Digital microfluidic cell culture method and digital microfluidic cell culture system
Through digital microfluidic cell culture methods and systems, the problems of static environment and operation complexity in existing DMF cell culture are solved, dynamic cell culture and automated operations are realized, and the growth environment and experimental efficiency of cells are improved.
Patent Information
- Application Number
- CN202411786709.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-06
- Publication Date
- 2025-03-28
AI Technical Summary
The existing DMF cell culture process has limitations in static culture and a lack of dynamic environment, which leads to a lack of fluidity and dynamic exchange of cells under static conditions, affecting cell behavior, metabolic activity and cell-interaction. At the same time, protein adsorption problems and operational complexity also exist.
Through digital microfluidic cell culture methods and systems, the electrode array of the microfluidic chip is used to control the movement of droplets to achieve dynamic culture. The system includes a host computer, a power module, a wireless communication module, a programmable voltage source, a multi-channel switch and a digital microfluidic chip, which can automatically control the introduction and discharge of droplets while meeting the conditions of nutrient supply and metabolic waste discharge.
It realizes continuous nutrient supply and waste liquid discharge to cells, provides a more stable and ideal growth environment, extends the culture cycle, and improves the survival rate and vitality of cells. At the same time, automated operations reduce human errors and improve the efficiency and repeatability of experiments.
Smart Images
Figure CN119709400A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of digital microfluidic technology, and in particular to a digital microfluidic cell culture method and a digital microfluidic cell culture system. Background Art
[0002] Digital microfluidics (DMF) is an advanced technology that precisely manipulates micron-sized droplets through the electrowetting effect, with high flexibility and programmability. In the field of biotechnology, especially in cell culture, DMF has shown great potential. Unlike traditional microfluidics, DMF does not require complex microchannel structures, but controls the movement of droplets on an electrode array through an external electric field to achieve operations such as droplet generation, merging, and splitting. This technology can not only precisely manipulate nanoliter droplets, but also significantly reduce the consumption of samples and reagents, making it very suitable for high-throughput screening and single-cell analysis. Although there are still challenges in biocompatibility, system integration, and automation of complex operations, with the development of materials science and micro-nano manufacturing technology, DMF has broad application prospects in precision medicine, personalized diagnosis, and research on complex biological systems.
[0003] In the existing DMF cell culture process, it is first necessary to design a chip suitable for the experimental needs and perform hydrophobic treatment on its surface to prevent protein adsorption, usually using hydrophobic materials such as Teflon. The electrowetting effect and the applied electric field are used to accurately control the movement of droplets containing the cell suspension and transport them to the predetermined culture position. For suspended cells, the droplets can be suspended in the oil layer on the chip, while adherent cells can achieve attachment and growth through specific surface hydrophilic treatment, such as adhesion patches. When the droplets reach the culture position, the chip is removed and placed in a cell culture incubator for static culture.
[0004] However, there are limitations of static culture in the current DMF cell culture process: lack of a dynamic environment; under static culture conditions, cells in droplets lack fluidity and dynamic exchange, and may not simulate the real cell environment in vivo. This limits the study of cell behavior, metabolic activity, and cell-to-cell interactions. Static culture may lead to poor gas exchange, especially in closed microfluidic chips, which may affect cell growth and metabolism. Protein adsorption problem: Although surface hydrophobic treatment can reduce protein adsorption, hydrophobic materials may still not be able to completely prevent protein adsorption in the presence of long-term use or high concentrations of protein. This may lead to reduced surface activity, thereby affecting the cell culture effect. Operational complexity: The process of removing the chip and placing it in a cell culture incubator may introduce external contamination or interference, affecting the reliability of the culture results. Summary of the invention
[0005] Based on this, it is necessary to address the technical problem of poor DMF cell culture effect in the prior art and propose a digital microfluidic cell culture method and a digital microfluidic cell culture system.
[0006] In a first aspect, a digital microfluidic cell culture method is provided, the method comprising:
[0007] Applied to a digital microfluidic cell culture system, the digital microfluidic cell culture method comprises:
[0008] In response to a first control signal of the microfluidic chip, a growth area is activated, and a pretreated cell suspension is dripped into the growth area, wherein the growth area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located;
[0009] When the nutrient supply conditions are met, the microfluidic chip is controlled to separate fresh culture medium droplets in the preset first electrode array area from the culture medium storage area in the microfluidic chip, and the fresh culture medium droplets are moved to the growth area along a predetermined path;
[0010] When the metabolic waste discharge conditions are met, the microfluidic chip is controlled to separate the droplets in the preset second electrode array area from the growth area and guide them to the waste liquid area in the microfluidic chip.
