, and a pump (74) provided in the sampling flow path, and in the air introduction step, the air may be introduced from the air inlet path to the sampling flow path by driving the pump with the first opening / closing unit and the second opening / closing unit closed and the third opening / closing unit open.

[0065] In the above sampling method, a sterile filter (52) may be provided in the air introduction path, and in the air introduction step, the air that has passed through the sterile filter may be introduced into the sampling flow path.

[0066] In the above sampling method, the sampling system has an inlet path that introduces the cleaning solution into the sampling flow path upstream of the biosensor, and the sampling flow path has a sample flow path (36) that forms the downstream side of the connection part with the inlet path in the sampling flow path and in which the biosensor and the gas concentration sensor are provided, a first sample inlet path (34a) that introduces a first sample from a first cell culture device (200A) that is the cell culture device to the connection part, and a second sample inlet path (34b) that introduces a second sample from a second cell culture device (200B) to the upstream side of the biosensor and the gas concentration sensor in the sample flow path, and the sampling process may include a first sample introduction process of introducing the first sample from the first cell culture device from the first sample inlet path into the sample flow path, and a second sample introduction process of introducing the second sample from the second cell culture device from the second sample inlet path into the sample flow path.

Claims

1. A sampling method using a sampling system having a sampling flow path for collecting a sample of a liquid in a cell culture device, comprising: a biosensor and a gas concentration sensor are provided in the sampling flow path so as to come into contact with the sample, and a waste liquid storage section is also provided; the biosensor is provided downstream of the gas concentration sensor; the waste liquid storage section is provided downstream of the biosensor, a sampling step of passing the sample from the cell culture device through the sampling flow path and measuring the concentration of a predetermined component in the sample with the biosensor and the gas concentration sensor; a cleaning step of circulating a cleaning liquid through the sampling flow path after the sampling step, and causing the cleaning liquid to flow through the biosensor and the gas concentration sensor; an air introducing step of replacing the cleaning liquid remaining inside the gas concentration sensor with air after the cleaning step, The sampling step is carried out two or more times, The second or subsequent sampling step is performed after the air introduction step, In the air introduction step, the air is introduced into the waste liquid storage section via the gas concentration sensor and the biosensor, thereby pushing out the cleaning liquid remaining inside the biosensor into the waste liquid storage section, a sampling method.

2. 2. The sampling method according to claim 1, The sampling system comprises: an introduction path for introducing the cleaning solution into the sampling flow path upstream of the biosensor; an air introduction passage connected to the sampling passage upstream of the gas concentration sensor and open to the atmosphere; a first opening / closing unit that opens and closes a portion of the sampling flow path upstream of a connection portion with the introduction path; a second opening / closing unit that opens and closes the introduction path; a third opening / closing unit that opens and closes the air introduction path; a pump provided in the sampling flow path, In the air introduction step, the air is introduced from the air introduction path to the sampling flow path by driving the pump while closing the first opening / closing unit and the second opening / closing unit and opening the third opening / closing unit.

3. 3. The sampling method according to claim 2, The air introduction path is provided with a sterile filter, In the air introducing step, the air that has passed through the sterile filter is introduced into the sampling flow path.

4. 2. The sampling method according to claim 1, the sampling system has an introduction path for introducing the cleaning solution into the sampling flow path upstream of the biosensor, The sampling flow path is a sample flow path that forms a downstream side of a connecting portion with the introduction path in the sampling flow path and in which the biosensor and the gas concentration sensor are provided; a first sample introduction path that introduces a first sample from a first cell culture device that is the cell culture device into the connecting portion; a second sample introduction path that introduces a second sample from a second cell culture device to a portion of the sample flow path upstream of the biosensor and the gas concentration sensor; In the sampling step, a first sample introduction step of introducing the first sample of the first cell culture device from the first sample introduction path into the sample flow path; A sampling method comprising: a second sample introduction step of introducing the second sample of the second cell culture device from the second sample introduction path into the sample flow path.

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