Simulation device, simulation system, and simulation method

Abstract

The simulation device 14 includes an acquisition unit 140 that acquires the disappearance rate at which the protein is disappeared regardless of the cells, the consumption rate at which the cells consume the protein under first culture conditions, and condition data indicating second culture conditions different from the first culture conditions, a simulation execution unit 142 that simulates the change in protein concentration associated with cell proliferation under the second culture conditions using the disappearance rate, consumption rate, and the condition data, and a display unit 134 that acquires the protein concentration under the second culture conditions as a result of the simulation, and displays whether the protein concentration falls within a predetermined range.

Description

[Technical field]

[0001] The present invention relates to a simulation device, a simulation system, and a simulation method for simulating cell proliferation in a cell culture device. [Background technology]

[0002] JP 2020-171241 A shows a cell culture device. The cell culture device includes a bioreactor, a supply unit, a recovery unit, and a plurality of flow paths. Some of the flow paths form a circulation path together with the bioreactor. The supply unit supplies a cell-containing liquid and a culture medium (culture liquid) to the bioreactor. The bioreactor cultures cells. During cell culture, a portion of the culture medium (first culture medium) circulates through the circulation path. During cell culture, a portion of the culture medium (second culture medium) is discharged as waste liquid. The cultured cells are recovered by the recovery unit. Summary of the Invention

[0003] For cell culture, it is important to supply nutrients (glucose, glutamine, various amino acids, etc.), gases (oxygen, carbon dioxide, etc.), and waste products (lactic acid, ammonia, etc.). Furthermore, it is also important to supply proteins for cell culture. For this reason, bovine serum (containing albumin and growth factors), growth factors, cytokines, etc. are added to the culture medium.

[0004] If there is insufficient protein supplied to cells, the cells will not grow. On the other hand, if too much protein is supplied to cells, cell growth will be inhibited. Furthermore, the unit price of some proteins (growth factors, cytokines, etc.) is high. For these reasons, it is desirable to appropriately control the amount of protein supplied to cells.

[0005] The present invention aims to solve the above-mentioned problems.

[0006] The first invention is (1) a simulation device that simulates cell growth in a cell culture device, comprising: an acquisition unit that acquires a disappearance rate at which a specified protein in a culture medium is disappeared regardless of the cells, a consumption rate at which the cells consume the protein under first culture conditions, and condition data indicating a second culture condition different from the first culture condition; a simulation execution unit that uses the disappearance rate, the consumption rate, and the condition data acquired by the acquisition unit to simulate a change in concentration of the protein accompanying cell growth under the second culture conditions; and a display unit that acquires the concentration of the protein under the second culture conditions as a result of the simulation, and displays whether the concentration of the protein falls within a predetermined range.

[0007] According to the first invention, a user can acquire culture conditions that can produce high quality cells while suppressing the amount of protein used, by specifying an optimal range of the amount of protein used as the predetermined range.

[0008] (2) In the above-mentioned (1) device, the cell culture device has a bioreactor, a hollow fiber membrane provided inside the bioreactor, the culture region located in an inner hole of the hollow fiber membrane, a non-culture region located inside the bioreactor and outside the hollow fiber membrane, a first supply unit that supplies each of a culture medium containing the protein and cells to the culture region, and a second supply unit that supplies a basal medium not containing the protein to the non-culture region, and it is preferable that the disappearance rate includes a deterioration rate of the protein and a dissolution rate of the protein from the culture region to the non-culture region.

[0009] (3) It is preferable that the device according to (1) or (2) above further comprises an input unit for inputting the disappearance rate and the consumption rate.

[0010] (4) It is preferable that the device of (1) or (2) above further comprises an input unit for inputting the disappearance rate and the change in concentration of the protein measured in the cell culture performed under the first culture conditions, and a calculation unit for calculating the consumption rate based on the concentration of the protein input by the input unit.

[0011] A second invention is (5) a simulation system for simulating cell growth in a cell culture device, comprising: an acquisition unit that acquires a disappearance rate at which a specified protein in a culture medium is disappeared regardless of the cells, a consumption rate at which the cells consume the protein under first culture conditions, and condition data indicating a second culture condition different from the first culture condition; a simulation execution unit that uses the disappearance rate, the consumption rate, and the condition data acquired by the acquisition unit to simulate a change in concentration of the protein accompanying cell growth under the second culture conditions; and a display unit that acquires the concentration of the protein under the second culture conditions as a result of the simulation, and displays whether the concentration of the protein falls within a specified range.

[0012] (6) It is preferable that the system of (5) above comprises a terminal device and a server capable of communicating with each other via a communication network, the terminal device having the display unit, and the server having the acquisition unit and the simulation execution unit.

[0013] (7) A third invention is a simulation method for simulating cell growth in a cell culture device, comprising: an acquisition step of acquiring a disappearance rate at which a specified protein in a culture medium disappears regardless of the cells, a consumption rate at which the cells consume the protein under first culture conditions, and condition data indicating a second culture condition different from the first culture condition; a simulation execution step of simulating a change in concentration of the protein accompanying cell growth under the second culture condition using the disappearance rate, the consumption rate, and the condition data acquired in the acquisition step; and a display step of acquiring the concentration of the protein under the second culture condition as a result of the simulation, and displaying whether the concentration of the protein falls within a specified range.

[0014] According to the present invention, it is possible to estimate the optimal culture conditions. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 is a diagram showing the configuration of a cell culture system. [Diagram 2] FIG. 2 is a diagram showing the configuration of the control unit of the cell culture device. [Diagram 3] FIG. 3 is a diagram illustrating a configuration of the simulation device. [Figure 4] FIG. 4 is a diagram showing an input screen displayed on the display unit. [Diagram 5] FIG. 5 is a diagram showing an input screen displayed on the display unit. [Figure 6] FIG. 6 is a diagram showing a growth data screen displayed on the display unit. [Figure 7] FIG. 7 is a diagram showing a feedback condition screen displayed on the display unit. [Figure 8] FIG. 8 is a diagram showing a result screen displayed on the display unit. [Figure 9] FIG. 9 is a diagram showing a result screen displayed on the display unit. [Figure 10] FIG. 10 is a flow chart showing the flow of a cell culture method carried out using the cell culture system. [Figure 11] FIG. 11 is a flow chart showing the flow of cell culture performed using the cell culture device. [Figure 12] FIG. 12 is a diagram showing the operation of the cell culture device during cell culture. [Figure 13] FIG. 13 is a diagram showing the operation of the cell culture device during cell detachment. [Figure 14] FIG. 14 is a diagram showing the operation of the cell culture device during cell recovery. [Figure 15] FIG. 15 is a diagram showing the configuration of another embodiment of a simulation device. [Figure 16] FIG. 16 is a diagram showing a configuration of a simulation system. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] [1 Configuration of cell culture system 10] FIG. 1 is a diagram showing the configuration of a cell culture system 10. The cell culture system 10 cultures (grows) cells separated from biological tissue in a medium. The cells used in the cell culture system 10 are adherent cells. However, the cells used in the cell culture system 10 may be suspension cells. Specifically, examples of the cells used in the cell culture system 10 include ES cells, iPS cells, mesenchymal stem cells, and the like. The cells used in the cell culture system 10 are not limited to those mentioned above.

[0017] The cell culture system 10 includes a cell culture device 12 and a simulation device 14. The cell culture device 12 includes a cell culture circuit 16, a support device 18, and a control unit 20. A liquid flows in the cell culture circuit 16. The liquid includes at least one of a cell liquid, a culture medium, a cleaning liquid, and a peeling liquid. The cell liquid is a solution containing cells. The culture medium is a culture liquid for growing cells. The culture medium is selected according to the cells to be cultured. In this embodiment, two types of media are used, a basal medium and a culture medium. For example, MEM (Minimum Essential Media) is used as the basal medium. For example, MEM is used as the culture medium. Albumin, growth factors, cytokines, etc. are used as proteins. For example, bovine serum containing albumin, growth factors, etc. is added to the basal medium. The cleaning liquid cleans the inside of the cell culture circuit 16. For example, water, a buffer solution, physiological saline, etc. are used as the cleaning liquid. Examples of the buffer solution include PBS (Phosphate Buffered Saline) and TBS (Tris-Buffered Saline). The detachment solution detaches cells from a bioreactor 30 (described later) of the cell culture circuit 16. For example, trypsin or EDTA solution is used. The medium, washing solution, and detachment solution are not limited to the above-mentioned liquids.