[0011] In a second aspect, a digital microfluidic cell culture system is provided, comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, the digital microfluidic cell culture system further comprising: a host computer, a power module, a wireless communication module, a programmable voltage source, a multi-way switch, and a digital microfluidic chip, wherein the host computer is used to send a control signal to the processor via the wireless communication module, the processor is used to control the digital microfluidic chip based on the control signal and the multi-way switch, the power module is used to power the wireless communication module, the processor, and the programmable voltage source, the programmable voltage source is used to power the multi-way switch, and the processor implements the steps of the above-mentioned digital microfluidic cell culture method when executing the computer program.
[0012] In a third aspect, a computer-readable storage medium is provided, wherein the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the steps of the above-mentioned digital microfluidic cell culture method are implemented.
[0013] The digital microfluidic cell culture method proposed in the present invention activates the growth area by responding to the first control signal of the microfluidic chip, and drips the pretreated cell suspension into the growth area, wherein the growth area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located, and then when the nutrient supply condition is met, the microfluidic chip is controlled to separate fresh culture medium droplets in the preset first electrode array area from the culture medium storage area in the microfluidic chip, and the fresh culture medium droplets are moved to the growth area along a predetermined path, and finally when the metabolic waste discharge condition is met, the microfluidic chip is controlled to separate droplets in the preset second electrode array area from the growth area, and guide them to the waste liquid area in the microfluidic chip. Fresh culture medium can be introduced from the storage area, and waste liquid can be discharged regularly. This continuous nutrient supply and waste liquid removal mechanism avoids nutrient depletion and waste accumulation, thereby providing a more stable and ideal growth environment for cells, extending the culture cycle, and improving cell survival rate and vitality. Automated nutrient supplementation, waste liquid discharge and remote monitoring greatly reduce the workload of experimenters and improve the efficiency of experimental operation. In particular, dynamic culture can significantly speed up the experimental process when large-scale, high-throughput cell culture is required. In addition, the system is wirelessly controlled through wireless communication and powered by a power module, which completely avoids the complexity and limitations of cable connections in traditional systems. This design makes the system more portable and flexible, suitable for various experimental environments, especially in cell culture boxes or small spaces. Due to the use of wireless communication and battery power, the entire system can operate independently in the incubator for a long time without interference from external equipment, ensuring the stability and continuity of the experiment. In addition, the high efficiency and long battery life of the power module provide reliable energy support for long-term experiments. Through wireless control, users can remotely adjust system parameters, monitor droplet operations and culture processes in real time, thereby realizing a fully automated, unattended cell culture process. This integrated wireless and dynamic control technology greatly improves the system's operational convenience and experimental efficiency, and is widely used in cell research, personalized medicine, drug screening and other fields. BRIEF DESCRIPTION OF THE DRAWINGS
[0014] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0015] in:
[0016] Figure 1 A digital microfluidic cell culture system of a digital microfluidic cell culture method in an embodiment;
[0017] Figure 2 A flowchart of a digital microfluidic cell culture method in one embodiment;
[0018] Figure 3 A schematic diagram of dynamic cell culture in a digital microfluidic cell culture method in one embodiment. DETAILED DESCRIPTION
[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by technicians in the technical field of the present application; the terms used in the specification of the application herein are only for the purpose of describing specific embodiments and are not intended to limit the present application; the terms "including" and "having" and any variations thereof in the specification and claims of the present application and the above-mentioned drawings are intended to cover non-exclusive inclusions. The terms "first", "second", etc. in the specification and claims of the present application or the above-mentioned drawings are used to distinguish different objects, not to describe a specific order.
[0020] Reference to "embodiments" herein means that a particular feature, structure, or characteristic described in conjunction with the embodiments may be included in at least one embodiment of the present application. The appearance of the phrase in various locations in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments. It is explicitly and implicitly understood by those skilled in the art that the embodiments described herein may be combined with other embodiments.
[0021] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0022] The digital microfluidic cell culture method provided by the embodiment of the present invention can be applied in Figure 1In the wireless digital microfluidic cell culture system, the digital microfluidic cell culture system includes a host computer, a power module, a wireless communication module, a programmable voltage source, a multi-way switch and a digital microfluidic chip. The power module uses a lithium battery. The output end of the host computer and the input end of the main control unit are connected through a wireless communication module, the output end of the main control unit is connected to the input end of the multi-way switch, and the output end of the multi-way switch is connected to the input end of the digital microfluidic chip through an FPC connector. The control software running on the host computer is an interface program based on C++ language developed using QT Creator. The main control unit refers to the processor, which can use the STM32H743VI H6 chip to process the control signal, and the wireless communication module uses ESP32 to communicate with the host computer via Bluetooth. The voltage output module uses the MAX1771 chip to achieve voltage boost and uses the HV582 chip to achieve parallel output of multiple signals, and finally controls the electrodes of the digital microfluidic chip through the FPC connector.
