Fully closed loop smart cell culture system

Abstract

To provide an automated, fully closed-loop smart cell culture system.SOLUTION: A fully closed loop smart cell culture system 1 includes a cell culture device 2 and integrated control means 4. The cell culture device 2 includes a culture means 6 and a detection means 7. The culture means 6 includes a culture medium storage section 61 for storing a culture medium, a cell culture section 64 for accommodating a culture solution mixed with cells to be cultured, and a first pump 62 configured to provide the culture medium from the culture medium storage section 61 to the cell culture section 64. The detection means 7 includes a measurement module 72 and a second pump 71 for supplying the culture solution from the cell culture section 64 to the measurement module 72, and outputs the measurement result of the measurement module 72 for the culture solution. The integrated control means 4 is electrically connected to the cell culture device 2, receives a detection signal from the detection means 7, generates control parameters based on the detection signal, and transmits the control parameters to the cell culture device 2.SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present invention relates to a microbiology device, and in particular to a fully closed loop smart cell culture system. [Background technology]

[0002] In conventional cell culture technology, cells are manually extracted from the cell culture device, their status and quantity are checked, and the culture medium is added or updated based on the confirmed information. However, this manual work is time-consuming as it needs to be repeated many times, and there is also a risk of contamination of the cell culture environment.

[0003] To solve this problem, for example, Patent Document 1 discloses a cell culture device that is configured to have a culture means that contains liquid culture medium from a culture medium supply means, an air supply means that provides the air necessary for cell growth, a pressure control means that provides the necessary pressure environment for the culture means, and a cell number detection means that detects the number of healthy living cells in the culture means.

[0004] However, this conventional technique lacks convenience because it does not have a mechanism for automatically replenishing liquid culture medium based on the detection results after the number of live cells is detected by the cell number detection means. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Taiwan Patent No. I732357B Specification Summary of the Invention [Problem to be solved by the invention]

[0006] Therefore, an object of the present invention is to provide an automated, fully closed-loop, smart cell culture system. [Means for solving the problem]

[0007] In view of the above problems, the present invention provides a full-closed loop smart cell culture system including a cell culture device and an integrated control means, At least one of the cell culture devices has a culture means and a detection means, The culture means includes a culture medium storage unit that stores a liquid culture medium not mixed with the cells to be cultured, a heating unit, a cell culture unit that contains a culture solution in which the cells to be cultured are mixed into the liquid culture medium, and a first pump that is configured to provide the culture medium from the culture medium storage unit to the heating unit to heat it, and then provide it to the cell culture unit; the detection means includes a measurement module and a second pump that supplies the culture solution from the cell culture unit to the measurement module, and is configured to output a measurement result of the culture solution by the measurement module; the integrated control means is electrically connected to the cell culture device, receives the detection signal from the detection means, generates control parameters based on the detection signal, and transmits the control parameters to the cell culture device; The cell culture device provides a full closed loop smart cell culture system, characterized in that the amount of culture medium provided to the cell culture section by the first pump via the heating section is controlled based on the control parameters received from the integrated control means. Effect of the Invention

[0008] The present invention drives a first pump to supply culture medium from a culture medium storage section to a cell culture section based on the state of cells, such as the quantity of cells, detected by the detection means, thereby realizing the provision of an automated, fully closed-loop, smart cell culture system with reduced external intervention in the cell culture device, and suppressing increased costs and the risk of contamination due to manual labor. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 is a perspective view showing an embodiment of a fully closed-loop smart cell culture system of the present invention. [Diagram 2]FIG. [Diagram 3] FIG. 2 is a block diagram showing the configuration of the embodiment. [Figure 4] FIG. 2 is an explanatory diagram showing the connection relationship of the microfluidic chip in the cell culture device of the same embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] 1 to 3 show an embodiment of the present invention, and as shown in the figures, a fully closed-loop smart cell culture system 1 of the present invention includes three cell culture devices 2, a microinjection means 3, an integrated control means 4, and a touch panel 5. Each cell culture device 2 is configured to be capable of culturing one type of cell, and examples of cells that can be cultured by the fully closed-loop smart cell culture system 1 of the present invention include, but are not limited to, NK (natural killer) cells and T cells.