[0018] [1-1 Cell culture device 12] [1-1-1 Cell culture circuit 16] The cell culture circuit 16 is discarded after a single use. In other words, the cell culture circuit 16 is discarded every time a predetermined number of cells are cultured. In other words, the cell culture circuit 16 is a disposable item. The cell culture circuit 16 includes a supply unit 22, a collection container 24, a waste liquid storage unit 26, and a culture body 28.

[0019] The supply unit 22 supplies liquids such as cell liquid, culture medium, washing liquid, and peeling liquid to the culture body 28. The supply unit 22 has a first supply unit 22a and a second supply unit 22b. The collection container 24 collects cells cultured in the culture body 28. The waste liquid storage unit 26 stores waste liquid generated in the culture body 28. Each of the collection container 24 and the waste liquid storage unit 26 is, for example, a medical bag made of a soft resin material in a bag shape. Each of the collection container 24 and the waste liquid storage unit 26 may be a tank or the like made of a hard material.

[0020] The culture body 28 has a bioreactor 30, a flow path 32, a gas exchange section 34, a first sampling section 35, a sensor section 36, and a second sampling section 38.

[0021] The bioreactor 30 has a plurality of hollow fiber membranes 40 and a cylindrical housing 42. The plurality of hollow fiber membranes 40 are accommodated in the housing 42. One end of each hollow fiber membrane 40 is fixed to one end of the housing 42. The other end of each hollow fiber membrane 40 is fixed to the other end of the housing 42. Each hollow fiber membrane 40 is made of, for example, a polymer material.

[0022] The bioreactor 30 includes a first region 44 and a second region 46. The first region 44 is an inner hole of the plurality of hollow fiber membranes 40. The first region 44 is a region in the culture medium where cells are present. This region is also called a culture region. The second region 46 is a space between the inner peripheral surface of the housing 42 and the outer peripheral surfaces of the plurality of hollow fiber membranes 40. The second region 46 is a non-culture region in the inner region of the bioreactor 30. Each hollow fiber membrane 40 has a plurality of pores (not shown). The first region 44 and the second region 46 are connected to each other through the plurality of pores of each hollow fiber membrane 40. The diameter of each pore is a size that prevents the passage of macromolecules (cells, etc.) while allowing the passage of small molecules (e.g., water, ions, oxygen, lactate, etc.). The diameter of each pore is set, for example, to be 0.005 [μm] or more and 10 [μm] or less.

[0023] A first inlet port 48, a first outlet port 50, a second inlet port 52, and a second outlet port 54 are attached to the housing 42. The first inlet port 48 is attached to one end of the housing 42. The first inlet port 48 communicates with the first region 44 via an inlet located at one end of the plurality of hollow fiber membranes 40. The first outlet port 50 is attached to the other end of the housing 42. The first outlet port 50 communicates with the first region 44 via an outlet located at the other end of the plurality of hollow fiber membranes 40.

[0024] The second inlet port 52 and the second outlet port 54 are attached to the outer circumferential surface of the housing 42. The second inlet port 52 is located between the center of the housing 42 and the first inlet port 48 in the longitudinal direction of the housing 42. The second outlet port 54 is located between the center of the housing 42 and the first outlet port 50 in the longitudinal direction of the housing 42. The second inlet port 52 and the second outlet port 54 each communicate with the second region 46.

[0025] The flow path 32 includes a plurality of tubes through which the liquid flows. Each tube is made of a soft resin material. The flow path 32 includes a first supply flow path 56, a first circulation flow path 58, a second supply flow path 60, a second circulation flow path 62, a recovery flow path 64, and a waste liquid flow path 66.

[0026] One end of the first supply flow path 56 is connected to the first supply unit 22a. The first supply unit 22a supplies the cell liquid, the culture medium, the washing solution, and the stripping solution one by one at a predetermined timing to the first supply flow path 56. The other end of the first supply flow path 56 is connected to a first junction 68 of the first circulation flow path 58.

[0027] The first junction 68 is located in the middle of the extension direction of the first circulation flow path 58. One end of the first circulation flow path 58 is connected to the first inlet port 48. The other end of the first circulation flow path 58 is connected to the first outlet port 50. The first circulation flow path 58 communicates with the inner holes (first regions 44) of the multiple hollow fiber membranes 40.

[0028] One end of the second supply flow path 60 is connected to the second supply unit 22b. The second supply unit 22b supplies the basal medium and the washing solution one by one at a predetermined timing to the second supply flow path 60. The other end of the second supply flow path 60 is connected to a second junction 70 of the second circulation flow path 62.

[0029] The second junction 70 is located at the middle of the second circulation flow path 62 in the extending direction. One end of the second circulation flow path 62 is connected to the second inlet port 52. The other end of the second circulation flow path 62 is connected to the second outlet port 54. The second circulation flow path 62 communicates with the space (second region 46) between the multiple hollow fiber membranes 40 and the housing 42. Hereinafter, the first circulation flow path 58 and the second circulation flow path 62 may be collectively referred to as the "circulation flow path 72."

[0030] The recovery flow path 64 extends from the first circulation flow path 58. One end of the recovery flow path 64 is connected to a recovery branch portion 74 of the first circulation flow path 58. The recovery branch portion 74 is located between the first junction portion 68 and the first outlet port 50 in the first circulation flow path 58. The other end of the recovery flow path 64 is connected to the recovery container 24.

[0031] The waste liquid flow path 66 flows the liquid to be discarded from the circulation flow path 72. The waste liquid flow path 66 has a first waste liquid flow path 76, a second waste liquid flow path 78, and a third waste liquid flow path 80. The first waste liquid flow path 76 extends from the first circulation flow path 58. One end of the first waste liquid flow path 76 is connected to a first branching portion 82 of the first circulation flow path 58. The first branching portion 82 is located between the first outlet port 50 and the recovery branching portion 74 of the first circulation flow path 58. The second waste liquid flow path 78 extends from the second circulation flow path 62. One end of the second waste liquid flow path 78 is connected to a second branching portion 84 of the second circulation flow path 62. The second branching portion 84 is located between the second junction portion 70 and the second outlet port 54 of the second circulation flow path 62. The other end of the first waste liquid flow path 76 and the other end of the second waste liquid flow path 78 are connected to each other at an intermediate junction 86. One end of the third waste liquid flow path 80 is connected to the first waste liquid flow path 76 and the second waste liquid flow path 78 at the intermediate junction 86. The other end of the third waste liquid flow path 80 is connected to the waste liquid storage section 26.

[0032] The gas exchange unit 34 is attached to the second circulation flow path 62 between the second junction 70 and the second inlet port 52. The gas exchange unit 34 passes a gas of a predetermined composition through the liquid (basal medium) flowing through the second circulation flow path 62. The gas used in the gas exchange unit 34 has, for example, compositions similar to those of air in nature. In other words, the gas contains nitrogen, oxygen, and carbon dioxide. Specifically, the gas contains, for example, 75% nitrogen, 20% oxygen, and 5% carbon dioxide by volume.

[0033] The first sampling unit 35 is connected to the first circulation flow path 58. The first sampling unit 35 extracts a portion of the liquid (culture medium) flowing through the first circulation flow path 58. The first sampling unit 35 aseptically collects a tube fragment containing the internal liquid from a tube of sufficient length using, for example, a sterile joining device.