[0023] See also Figure 2 As shown, Figure 2 A schematic flow chart of a digital microfluidic cell culture method provided in one embodiment of the present invention includes the following steps:
[0024] Step S101: in response to a first control signal of the microfluidic chip, activating a growth area, and dropping a pretreated cell suspension into the growth area, wherein the growth area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located;
[0025] The digital microfluidic chip consists of a top plate, a bottom plate and a flexible printed circuit (FPC). The top plate is a glass substrate with an indium tin oxide (ITO) coating, and the bottom plate contains a series of driving electrodes, with a 300nm silicon nitride film (SiNx) on the electrode surface as a dielectric layer. The ITO glass and the electrode array substrate are bonded, and the electrode surface is coated with AF Teflon material to meet the hydrophobic requirements, so as to accurately control the movement of droplets and realize efficient digital microfluidic control of complex biological experiments.
[0026] Specifically, refer to Figure 3 , through the control software of the host computer, select the 10×10 electrode array in the digital microfluidic chip, activate the corresponding electrodes, and prepare for the guidance operation of the cell droplets. The pretreated cell suspension is added to the digital microfluidic chip with a 20μL pipette, and the droplet position corresponds to the activated electrode area. Then, by controlling the switch state of the electrode, the droplets are accurately guided to the growth area of the microfluidic chip, ensuring that the CHO-S cells in the droplets are evenly distributed in the growth area, providing a suitable culture environment for cell growth and reproduction.
[0027] In one embodiment, the step before step S101 includes:
[0028] Step A1: Add 5 mL of CHO-S cell suspension with a concentration of 1×10^6 cells / mL into a 15 mL centrifuge tube;
[0029] Step A2: Place the centrifuge tube in a centrifuge, set the centrifugal operation parameters to 1000 rpm, and centrifuge for 5 minutes to precipitate the cells to the bottom of the tube. After the centrifugation is completed, remove the supernatant from the centrifuge tube and retain the cell precipitate;
[0030] Step A3: Use FreeStyle medium as the basal culture medium, supplement with 8 mM glutamine and 30 mM HEPES buffer to prepare a complete culture medium;
[0031] Step A4: Preheat the complete culture medium in a 37°C water bath, and use the preheated complete culture medium to resuspend the centrifuged cells, so that the concentration of the cell suspension droplets is adjusted to 3×10^5 cels / mL to obtain pretreated cell suspension droplets.
[0032] Specifically, in this embodiment, first, 5 mL of a CHO-S cell suspension at a concentration of 1×10^6ce lls / mL was added to a 15 mL centrifuge tube. The centrifuge tube was placed in a centrifuge, the centrifugation parameters were set to 1000 rpm, and centrifuged for 5 minutes to precipitate the cells to the bottom of the tube. After the centrifugation, the supernatant was removed and the cell pellet was retained. FreeStyle medium was used as the basal culture medium, supplemented with 8mM glutamine and 30mM HEPES buffer to prepare a complete culture medium. The complete culture medium was preheated in a 37°C water bath to ensure that the temperature of the cell environment was consistent and to reduce the stress response of the cells to temperature changes. The centrifuged cells were resuspended using the preheated complete culture medium to adjust the concentration of the cell suspension to 3×10^5ce lls / mL.
[0033] In one embodiment, the step before step S101 includes:
[0034] Step B: in response to the second control signal of the microfluidic chip, activating the culture medium storage area, and dripping the prepared complete culture medium into the culture medium storage area, wherein the culture medium storage area refers to the area where the electrode array of the microfluidic chip corresponding to the second control signal is located;
[0035] In this embodiment, the host computer selects the 20×10 electrode array in the microfluidic chip to activate it, and uses a 20μL pipette to drip the prepared complete culture medium into the activated electrode area as the culture medium storage area. On the digital microfluidic chip, the complete culture medium is stored in a specific area as a reserve of nutrient solution required for long-term cell culture. Through the switching of the high-voltage drive circuit and the multi-way switch, the system can dynamically control the release and movement of the culture medium.
[0036] Step S102: when the nutrient supply condition is met, the microfluidic chip is controlled to separate fresh culture medium droplets in the preset first electrode array area from the culture medium storage area in the microfluidic chip, and the fresh culture medium droplets are moved along a predetermined path to the growth area;
[0037] The preset first electrode array area may be an area corresponding to 2×2 electrode arrays.
[0038] As an example, the time information of the growing area is recorded, and whether the nutrient supply condition is satisfied is determined based on the time information. For example, it is determined whether the nutrient supply condition is satisfied every 20 minutes.
[0039] Specifically, to ensure that cells receive a continuous supply of nutrients, and due to the metabolic activity of cells during growth, the nutrients in the culture medium will be consumed, and some metabolic waste will also be produced. If not handled in time, these wastes will have an adverse effect on cell growth. Every 20 minutes, a 2×2 electrode array of fresh culture medium droplets are separated from the storage area. Through microfluidic path planning, the droplets are moved along a predetermined path to the cell growth area and merged with existing droplets in the growth area.