[0011] Each cell culture device 2 includes a culture means 6 and a detection means 7. The culture means 6 is electrically connected to the integrated control means 4, and includes a culture medium storage unit 61, a first pump 62, a heating unit 63, a cell culture unit 64, and an environment management module 65. The culture medium storage unit 61 includes a first container 611 that contains a liquid culture medium (not shown) that is not mixed with cells to be cultured. The heating unit 63 includes a heating container 631 that communicates with the first container 611 and temporarily contains unheated culture medium supplied from the first container 611, and two heating modules (not shown) that are arranged, for example, on both sides of the heating container 631. The cell culture unit 64 includes a cell culture container 641 that communicates with the heating container 631 and contains heated culture medium and cells to be cultured supplied from the heating container 631, and a shaking module 642 that holds the cell culture container 641 and shakes it to uniformly distribute the cells to be cultured contained in the cell culture container 641. In the following description, the liquid in which the cells to be cultured are mixed in the liquid medium in the cell culture vessel 641 is referred to as the culture medium.

[0012] The environment management module 65 has a temperature sensor (not shown) disposed in the medium storage section 61 to detect the temperature around the medium storage section 61, a temperature control section (not shown) to adjust the temperature around the medium storage section 61, another temperature sensor (not shown) disposed in the cell culture section 64 to detect the temperature around the cell culture section 64, another temperature control section (not shown) to adjust the temperature around the cell culture section 64, a gas sensor (not shown) disposed in the cell culture section 64 to detect the concentration of carbon dioxide and other gases around the cell culture section 64, and a pH meter (not shown) disposed in the cell culture vessel 641 to detect the pH value of the culture solution. In this way, the environment management module 65 monitors and adjusts the temperature around the medium storage section 61, the temperature and gas components around the cell culture section 64, and the pH value of the culture solution so that the culture environment is suitable for the cells to be cultured.

[0013] Specifically, for example, the environmental temperature of the medium storage unit 61 is preferably 4°C±2°C, the environmental temperature of the cell culture unit 64 is preferably 37°C±1°C, the CO2 concentration around the cell culture unit 64 is preferably 5%±1%, and the pH value of the culture solution is preferably set to pH7.0 to pH7.6. The environmental management module 65 transmits data on the detected temperatures, gas components, pH value, etc. of the medium storage unit 61 and the cell culture unit 64 to the integrated control means 4, and the integrated control means 4 controls the operation of the environmental management module 65 and other necessary devices based on the detection results.

[0014] In this embodiment, a tube pump is used as the first pump 62, the culture medium is supplied from the first container 611 to the heating container 631, the culture medium contained in the heating container 631 is heated by the heating module, and the heated culture medium is then supplied by the first pump 62 from the heating container 631 to the cell culture container 641 of the cell culture section 64. The shaking module 642 shakes the cell culture container 641 to uniformly mix the newly supplied culture medium with the specimen cells that were in the cell culture container 641, and to uniformly distribute the cells to be cultured contained in the cell culture container 641.

[0015] As shown in Figures 2 to 4, the detection means 7 is electrically connected to the integrated control means 4, and has a second pump 71, a detection module 72, a buffer solution storage section 73, a detection valve module 74, and a waste liquid storage tank 75.

[0016] In this embodiment, a tube pump is adopted as the second pump 71, which supplies the culture fluid from the cell culture container 641 to the detection module 72. The detection module 72 has a microfluidic chip 76 and an optical member 77. The microfluidic chip 76 surrounds a micropassage 764 through which the culture fluid can pass, and the micropassage 764 communicates with the second pump 71 via a first inlet 761, communicates with the micro injection means 3 via a second inlet 762 and a third inlet 763, and communicates with the waste liquid storage tank 75 via an outlet 765.

[0017] The optical member 77 includes a microscope 771 , a camera 772 , and an image processor 773 electrically connected to the camera 772 .

[0018] The buffer solution storage section 73 communicates with the second pump 71 and has a buffer solution container 731 that contains a buffer solution (not shown).