[0034] The sensor unit 36 ​​is attached to the third waste liquid flow path 80. The sensor unit 36 ​​has a gas sensor 88 and a pH sensor 90. The gas sensor 88 measures the gas concentration of the liquid flowing through the third waste liquid flow path 80. Specifically, the gas sensor 88 includes an oxygen sensor and a carbon dioxide sensor. The oxygen sensor measures the oxygen concentration of the liquid flowing through the third waste liquid flow path 80. The carbon dioxide sensor measures the carbon dioxide concentration of the liquid flowing through the third waste liquid flow path 80. The pH sensor 90 measures the pH (hydrogen ion exponent) of the liquid flowing through the third waste liquid flow path 80. Each of the gas sensor 88 and the pH sensor 90 outputs the measurement results to the control unit 20.

[0035] The second sampling section 38 is connected to a portion of the third waste liquid flow path 80 between the sensor section 36 and the waste liquid storage section 26. The second sampling section 38 extracts a portion of the liquid flowing through the third waste liquid flow path 80 and measures components contained in the liquid. The second sampling section 38 has a biosensor 92, a flow path (not shown), and the like.

[0036] The biosensor 92 has a glucose sensor 94 and a lactate sensor 96. The glucose sensor 94 measures the glucose concentration of the liquid extracted from the third waste liquid flow path 80. The lactate sensor 96 measures the lactate concentration of the liquid extracted from the third waste liquid flow path 80. Each of the glucose sensor 94 and the lactate sensor 96 outputs the measurement results to the control unit 20.

[0037] [1-1-2 Support device 18] The above-mentioned cell culture circuit 16 is set in a support device 18. The support device 18 has a cassette that supports the cell culture circuit 16. The support device 18 is a reusable item that can be used multiple times.

[0038] The support device 18 includes a plurality of pumps 98 and a plurality of clamps 100. Each of the plurality of pumps 98 applies a flow force to the liquid in the flow path 32 by squeezing the wall of the flow path 32. Each of the plurality of pumps 98 has a pressing member (not shown). The pressing member includes, for example, a rotating member and a plurality of pressing rollers. The plurality of pressing rollers are attached to the outer periphery of the rotating member. The plurality of pressing rollers are arranged at intervals in the circumferential direction of the rotating member. Each pressing roller rubs the outer surface of the wall of the flow path 32.

[0039] The multiple pumps 98 include a first supply pump 102, a first circulation pump 104, a second supply pump 106, and a second circulation pump 108. Note that, as shown in Fig. 1, a state in which the cell culture circuit 16 is set on the support device 18 is simply referred to as a "set state."

[0040] In the set state, a part of the first supply flow path 56 is attached to the first supply pump 102. The first supply pump 102 applies a flow force to the liquid in the first supply flow path 56 in a direction from the supply unit 22 to the first circulation flow path 58.

[0041] In the set state, a part of the first circulation flow path 58 is attached to the first circulation pump 104. The first circulation pump 104 imparts a flow force to the liquid in the first circulation flow path 58 in a direction from the first outlet port 50 to the first inlet port 48. Note that the first circulation pump 104 can also impart a flow force to the liquid in the first circulation flow path 58 in a direction from the first inlet port 48 to the first outlet port 50.

[0042] In the set state, a part of the second supply flow path 60 is attached to the second supply pump 106. The second supply pump 106 applies a flow force to the liquid in the second supply flow path 60 in a direction from the supply unit 22 to the second circulation flow path 62.

[0043] In the set state, a part of the second circulation flow path 62 is attached to the second circulation pump 108. The second circulation pump 108 imparts a flow force to the liquid in the second circulation flow path 62 in a direction from the second outlet port 54 to the second inlet port 52. Note that the second circulation pump 108 can also impart a flow force to the liquid in the second circulation flow path 62 in a direction from the second inlet port 52 to the second outlet port 54.

[0044] The clamps 100 press the outer surface of the flow path 32 toward the inner surface thereof to close the flow path 32. For example, the clamps 100 are on-off valves. The clamps 100 include a recovery clamp 110, a first waste liquid clamp 112, a second waste liquid clamp 114, and a third waste liquid clamp 116.

[0045] In the set state, a portion of the recovery flow path 64 is attached to the recovery clamp 110. The recovery clamp 110 opens and closes the recovery flow path 64. In the set state, a portion of the first waste liquid flow path 76 is attached to the first waste liquid clamp 112. The first waste liquid clamp 112 opens and closes the first waste liquid flow path 76. In the set state, a portion of the second waste liquid flow path 78 is attached to the second waste liquid clamp 114. The second waste liquid clamp 114 opens and closes the second waste liquid flow path 78. In the set state, a portion of the third waste liquid flow path 80 is attached to the third waste liquid clamp 116. The third waste liquid clamp 116 opens and closes the third waste liquid flow path 80.

[0046] [1-1-3 Control unit 20] 2 is a diagram showing the configuration of the control unit 20 of the cell culture device 12. The control unit 20 has a first calculation unit 118, a first storage unit 120, and various driving circuits (not shown).

[0047] The first calculation unit 118 has a processing circuit. The processing circuit may be a processor such as a CPU. The processing circuit may be an integrated circuit such as an ASIC or an FPGA. The processor can execute various processes by executing a program stored in the first storage unit 120. The control unit 20 functions as a pump control unit 122, a clamp control unit 124, a gas exchange control unit 126, and a measurement unit 128. At least a part of the processes may be executed by an electronic circuit including a discrete device.

[0048] The pump control unit 122 controls each of the multiple pumps 98. Specifically, the pump control unit 122 outputs a command signal to a pump drive circuit (not shown). The pump drive circuit supplies each of the multiple pumps 98 with power in accordance with the command signal from the pump control unit 122. The clamp control unit 124 controls each of the multiple clamps 100. Specifically, the clamp control unit 124 outputs a command signal to a clamp drive circuit (not shown). The clamp drive circuit supplies each of the multiple clamps 100 with power in accordance with the command signal from the clamp control unit 124. The gas exchange control unit 126 controls the gas exchange unit 34. Specifically, the gas exchange control unit 126 outputs a command signal to a gas exchanger drive circuit (not shown). The gas exchanger drive circuit supplies the gas exchange unit 34 with power in accordance with the command signal from the gas exchange control unit 126. The measurement unit 128 acquires measurement results from each of the gas sensor 88, the pH sensor 90, the glucose sensor 94, and the lactate sensor 96. The measurement section 128 stores the acquired measurement results in the first storage section 120.

[0049] The first storage unit 120 has a volatile memory and a non-volatile memory. An example of the volatile memory is a RAM. The volatile memory is used as a working memory of the processor. The volatile memory temporarily stores data required for processing or calculation. An example of the non-volatile memory is a ROM, a flash memory, etc. The non-volatile memory is used as a storage memory. The non-volatile memory stores programs, tables, maps, etc. At least a part of the first storage unit 120 may be provided in the processor, integrated circuit, etc. as described above.

[0050] [1-2 Simulation device 14] 3 is a diagram showing a configuration of the simulation device 14. The simulation device 14 has an input unit 130, a simulation unit 132, and a display unit 134. As the simulation device 14, a personal computer, a smartphone, a tablet, or the like may be used.

[0051] The input unit 130 has a human-machine interface such as a keyboard, a mouse, a touch pad, etc. The input unit 130 may also have a human-machine interface integrated with the display unit 134, such as a touch panel. The input unit 130 can input data corresponding to operations performed by the user to the simulation unit 132.

[0052] The simulation unit 132 has a second calculation unit 136 and a second storage unit 138. The first calculation unit 118 and the first storage unit 120 may be used as the second calculation unit 136 and the second storage unit 138. In other words, the control unit 20 of the cell culture device 12 may be used as the simulation unit 132. The second calculation unit 136 has a processing circuit. The processing circuit may be a processor such as a CPU. The processing circuit may be an integrated circuit such as an ASIC or an FPGA. The processor can execute various processes by executing a program stored in the second storage unit 138. The simulation unit 132 functions as an acquisition unit 140, a simulation execution unit 142, and a display control unit 144. At least a part of the multiple processes may be executed by an electronic circuit including a discrete device.

[0053] The acquisition unit 140 acquires data from outside the second calculation unit 136. For example, the acquisition unit 140 can acquire data from the input unit 130. Also, the acquisition unit 140 can acquire data designated by the input unit 130 from the second storage unit 138. Also, the acquisition unit 140 can acquire data designated by the input unit 130 from a device (such as the control unit 20) designated by the input unit 130. The simulation execution unit 142 uses the data acquired by the acquisition unit 140 to simulate cell growth and protein concentration changes by the cell culture device 12. The display control unit 144 causes the display unit 134 to display various screens. For example, the display control unit 144 can cause the display unit 134 to display the data stored in the second storage unit 138. The display control unit 144 can cause the display unit 134 to display the results of the simulation executed by the simulation execution unit 142.

[0054] The second storage unit 138 has a volatile memory and a non-volatile memory. Examples of the volatile memory include RAM. The volatile memory is used as a working memory of the processor. The volatile memory temporarily stores data required for processing or calculation. Examples of the non-volatile memory include ROM and flash memory. The non-volatile memory is used as a storage memory. The non-volatile memory stores programs, tables, maps, and the like. In this embodiment, the non-volatile memory stores a simulation program executed by the simulation execution unit 142. Furthermore, the non-volatile memory stores defaults for various data related to cell proliferation. At least a part of the second storage unit 138 may be provided in the processor, integrated circuit, and the like described above.

[0055] The display unit 134 has a human-machine interface such as a display. The display unit 134 may have a human-machine interface integrated with the input unit 130, such as a touch panel. The display unit 134 can display various screens described in [2] below.

[0056] [2 screens] The display unit 134 can display an input screen 146 (FIGS. 4 and 5), a growth data screen 148 (FIG. 6), a feedback condition screen 150 (FIG. 7), a result screen 152 (FIGS. 8 and 9), and the like.

[0057] [2-1 Input screen 146] 4 and 5 are diagrams showing an input screen 146 displayed on the display unit 134. FIG. 4 shows an upper part of the input screen 146, and FIG. 5 shows a lower part of the input screen 146. When the user performs a scroll operation downward with the input screen 146 of FIG. 4 displayed on the display unit 134, the input screen 146 of FIG. 5 is displayed on the display unit 134. The input screen 146 is a screen for inputting various data used in the simulation of cell culture. When the user operates the input unit 130, the display unit 134 displays the input screen 146.

[0058] The input screen 146 has a scale field 154 (FIG. 4). The scale field 154 is an input field for specifying the scale of cell culture in the simulation. The user can select a scale from a drop-down list displayed in the scale field 154.

[0059] The input screen 146 has a cell type field 156 (FIG. 4). The cell type field 156 is an input field for specifying proliferation data to be used in the simulation. The proliferation data is data showing the proliferation state of cells under any culture conditions. The proliferation data specified in the cell type field 156 is a proliferation model of cells for the simulation. The proliferation data is created based on data actually measured in a cell culture process carried out in the past. The second storage unit 138 stores default proliferation data. In addition, the second storage unit 138 can store data actually measured in the cell culture process shown in step S5 of FIG. 10 as proliferation data. A specific example of the proliferation data is shown in FIG. 6. The user can select either the default or the actual measurement result from a drop-down list displayed in the cell type field 156.

[0060] The input screen 146 has a feedback field 160 (FIG. 4). The feedback field 160 is an input field for specifying whether or not feedback conditions are to be used in the simulation. A specific example of the feedback conditions is shown in FIG. 7. The user can select either "on" or "off" from a drop-down list displayed in the feedback field 160. When "on" is selected, the feedback conditions are used in the simulation. When "off" is selected, the feedback conditions are not used in the simulation.

[0061] The input screen 146 has a culture medium input field 162 (FIG. 4). The culture medium input field 162 is an input field for specifying data of the culture medium to be circulated through the first circulation flow path 58 in the simulation. The culture medium data includes the glucose concentration, the lactic acid concentration, the concentration of one or more proteins, the type of culture medium, pka, the unit price of the culture medium, and the like. There may be one type of protein or multiple types of proteins. The user can specify the protein to be added to the culture medium in the culture medium input field 162. The culture medium data is condition data that indicates the culture conditions of the simulation.

[0062] The input screen 146 has a basal medium input field 164 (FIG. 4). The basal medium input field 164 is an input field for specifying data of the basal medium to be circulated through the second circulation flow path 62 in the simulation. The data of the basal medium includes the glucose concentration, the lactic acid concentration, the type of basal medium, pka, the unit price of the basal medium, etc. The data of the basal medium is condition data indicating the culture conditions of the simulation.

[0063] The input screen 146 has a gas input field 166 (FIG. 4). The gas input field 166 is an input field for designating data of gas used in the gas exchange unit 34 in the simulation. The gas data includes the volume ratio of oxygen contained in the gas, the volume ratio of carbon dioxide contained in the gas, the gas flow rate, etc. The gas data is condition data indicating the culture conditions of the simulation.

[0064] The input screen 146 has an other input field 168 (FIG. 4). The other input field 168 is an input field for specifying other data related to the culture medium. The other data includes the capacity of the first circulation flow path 58, the capacity of the second circulation flow path 62, the atmospheric pressure, the water vapor pressure, etc. The other data is condition data indicating the culture conditions of the simulation.

[0065] The input screen 146 has a pump speed input field 170 (FIG. 4). The pump speed input field 170 is an input field for specifying the flow rate of each pump 98 in the simulation. The flow rate of each pump 98 is set for each day of the culture period. The flow rate of each pump 98 is condition data indicating the culture conditions of the simulation.

[0066] The input screen 146 has a number of days input field 172 (FIG. 4). The number of days input field 172 is an input field for specifying the number of days for which cells are cultured in the simulation. The number of days for which cells are cultured is condition data that indicates the culture conditions in the simulation.

[0067] The input screen 146 has a seeding number input field 174 (FIG. 4). The seeding number input field 174 is an input field for specifying the seeding number in the simulation. The seeding number is condition data indicating the culture conditions of the simulation.

[0068] The input screen 146 has a doubling time input field 176 (FIG. 4). The doubling time input field 176 is an input field for specifying the time it takes for cells to double (doubling time) in the simulation. The doubling time is condition data that indicates the culture conditions of the simulation.

[0069] The input screen 146 has a temperature input field 178 (FIG. 4). The temperature input field 178 is an input field for specifying an environmental temperature in the simulation. The environmental temperature is condition data indicating the culture conditions in the simulation.

[0070] The input screen 146 has a threshold input field 180 (FIG. 4). The threshold input field 180 is an input field for specifying the respective thresholds of the glucose concentration, the lactate concentration, the partial pressure of oxygen, the partial pressure of carbon dioxide, pH, and protein. As the threshold, at least one of the lower limit value (LLR), the lower warning value (LAR), the upper limit value (ULR), and the upper warning value (UAR) is specified. For example, only the lower limit value (LLR) and the lower warning value (LAR) may be specified. For example, only the upper limit value (ULR) and the upper warning value (UAR) may be specified. For example, the lower limit value (LLR), the lower warning value (LAR), the upper limit value (ULR), and the upper warning value (UAR) may be specified. The user can arbitrarily specify the threshold.