[0019] The detection valve module 74 is configured to control the communication state between the buffer solution container 731 and the second pump 71 , and the communication state between the second pump 71 and the microfluidic chip 76 .

[0020] Incidentally, in this embodiment, the first container 611, the heating container 631, the cell culture container 641, and the buffer container 731 possessed by each cell culture device 2 can be configured in a bag shape, a case shape, or a dish shape; in other words, as long as they can appropriately contain the liquid to be contained, their shapes are not limited.

[0021] The microinjection means 3 is electrically connected to the integrated control means 4 and includes a first microinjector 31 communicating with the second inlet 762 of the microfluidic chip 76 of each cell culture device 2, a buffer tank 32 communicating with the first microinjector 31 and containing a buffer solution (not shown), a second microinjector 33 communicating with the third inlet 763 of the microfluidic chip 76 of each cell culture device 2, a staining agent storage tank 34 communicating with the second microinjector 33 and containing a staining agent (not shown), and a drive motor 35 electrically connected to the integrated control means 4. The drive motor 35 simultaneously drives the first microinjector 31 and the second microinjector 33 using a screw mechanism, so that the first microinjector 31 and the second microinjector 33 simultaneously supply the buffer solution from the buffer tank 32 and the staining agent from the staining agent storage tank 34 into the micropassage 764, respectively.

[0022] For each cell culture device 2, the integrated control means 4 starts the second pump 71 to supply a part of the culture solution from the cell culture container 641 into the micropassage 764 via the first inlet 761 of the microfluidic chip 76, and then drives the first microsyringe 31 using the drive motor 35 to supply the buffer solution from the buffer tank 32 into the micropassage 764 via the second inlet 762, and sends the culture solution into the micropassage 764. In addition, the second microsyringe 33, which is simultaneously driven by the drive motor 35, supplies the dye from the dye storage tank 34 into the micropassage 764 via the third inlet 763, thereby dyeing the cells to be cultured contained in the culture solution and making them easier to observe.

[0023] The camera 772 photographs the culture solution in the micropassage 764 through the microscope 771, generates several pieces of cell image data, and outputs them to the image processor 773. The image processor 773 generates detection results by performing image recognition on these cell image data, and outputs detection signals reflecting these detection results to the integrated control means 4. Here, the image recognition performed by the image processor 773 on these cell image data means observing the state and number of the cells to be cultured contained in the culture solution in the micropassage 764, thereby generating detection results including the survival rate, density, and number of live cells of the cells to be cultured.

[0024] Then, the integrated control means 4 continues to control the drive motor 35 to drive the first micro-syringe 31 and the second micro-syringe 33 to simultaneously supply the buffer solution and the staining agent into the micro-passage 764, respectively, and further pump the culture solution into the micro-passage 764 until the culture solution enters the waste liquid storage tank 75 from the outlet 765.

[0025] Here, by adjusting the driving force from drive motor 35 to adjust the speed at which first microsyringe 31 and second microsyringe 33 provide the buffer solution and dye agent into micropassage 764, the proceeding speed of the culture medium through micropassage 764 can be controlled, making it easier for camera 772 to capture images of micropassage 764.

[0026] The integrated control means 4 receives a detection signal from the cell culture device 2, generates control parameters based on the detection result contained in the detection signal and a processing rule set in advance, and transmits a control signal containing this control parameter to the corresponding cell culture device 2. The cell culture device 2 drives the first pump 62 based on this control parameter, and adjusts the additional amount of culture medium supplied from the first container 611 to the cell culture container 641 via the heating container 631.

[0027] Regarding the processing rule here, several control modes are selectable, and for example, the additional amount of culture medium to be added to the cell culture vessel 641 can be calculated based on a countermeasure table set in advance, or can be set according to the user's judgment. As an example of the countermeasure table, for example, the following Table 1 can be referred to. That is, when the volume of the culture medium in the cell culture vessel 641 is 10 ml and the number of cells to be cultured is unknown, a part of the culture medium is supplied from the cell culture vessel 641 to the detection module 72 by the second pump 71 and the detection result is obtained that the culture medium in the cell culture vessel 641 contains 2,000,000 cells to be cultured. Based on this detection result, the integrated control means 4 calculates 10 ml as the additional amount of culture medium to be added from the culture medium storage unit 61 to the cell culture vessel 641 in order to return the amount of the culture medium in the cell culture vessel 641 to 20 ml, and accordingly controls the first pump 62 to supply 10 ml of culture medium from the first vessel 611 to the cell culture vessel 641 and mix it.

[0028] [Table 1]

[0029] When the integrated control means 4 receives a detection signal from the cell culture device 2, it drives the second pump 71 of the corresponding cell culture device 2 to inject the buffer solution from the buffer solution container 731 into the first inlet 761 of the microfluidic chip 76 via the detection valve module 74, and causes the buffer solution to flow into the micro passage 764 until it is discharged from the discharge port 765 into the waste liquid storage tank 75. Therefore, each cell culture device 2 can clean the detection valve module 74 and the micro passage 764 by sending the buffer solution from the buffer solution container 731 into the micro passage 764 and discharging the culture solution remaining in both the detection valve module 74 and the micro passage 764, thereby preventing them from being affected in the next detection.

[0030] After receiving the detection signal from the cell culture device 2, the integrated control means 4 can automatically add culture medium to the cell culture device 2 and simultaneously perform the cleaning operation of the detection valve module 74 and the micro passage 764.

[0031] In this embodiment, the three cell culture devices 2 of the fully closed-loop smart cell culture system 1 are integrated into the same housing, and the medium storage section 61 and the cell culture section 64 of each of these cell culture devices 2 are grouped together in the same location.

[0032] Furthermore, the types of cells cultured by each of the three cell culture devices 2 in the full-closed-loop smart cell culture system 1 may be the same or different. The same type of cells can also be cultured under different culture conditions. The control status for each cell culture device 2 does not need to be the same. For example, when detecting the number of cells for one cell culture device 2, it is also possible to add culture medium and wash the detection valve module 74 and the micropassage 764 for the other cell culture devices 2.

[0033] The touch panel 5 is electrically connected to the integrated control means 4, and displays information such as the environmental temperature of the medium storage section 61 in each cell culture device 2, the environmental temperature of the cell culture section 64, the concentration of CO2 and other gases, and the pH value of the culture solution contained in the cell culture vessel 641, or image data of the culture solution in the micro passage 764 captured by the camera 772. This allows the user to monitor the cell culture status. In addition, the user can set processing rules corresponding to each cell culture device 2 and parameters of the culture environment to the integrated control means 4 via the touch panel 5. In addition, the integrated control means 4 can also support remote control through communication means such as the Internet. This allows the user to take appropriate action through a mobile device such as a smartphone, even if he or she cannot access the touch panel 5.

[0034] In summary, the full-closed-loop smart cell culture system 1 of the present invention uses each cell culture device 2 to match the culture means 6 and the detection means 7, thereby enabling cells to be cultured in the cell culture vessel 641 of the culture means 6, and the second pump 71 of the detection means 7 supplies a part of the culture fluid in the cell culture vessel 641 to the microfluidic chip 76, and the first micro-injector 31 and the second micro-injector 33 control the movement of the culture fluid in the micro-passage 764. In addition, the camera 772 takes an image of the culture fluid, and the image processor 773 generates a detection result corresponding to the data of the cells contained in the culture fluid. Furthermore, the integrated control means 4 is used to judge the detection result and generate a control parameter based on the processing rule, so that the first pump 62 can be controlled based on this control parameter to adjust the additional amount added from the first vessel 611 to the cell culture vessel 641. Therefore, the full-closed-loop smart cell culture system 1 of the present invention realizes the provision of an automated full-closed-loop smart cell culture system by reducing external intervention in the cell culture device, and can suppress the increase in cost and the risk of contamination due to human labor.