[0071] The input screen 146 has a protein parameter field 182 (FIG. 5). The protein parameter field 182 is an input field for specifying the rate at which various proteins disappear in the simulation for each disappearance factor. There are three main factors for protein disappearance. The first disappearance factor is protein degradation over time. The protein parameter field 182 has an input field for specifying the protein degradation rate. The second disappearance factor is protein elution. Inside the bioreactor 30, the components of the culture medium in the culture region pass through each pore of the hollow fiber membrane 40 and are eluted into the non-culture region. That is, a part of the protein contained in the culture medium is eluted from the culture region to the non-culture region. The protein eluted into the non-culture region does not contribute to cell culture. The protein parameter field 182 has an input field for specifying the protein elution rate. The third disappearance factor is protein consumption by cells. The protein parameter field 182 has an input field for specifying the protein consumption rate by cells. The deterioration rate and elution rate are parameters that are determined regardless of the presence or absence of cells. The dissolution rate is a parameter determined by the cell culture device 12. Note that the user can specify the protein to be used in the cell culture and the disappearance rates of the three proteins in the protein parameter field 182.

[0072] Save button 183 (FIG. 5) is a button for saving data specified in each input field. The user can press save button 183 by operating input unit 130. When save button 183 is pressed, second storage unit 138 stores the data specified in each input field.

[0073] [2-2 Breeding Data Screen 148] 6 is a diagram showing a growth data screen 148 displayed on the display unit 134. The growth data screen 148 is a screen showing each growth data. When the user operates the input unit 130, the display unit 134 displays the growth data screen 148. The second storage unit 138 stores each growth data as a data set.

[0074] The proliferation data screen 148 has a biodata graph 184. In the biodata graph 184, the horizontal axis represents time, and the vertical axis represents the metabolic rate of the biodata. The biodata graph 184 displays a metabolic rate line 186 and a metabolic rate line 188. The metabolic rate line 186 shows the progress of the metabolic rate of glucose. The metabolic rate line 188 shows the progress of the metabolic rate of lactate. The metabolic rate of the biodata is proliferation data that indicates the proliferation state of cells.

[0075] The proliferation data screen 148 has a gas data graph 190. In the gas data graph 190, the horizontal axis represents time, and the vertical axis represents the metabolic rate of the biodata. The gas data graph 190 displays a metabolic rate line 192 and a metabolic rate line 194. The metabolic rate line 192 shows the progress of the metabolic rate of oxygen. The metabolic rate line 194 shows the progress of the metabolic rate of carbon dioxide. The metabolic rate of the gas data is proliferation data that indicates the proliferation state of the cells.

[0076] The proliferation data screen 148 has a cell graph 196. In the cell graph 196, the horizontal axis represents time and the vertical axis represents the number of cells. A cell number line 198 is displayed on the cell graph 196. The cell number line 198 indicates the progress of the number of cells. The cell number is proliferation data that indicates the proliferation state of the cells.

[0077] [2-3 Feedback Condition Screen 150] 7 is a diagram showing a feedback condition screen 150 displayed on display unit 134. Feedback condition screen 150 is a screen for inputting feedback conditions and change data. Feedback conditions are conditions for changing condition data according to the state of a simulation during a simulation. Change data is a change value of the condition data. When a user operates input unit 130, display unit 134 displays feedback condition screen 150.

[0078] The feedback condition screen 150 has a condition field 200 and a data field 202. The condition field 200 is an input field for specifying the feedback condition. The data field 202 is an input field for specifying the change data. For example, the condition field 200 and the data field 202 shown in No. 1 of FIG. 7 mean that "when lactic acid becomes greater than XXX [mM], the flow rate of the first circulation pump 104 is set to XXX [mL / min]." The feedback condition and the change data are condition data indicating the culture conditions of the simulation. Although not shown, feedback conditions such as glucose, carbon dioxide, pH, and various proteins can also be specified.

[0079] When the save button 183 on the input screen 146 is pressed, the second storage unit 138 stores the data designated in each input field on the feedback condition screen 150.

[0080] [2-4 Result screen 152] 8 and 9 are diagrams showing result screen 152 displayed on display unit 134. FIG. 8 shows the upper part of result screen 152, and FIG. 9 shows the lower part of result screen 152. When the user performs a downward scroll operation while result screen 152 of FIG. 8 is displayed on display unit 134, result screen 152 of FIG. 9 is displayed on display unit 134. Result screen 152 is a screen showing the results of the simulation performed in step S2 of FIG. 10. After the simulation, the user operates input unit 130, causing display unit 134 to display result screen 152.

[0081] The result screen 152 has a waste amount column 204 and a cost column 206. The waste amount column 204 displays the total amount of medium wasted in the simulated cell culture. The cost column 206 displays the cost in the simulated cell culture.

[0082] The result screen 152 has a glucose graph 208 (FIG. 8). In the glucose graph 208, the horizontal axis represents time, and the vertical axis represents the glucose concentration. The glucose graph 208 displays a concentration line 210, a warning line 212, and a lower limit line 214. The concentration line 210 indicates the transition of the glucose concentration during the culture period. The warning line 212 indicates the boundary value between the OK range and the warning range. The lower limit line 214 indicates the boundary value between the warning range and the NG range. The boundary value indicated by the warning line 212 is the lower warning value of the glucose concentration inputted in the threshold input field 180 of the input screen 146. The boundary value indicated by the lower limit line 214 is the lower limit value of the glucose concentration inputted in the threshold input field 180 of the input screen 146. The range above the warning line 212 is the OK range. The range below the lower limit line 214 is the NG range. The range between the warning line 212 and the lower limit line 214 is the warning range. It is preferable that the concentration line 210 is in the OK range above the warning line 212. That is, it is preferable that the glucose concentration is within the OK range at all times during the culture period.

[0083] The result screen 152 has a lactate graph 216 (FIG. 8). In the lactate graph 216, the horizontal axis represents time, and the vertical axis represents the lactate concentration. The lactate graph 216 displays a concentration line 218, a warning line 220, and an upper limit line 222. The concentration line 218 indicates the change in lactate concentration during the culture period. The warning line 220 indicates the boundary value between the OK range and the warning range. The upper limit line 222 indicates the boundary value between the warning range and the NG range. The boundary value indicated by the warning line 220 is the upper warning value of the lactate concentration input in the threshold input field 180 of the input screen 146. The boundary value indicated by the upper limit line 222 is the upper limit value of the lactate concentration input in the threshold input field 180 of the input screen 146. The range below the warning line 220 is the OK range. The range above the upper limit line 222 is the NG range. The range between warning line 220 and upper limit line 222 is the warning range. Concentration line 218 is preferably in the OK range below warning line 220. That is, the lactate concentration is preferably in the OK range at all times during the culture period.

[0084] The result screen 152 has an O2 graph 224 (FIG. 8). In the O2 graph 224, the horizontal axis represents time, and the vertical axis represents the partial pressure of oxygen. The O2 graph 224 displays a partial pressure line 226, a warning line 228, and a lower limit line 230. The partial pressure line 226 indicates the transition of the partial pressure of oxygen during the culture period. The warning line 228 indicates the boundary value between the OK range and the warning range. The lower limit line 230 indicates the boundary value between the warning range and the NG range. The boundary value indicated by the warning line 228 is the lower warning value of the partial pressure of oxygen inputted in the threshold input field 180 of the input screen 146. The boundary value indicated by the lower limit line 230 is the lower limit value of the partial pressure of oxygen inputted in the threshold input field 180 of the input screen 146. The range above the warning line 228 is the OK range. The range below the lower limit line 230 is the NG range. The range between warning line 228 and lower limit line 230 is the warning range. Partial pressure line 226 is preferably in the OK range above warning line 228. That is, the partial pressure of oxygen is preferably in the OK range at all times during the culture period.

[0085] The result screen 152 has a CO2 graph 232 (FIG. 8). In the CO2 graph 232, the horizontal axis represents time, and the vertical axis represents the partial pressure of carbon dioxide. The CO2 graph 232 displays a partial pressure line 234, a warning line 236, and an upper limit line 238. The partial pressure line 234 indicates the transition of the partial pressure of carbon dioxide during the culture period. The warning line 236 indicates the boundary value between the OK range and the warning range. The upper limit line 238 indicates the boundary value between the warning range and the NG range. The boundary value indicated by the warning line 236 is the upper warning value of the partial pressure of carbon dioxide inputted in the threshold input field 180 of the input screen 146. The boundary value indicated by the upper limit line 238 is the upper limit value of the partial pressure of carbon dioxide inputted in the threshold input field 180 of the input screen 146. The range below the warning line 236 is the OK range. The range above the upper limit line 238 is the NG range. The range between warning line 236 and upper limit line 238 is the warning range. Partial pressure line 234 is preferably in the OK range below warning line 236. That is, the partial pressure of carbon dioxide is preferably in the OK range at all times during the culture period.

[0086] The result screen 152 has a pH graph 240 (FIG. 8). In the pH graph 240, the horizontal axis represents time, and the vertical axis represents the pH of the medium. The pH graph 240 displays a pH line 242, a lower warning line 244, a lower limit line 246, an upper warning line 248, and an upper limit line 250. The pH line 242 indicates the transition of pH during the culture period. The lower warning line 244 indicates the boundary value between the OK range and the lower warning range. The lower limit line 246 indicates the boundary value between the lower warning range and the first NG range. The upper warning line 248 indicates the boundary value between the OK range and the upper warning range. The upper limit line 250 indicates the boundary value between the upper warning range and the second NG range. The boundary value indicated by the lower warning line 244 is the lower warning value of pH inputted in the threshold input field 180 of the input screen 146. The boundary value indicated by the lower limit line 246 is the lower limit value of pH inputted in the threshold input field 180 of the input screen 146. The boundary value indicated by the upper warning line 248 is the upper warning value of pH inputted in the threshold input field 180 of the input screen 146. The boundary value indicated by the upper limit line 250 is the upper limit value of pH inputted in the threshold input field 180 of the input screen 146. The range between the lower warning line 244 and the upper warning line 248 is the OK range. The range below the lower limit line 246 is the first NG range. The range between the lower warning line 244 and the lower limit line 246 is the lower warning range. The range above the upper limit line 250 is the second NG range. The range between the upper warning line 248 and the upper limit line 250 is the upper warning range. It is preferable that the pH line 242 is in the OK range between the lower warning line 244 and the upper warning line 248. That is, the pH of the medium is preferably within the OK range throughout the entire culture period.

[0087] The result screen 152 has a flow rate graph 252 (FIG. 8). In the flow rate graph 252, the horizontal axis represents time, and the vertical axis represents the flow rate of the first circulating pump 104 and the flow rate of the second circulating pump 108. A first flow rate line 254 and a second flow rate line 256 are displayed on the flow rate graph 252. The first flow rate line 254 indicates the transition of the flow rate of the first circulating pump 104 during the culture period. The second flow rate line 256 indicates the transition of the flow rate of the second circulating pump 108 during the culture period.

[0088] The result screen 152 has an albumin graph 258 ( FIG. 9 ). In the albumin graph 258, the horizontal axis represents time, and the vertical axis represents the albumin concentration. The albumin graph 258 displays a concentration line 260, a lower warning line 262, a lower limit line 264, an upper warning line 266, and an upper limit line 268. The concentration line 260 indicates the transition of the albumin concentration during the culture period. The lower warning line 262 indicates the boundary value between the OK range and the lower warning range. The lower limit line 264 indicates the boundary value between the lower warning range and the first NG range. The upper warning line 266 indicates the boundary value between the OK range and the upper warning range. The upper limit line 268 indicates the boundary value between the upper warning range and the second NG range. The boundary value indicated by the lower warning line 262 is the lower warning value of the albumin concentration input in the threshold input field 180 of the input screen 146. The boundary value indicated by the lower limit line 264 is the lower limit value of the albumin concentration input in the threshold input field 180 of the input screen 146. The boundary value indicated by the upper warning line 266 is the upper warning value of the albumin concentration input in the threshold input field 180 of the input screen 146. The boundary value indicated by the upper limit line 268 is the upper limit value of the albumin concentration input in the threshold input field 180 of the input screen 146. The range between the lower warning line 262 and the upper warning line 266 is the OK range. The range below the lower limit line 264 is the first NG range. The range between the lower warning line 262 and the lower limit line 264 is the lower warning range. The range above the upper limit line 268 is the second NG range. The range between the upper warning line 266 and the upper limit line 268 is the upper warning range. Concentration line 260 is preferably in the OK range between lower warning line 262 and upper warning line 266. That is, the albumin concentration is preferably in the OK range at all times during the incubation period.

[0089] The result screen 152 has a bFGF graph 270 (FIG. 9). The bFGF graph 270 shows the transition of the bFGF concentration during the culture period. The bFGF graph 270 can be explained by replacing "albumin" in the explanation of the albumin graph 258 above with "bFGF."

[0090] The result screen 152 can also show the transition of the concentration of other proteins. In other words, the result screen 152 can show the transition of the concentration of the protein specified in the culture medium input field 162 of the input screen 146.

[0091] [3 Cell culture method] Fig. 10 is a flowchart showing the flow of a cell culture method performed using the cell culture system 10. Steps S1 to S3 in Fig. 10 are performed by the simulation device 14. Step S5 in Fig. 10 is performed by the cell culture device 12. Details of step S5 are shown in Fig. 11. Steps S4 and S6 are determined by the user.

[0092] Before performing the simulation, the user measures the disappearance rates of the three proteins described above. For example, the user measures the degradation rate of the protein by any method. The user also measures the elution rate of the protein without supplying cells to the inner hole of the hollow fiber membrane 40. In this case, the user may use the cell culture device 12 or another device equipped with the hollow fiber membrane 40.

[0093] Furthermore, the user actually performs cell culture using the cell culture device 12 in order to measure the protein consumption rate by the cells. The user measures the protein concentration in the culture medium extracted by the first sampling unit 35. The measurement is performed, for example, by the BCA method, the Bardforf method, or the like. The user can obtain the transition of the protein concentration by performing the measurement over time. The user can calculate the protein consumption rate by the cells from the transition of the protein concentration, the protein degradation rate, and the protein elution rate. The process from measuring the protein concentration of the sampled culture medium to calculating the protein consumption rate can also be automated.

[0094] Before step S1, the user operates the input unit 130 to start the simulation program. In response to the user's operation, the second calculation unit 136 executes the simulation program stored in the second storage unit 138. Then, the display unit 134 displays the input screen 146 shown in Figs. 4 and 5.

[0095] In step S1, the user operates the input unit 130 to specify data in each input field of the input screen 146. For example, in the first simulation, the user specifies the default in the cell type field 156. The user also inputs the disappearance rate (deterioration rate, dissolution rate, consumption rate) of each protein in the protein parameter field 182. After specifying each piece of data, the user presses the save button 183. The input unit 130 inputs the data in each input field to the simulation unit 132. The second storage unit 138 stores each piece of data. When step S1 ends, the process proceeds to step S2.

[0096] In step S2, the user operates the input unit 130 to start a simulation of cell culture. In response to an instruction from the input unit 130, the simulation execution unit 142 starts a simulation of cell culture using each data (growth data, condition data, and various parameters of protein) stored in the second storage unit 138. The simulation execution unit 142 simulates cell culture for a specified culture period using the growth data, condition data, and various parameters of protein. The simulation execution unit 142 calculates the amount of each component contained in the medium at each time during the culture period. Specifically, the simulation execution unit 142 calculates the concentration of glucose at each time during the culture period. The simulation execution unit 142 also calculates the concentration of lactic acid at each time during the culture period. The simulation execution unit 142 also calculates the partial pressure of oxygen at each time during the culture period. The simulation execution unit 142 also calculates the partial pressure of carbon dioxide at each time during the culture period. The simulation execution unit 142 also calculates the pH of the medium at each time during the culture period. The simulation execution unit 142 also calculates the concentration of each protein at each time during the culture period. The simulation execution unit 142 can calculate the amount of each component by a known calculation method. The calculation method is shown in, for example, the literature "Journal of Chemical Technology and Metallurgy, 48, 4, 2013, 351-356 EXPERIMENTAL DETERMINATION OF THE VOLUMETRIC MASS TRANSFER COEFFICIENT". The second storage unit 138 stores the calculation results of the simulation execution unit 142.