[0035] Although the embodiment of the present invention has been described above, the present invention is not limited to this, and various modifications are possible without departing from the gist of the present invention. [Explanation of symbols]

[0036] 1. Fully closed-loop smart cell culture system 2 Cell culture device 3. Microinjection methods 31 The first micro syringe 32 Buffer tank 33 Second Micro Syringe 34 Dyeing agent tank 35 Drive motor 4. Integrated Control Means 5. Touch Panel 6 Culture means 61 Culture medium storage section 611 First Container 62 First Pump 63 Heating section 631 Heating container 64 Cell Culture Department 641 Cell culture vessels 642 Shaking Module 65 Environmental Management Module 7. Detection Methods 71 Second Pump 72 Inspection module 73 Buffer Reservoir 731 Buffer container 74 Inspection valve module 75 Waste liquid storage tank 76 Microfluidic Chips 761 First Entrance 762 Second Entrance 763 Third Entrance 764 Micro Passage 765 Outlet 77 Optical Components 771 Microscope 772 Camera 773 Image Processor

Claims

1. A fully closed loop smart cell culture system comprising a cell culture device and an integrated control means, At least one of the cell culture devices has a culture means and a detection means, The culture means includes a culture medium storage unit that stores a liquid culture medium not mixed with the cells to be cultured, a heating unit, a cell culture unit that contains a culture solution in which the cells to be cultured are mixed into the liquid culture medium, and a first pump that is configured to supply the culture medium from the culture medium storage unit to the heating unit, heat the culture medium, and then supply the culture medium to the cell culture unit; the detection means includes a measurement module and a second pump that supplies the culture solution from the cell culture unit to the measurement module, and is configured to output a measurement result of the culture solution by the measurement module; the integrated control means is electrically connected to the cell culture device, receives a detection signal from the detection means, generates control parameters based on the detection signal, and transmits the control parameters to the cell culture device; the cell culture device controls the amount of the culture medium to be provided to the cell culture unit via the heating unit by the first pump based on the control parameters received from the integrated control means; a micro-injection means electrically connected to said integrated control means; The detection module of the cell culture device includes a microfluidic chip having a microchannel through which the culture solution can pass, and an optical member that calculates the number of the cells in the culture solution by observing the culture solution passing through the microchannel; The second pump supplies the culture medium from the cell culture unit to the microfluidic chip; A fully closed loop smart cell culture system, characterized in that the microinjection means has a first microinjector that communicates with the micropassage and delivers the culture medium into the micropassage.

2. 2. The full-closed-loop smart cell culture system according to claim 1, wherein the microinjection means comprises: a staining agent storage tank for storing a staining agent; and a second microinjector communicating with the staining agent storage tank and the micropassage and supplying the staining agent from the staining agent storage tank to the micropassage, thereby staining the culture solution in the micropassage.

3. The detection means of the cell culture device a buffer storage unit for storing a buffer and communicating with the second pump; and a detection valve module for controlling a communication state between the buffer storage unit and the second pump and a communication state between the second pump and the microfluidic chip; the integrated control means controls operation of the second pump to provide the buffer solution from the buffer solution reservoir to the microfluidic chip via the detection valve module; The full-closed-loop smart cell culture system according to claim 1, characterized in that the buffer solution is pumped into the micro-passage to wash the detection valve module and the micro-passage of the microfluidic chip.

4. The culture means of the cell culture device is electrically connected to the integrated control means, and controls the temperature around the medium storage unit, the temperature around the cell culture unit, and CO ２ 2. The full-closed-loop smart cell culture system according to claim 1, further comprising an environment control module for detecting and controlling the concentration and pH value of the culture solution.

5. 2. The full-closed-loop smart cell culture system according to claim 1, wherein the cell culture unit of the cell culture device comprises: a cell culture vessel that contains the culture medium; and a shaking module that shakes the cell culture vessel to uniformly distribute the cells in the culture medium contained in the cell culture vessel.

6. The full-closed loop smart cell culture system according to claim 1, characterized in that the culture medium storage unit of the cell culture device has a first container for accommodating the culture medium, and the heating unit has a heating container for accommodating and heating the culture medium provided from the first container.

7. The full-closed loop smart cell culture system according to claim 1 , wherein the first pump and the second pump are both tube pumps.

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