[0097] The simulation execution unit 142 determines whether or not the feedback condition is satisfied based on each calculated value at each time. If the feedback condition is satisfied, the simulation execution unit 142 changes some of the culture conditions according to the setting of the feedback condition. For example, the simulation execution unit 142 changes the flow rate data of any of the pumps 98. The simulation execution unit 142 continues the simulation using the changed data. The second storage unit 138 stores the changed condition data.

[0098] When the cell culture simulation is completed, the simulation execution unit 142 calculates the total consumption amount and the total waste amount of the medium in the simulation. The simulation execution unit 142 also calculates the cost using the total consumption amount of the medium and the unit price of the medium. The second storage unit 138 stores the calculation result of the simulation execution unit 142. When step S2 is completed, the process proceeds to step S3.

[0099] In step S3, the user operates input unit 130 to display the results of the simulation. In response to an instruction from input unit 130, display control unit 144 displays the results of the simulation on display unit 134. Display unit 134 displays result screen 152 shown in Fig. 8 and Fig. 9. When step S3 ends, the process proceeds to step S4.

[0100] In step S4, the user determines whether or not a re-simulation is necessary. If the transition of the calculated value in any of the graphs on the result screen 152 is outside the OK range, it is preferable for the user to change the condition data and execute the simulation again. If a re-simulation is necessary (step S4: YES), the process returns to step S1. On the other hand, if a re-simulation is not necessary (step S4: NO), the process proceeds to step S5.

[0101] In step S5, the user cultures cells using the cell culture device 12. The user operates an input device (not shown) of the cell culture device 12 to set the culture conditions specified in step S1 of FIG. 10. For example, when the control unit 20 of the cell culture device 12 and the simulation unit 132 of the simulation device 14 are connected by a signal line, the control unit 20 acquires condition data of the culture conditions from the second storage unit 138 of the simulation unit 132. The cell culture process will be described later. After the cell culture process is completed, the simulation unit 132 acquires condition data of the culture conditions and growth data of new cells from the control unit 20. The second storage unit 138 stores each data acquired from the control unit 20. When step S5 is completed, the process proceeds to step S6.

[0102] In step S6, the user determines whether or not another simulation is required. Cell culture is performed multiple times. The user gradually increases the scale of the cell culture as the number of cell cultures increases. It is preferable for the user to perform a simulation every time the scale of the cell culture is increased. If another simulation is required (step S6: YES), the process returns to step S1. On the other hand, if another simulation is not required (step S6: NO), the cell culture ends.

[0103] 11 is a flow chart showing the flow of cell culture performed using the cell culture device 12. A series of steps shown in FIG. 11 is performed in step S5 shown in FIG.

[0104] In step S11, the control unit 20 performs seeding. The pump control unit 122 controls each pump 98 as shown in FIG. 12. The clamp control unit 124 controls each clamp 100 as shown in FIG. 12. The control unit 20 controls the first supply unit 22a to supply the cell liquid to the first supply flow path 56. Then, the cell liquid is introduced from the first supply unit 22a through the first supply flow path 56 to the first junction 68 of the first circulation flow path 58. The cell liquid introduced to the first junction 68 flows from the first inlet port 48 through the first region 44 and is led to the first outlet port 50. In the first region 44, the cells in the cell liquid adhere to the inner surface of each hollow fiber membrane 40 of the bioreactor 30.

[0105] In step S12, the control unit 20 starts cell culture. Specifically, the control unit 20 controls the first supply unit 22a to supply the culture medium to the first supply flow path 56. Then, the culture medium is introduced from the first supply unit 22a through the first supply flow path 56 to the first junction 68 of the first circulation flow path 58. The culture medium introduced to the first junction 68 circulates through the annular flow path including the first circulation flow path 58, the first inlet port 48, the first region 44, and the first outlet port 50.

[0106] The control unit 20 also controls the second supply unit 22b to supply the basal medium to the second supply flow path 60. The basal medium is then introduced from the second supply unit 22b through the second supply flow path 60 to the second junction 70 of the second circulation flow path 62. The basal medium introduced to the second junction 70 circulates through the annular flow path including the second circulation flow path 62, the second inlet port 52, the second region 46, and the second outlet port 54.

[0107] Furthermore, the gas exchange control unit 126 controls the gas exchange unit 34 to perform gas exchange of the basal medium flowing through the second circulation flow path 62. That is, the gas exchange unit 34 passes a gas of a predetermined component through the basal medium before it flows into the second inlet port 52. This allows the gas concentration (oxygen gas concentration and carbon dioxide gas concentration) and pH of the basal medium introduced into the second inlet port 52 of the bioreactor 30 to be adjusted to values ​​suitable for cell culture. In the bioreactor 30, the culture medium in the first region 44 and the basal medium in the second region 46 are exchanged through the pores of each hollow fiber membrane 40. This allows the gas concentration and pH of the culture medium in the first region 44 to be adjusted.

[0108] Furthermore, the clamp control unit 124 controls the first waste liquid clamp 112 at an appropriate timing to open or close the first waste liquid flow path 76. When the first waste liquid flow path 76 is opened, a part of the culture medium in the first circulation flow path 58 is led to the third waste liquid flow path 80 via the first waste liquid flow path 76. Furthermore, the clamp control unit 124 controls the second waste liquid clamp 114 at an appropriate timing to open or close the second waste liquid flow path 78. When the second waste liquid flow path 78 is opened, a part of the basal medium in the second circulation flow path 62 is led to the third waste liquid flow path 80 via the second waste liquid flow path 78.

[0109] In step S13, the gas sensor 88 measures the oxygen concentration and the carbon dioxide concentration of the medium (culture medium + basal medium). The pH sensor 90 measures the pH of the medium. The gas sensor 88 and the pH sensor 90 transmit the measurement results to the control unit 20. The measurement unit 128 acquires the measurement results from each sensor. The measurement unit 128 stores the acquired measurement results in the first memory unit 120. The gas sensor 88 and the pH sensor 90 perform measurements until the cell culture is completed.

[0110] In step S14, the control unit 20 samples the culture medium. The pump control unit 122 and the gas exchange control unit 126 control the pump (not shown) of the second sampling unit 38 and the clamp (not shown) of the second sampling unit 38 to sample the culture medium in the third waste liquid flow path 80. The sampled culture medium flows into the waste liquid storage unit 26 via the biosensor 92. Here, the first sampling unit 35 may sample the basal culture medium.

[0111] In step S15, the glucose sensor 94 measures the glucose concentration of the medium. The lactate sensor 96 measures the lactate concentration of the medium. The glucose sensor 94 and the lactate sensor 96 transmit the measurement results to the control unit 20. The measurement unit 128 acquires the measurement results from each sensor. The measurement unit 128 stores the acquired measurement results in the first storage unit 120. Here, the concentration of the basal medium sampled by the first sampling unit 35 may be measured.

[0112] In step S16, the control unit 20 cleans the biosensor 92. The second sampling unit 38 is provided with one or more pumps (not shown), one or more clamps (not shown), a cleaning liquid supply unit (not shown), and the like. The pump control unit 122 controls the pump of the second sampling unit 38. Furthermore, the clamp control unit 124 controls the clamp of the second sampling unit 38. Furthermore, the control unit 20 controls the cleaning liquid supply unit. Then, the cleaning liquid flows from the cleaning liquid supply unit to the biosensor 92. As a result, the biosensor 92 is cleaned. The cleaning liquid that has cleaned the biosensor 92 flows into the waste liquid storage unit 26.

[0113] In step S17, the control unit 20 determines whether or not to terminate the cell culture based on the measurement result measured by the biosensor 92. When the control unit 20 determines to terminate the cell culture (step S17: YES), the process proceeds to step S18. On the other hand, when the control unit 20 determines to continue the cell culture (step S17: NO), the process returns to step S14.

[0114] In step S18, the control unit 20 performs cell detachment. The pump control unit 122 turns off the second supply pump 106 and the second circulation pump 108 as shown in FIG. 13. In addition, the clamp control unit 124 controls the first waste liquid clamp 112 and the second waste liquid clamp 114 to close the first waste liquid flow path 76 and the second waste liquid flow path 78 as shown in FIG. 13. In addition, the control unit 20 controls the supply unit 22 to supply the detachment liquid to the first supply flow path 56. Then, the detachment liquid is guided from the supply unit 22 through the first supply flow path 56 and the first circulation flow path 58 to the bioreactor 30. In the bioreactor 30, the detachment liquid detaches the cultured cells from the inner surface of each hollow fiber membrane 40.

[0115] In step S19, the control unit 20 recovers the cells. As shown in Fig. 14, the clamp control unit 124 controls the recovery clamp 110 to open the recovery channel 64. Then, the liquid containing the cells in the first circulation channel 58 is guided to the recovery container 24 via the recovery channel 64. This completes the series of steps of cell culture.

[0116] Incidentally, as in the case of the determination of YES in step S6 in FIG. 10, steps S1 to S6 may be repeated. For example, steps S1 to S6 are performed N times (N≧2). In step S1 for the Nth time, the measurement result in the cell culture process in step S5 for the N-1th time may be used as the proliferation data. In this case, the acquisition unit 140 of the simulation unit 132 acquires data of the measurement result from the first storage unit 120 of the control unit 20. However, each of the glucose measurement result and the lactate measurement result stored in the first storage unit 120 is concentration data. Also, each of the oxygen measurement result and the carbon dioxide measurement result stored in the first storage unit 120 is partial pressure data. When the N-1th measurement result is designated by the input unit 130, the acquisition unit 140 converts the concentration data and the partial pressure data into metabolic rate data. The simulation execution unit 142 uses the converted data to simulate the cell culture.

[0117] [4 Other embodiments] [4-1 Other embodiment 1] Fig. 15 is a diagram showing the configuration of another embodiment of the simulation device 14. As described above, before performing a cell culture simulation, the consumption rate of a protein is calculated. The simulation device 14 shown in Fig. 15 has a function of calculating the consumption rate of a protein.

[0118] The second arithmetic unit 136 also functions as a calculation unit 272. The calculation unit 272 calculates the consumption rate based on the protein concentration input by the input unit 130. The calculation formula is stored in the second storage unit 138 in advance.

[0119] In this case, the user inputs the disappearance rate (deterioration rate, dissolution rate) of each protein and the concentration of each protein in the protein parameter field 182 .

[0120] [4-2 Other embodiment 2] Fig. 16 is a diagram showing the configuration of a simulation system 280. The simulation system 280 shown in Fig. 16 may be used instead of the simulation device 14 shown in Fig. 3. In Fig. 16, the same components as those in Fig. 3 are denoted by the same reference numerals. The simulation system 280 includes at least one first terminal 282, at least one second terminal 284, and a server 286.

[0121] The first terminal 282 may be a personal computer, a smartphone, a tablet, or the like. The first terminal 282 has an input unit 130 and a display unit 134. The first terminal 282 also has a processing circuit and a memory, neither of which are shown. The first terminal 282 is connected to a communication network 288 via a communication device, neither of which are shown.

[0122] A personal computer, a smartphone, a tablet, or the like may be used as the second terminal 284. The second terminal 284 has a control unit 20. The second terminal 284 is connected to a communication network 288 via a communication device (not shown).

[0123] The server 286 includes a simulation unit 132. The server 286 is connected to a communication network 288 via a communication device (not shown). The server 286 may be a cloud server.

[0124] The communication network 288 may be a local area network (LAN) or a wide area network (WAN). The first terminal 282, the second terminal 284, and the server 286 can communicate with each other via the communication network 288.

[0125] When the user operates the input unit 130 to input data, the first terminal 282 transmits each piece of data to the server 286. The server 286 performs a simulation using the data acquired from the first terminal 282. The server 286 transmits the simulation result to the first terminal 282. The first terminal 282 acquires the simulation result from the server 286. The display unit 134 displays the simulation result. The second terminal 284 is capable of acquiring data from the server 286.

[0126] In the simulation system 280, the second calculation unit 136 can also function as a calculation unit 272 shown in FIG.

[0127] Although it depends on the size of the pores of the hollow fiber membrane 40, proteins with large molecular weights (such as fibrinogen, fibronectin, etc.) are less likely to elute from the first region 44 (culture region) to the second region 46 (non-culture region) and are less likely to deteriorate. For such proteins, at least one of the degradation rate and the elution rate does not need to be used when performing a simulation.

[0128] [4-3 Other embodiment 3] In the above-described embodiment, the present invention is used for cell culture using the cell culture device 12 having the hollow fiber membrane 40. However, the present invention can also be used for cell culture that does not use the hollow fiber membrane 40. For example, the present invention can also be used for shaking culture, stirring culture, etc.

Claims

1. A simulation device for simulating cell growth in a cell culture device, comprising: an acquisition unit that acquires a disappearance rate of a predetermined protein in a culture medium independent of cells, a consumption rate of the protein by cells under a first culture condition, and condition data indicating a second culture condition different from the first culture condition; a simulation execution unit that simulates a change in concentration of the protein accompanying cell proliferation under the second culture conditions by using the disappearance rate, the consumption rate, and the condition data acquired by the acquisition unit; a display unit that acquires a concentration of the protein under the second culture conditions as a result of the simulation and displays whether the concentration of the protein falls within a predetermined range; A simulation device comprising:

2. The simulation device according to claim 1 , The cell culture device comprises: A bioreactor; A hollow fiber membrane provided inside the bioreactor; A culture area located in an inner hole of the hollow fiber membrane; A non-culture region located inside the bioreactor and outside the hollow fiber membrane; A first supply unit that supplies each of a culture medium containing the protein and cells to the culture region; a second supply unit that supplies the protein-free basal medium to the non-culture region; A simulation device, wherein the disappearance rate includes a degradation rate of the protein and an elution rate of the protein from the culture area to the non-culture area.

3. The simulation device according to claim 1 or 2, The simulation device further comprises an input unit for inputting the disappearance rate and the consumption rate.

4. The simulation device according to claim 1 or 2, an input unit for inputting the disappearance rate and the change in concentration of the protein measured in cell culture performed under the first culture condition; a calculation unit that calculates the consumption rate based on the concentration of the protein input by the input unit.

5. A simulation system for simulating cell growth in a cell culture device, comprising: an acquisition unit that acquires a disappearance rate of a predetermined protein in a culture medium independent of cells, a consumption rate of the protein by cells under a first culture condition, and condition data indicating a second culture condition different from the first culture condition; a simulation execution unit that simulates a change in concentration of the protein accompanying cell proliferation under the second culture conditions by using the disappearance rate, the consumption rate, and the condition data acquired by the acquisition unit; a display unit that acquires a concentration of the protein under the second culture conditions as a result of the simulation and displays whether the concentration of the protein falls within a predetermined range; A simulation system comprising:

6. The simulation system according to claim 5, A terminal device and a server capable of communicating with each other via a communication network, The terminal device has the display unit, The server includes the acquisition unit and the simulation execution unit.

7. A simulation method for simulating cell growth in a cell culture device, comprising: an acquisition step of acquiring condition data indicating a disappearance rate of a predetermined protein in a medium independent of cells, a consumption rate of the protein by cells under a first culture condition, and a second culture condition different from the first culture condition; a simulation execution step of simulating a change in the concentration of the protein accompanying cell proliferation under the second culture conditions by using the disappearance rate, the consumption rate, and the condition data acquired in the acquisition step; a display step of acquiring a concentration of the protein under the second culture conditions as a result of the simulation, and displaying whether the concentration of the protein falls within a predetermined range; A simulation method comprising:

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