[0040] Step S103: when the metabolic waste discharge conditions are met, the microfluidic chip is controlled to separate the droplets in the preset second electrode array area from the growth area, and guide the droplets to the waste liquid area in the microfluidic chip.
[0041] The preset second electrode array area may be an area corresponding to 2×2 electrode arrays.
[0042] As an example, the time information of the growing area is recorded, and whether the nutrient supply condition is satisfied is determined based on the time information. For example, it is determined whether the nutrient supply condition is satisfied every 20 minutes.
[0043] Specifically, in order to prevent the aging of the culture medium and the accumulation of metabolic waste, a droplet of a 2×2 electrode array is separated from the growth area every 20 minutes, representing the aged culture medium containing metabolic waste. The droplet is guided to the waste liquid area for collection and treatment through the path planning of the microfluidic chip. This method can effectively isolate metabolic waste from the growth environment and keep the cell growth environment clean and healthy. In this way, fresh nutrients can be continuously provided to the cells to maintain the optimal state of the culture environment. The entire droplet manipulation process is automatically controlled by the host computer software, and the user can flexibly adjust the separation frequency, volume and movement path of the droplet through the software interface. The coordinated use of high-voltage drive circuits and multi-way switches realizes the precise manipulation of droplets on the microfluidic chip. Through automated droplet manipulation, regular renewal of the culture medium and automatic discharge of waste liquid are achieved, which significantly improves the automation and repeatability of the experiment and reduces the errors introduced by human operation. Its dynamic adjustment characteristics meet the needs of long-term cell culture and complex biomedical experiments, and provide efficient and reliable solutions for various cell research and biomedical applications.
[0044] It should be noted that the present invention aims to provide a digital microfluidic cell culture method, which is applied to a digital microfluidic cell culture system, and the entire system is placed in a cell culture box. During the culture process, the DMF technology is used to flexibly control the splitting, merging and movement of the droplets, realize the automatic replacement of the culture medium and the automatic addition of reagents, and ensure that the cells continue to grow in a suitable environment. In addition, the chip can be integrated with a microscopic imaging module for real-time monitoring of cell morphological changes, proliferation and metabolic activities. By continuously replenishing and replacing the culture medium, the protein adsorption problem is effectively solved. When the culture is completed, the droplets can be guided to move again by the electric field to achieve cell collection, which is convenient for subsequent analysis, such as gene sequencing, protein expression or functional testing. This process has a high degree of flexibility, automation and accuracy, significantly reduces reagent consumption and operation time, and can also achieve high-throughput cell screening and complex biological experiments, which is particularly suitable for personalized medicine and drug development.
[0045] The present invention has the following advantages: Wireless control: Remote wireless control of the system is achieved through Bluetooth communication, which reduces the dependence on cable connections in traditional systems and improves portability and flexibility. It is particularly suitable for operation in restricted or closed environments, such as cell culture boxes or small laboratory spaces. Lithium battery power supply: The use of lithium batteries to power the system completely avoids the limitation of external power cords, allowing the system to operate independently for a long time, ensuring the continuity and stability of the experiment in a controlled environment, and is particularly suitable for cell experiments that require long-term cultivation. Fully automated, unattended cell culture: Users can adjust parameters and monitor the culture process in real time through Bluetooth to achieve fully automated, unattended operation. This feature greatly improves the convenience and efficiency of the experiment. Dynamic control: The system can dynamically manipulate droplets to achieve precise cell operation and culture process control. This function is the core advantage of digital microfluidics technology and can improve the accuracy and effect of cell culture. Wide applicability: Due to its wireless control and independent power supply capabilities, this invention has broad application potential in fields such as cell research, personalized medicine, and drug screening, and can effectively improve the operating efficiency and research quality of the laboratory. Experimental stability and continuity: Wireless design and lithium battery power supply ensure that the system can run for a long time without external interference, avoiding the problem of experimental interruption or data loss. Droplet manipulation and row and column electrode control: HV582 analog switch is used to control the row and column electrodes, and the droplets are driven to move on the digital microfluidic chip in a time-sharing manner to accurately manipulate biological samples. Continuous nutrient supply and waste liquid discharge: The dynamic culture system can regularly introduce fresh culture medium from the storage area and regularly discharge the waste liquid. This continuous nutrient supply and waste liquid removal mechanism avoids nutrient depletion and waste accumulation, thereby providing cells with a more stable and ideal growth environment, extending the culture cycle, and improving cell survival rate and vitality. High degree of automation: Through programming control and wireless communication modules, dynamic culture realizes fully automated cell culture operations. The system can automatically adjust the frequency and amount of culture medium replenishment and waste liquid removal according to preset parameters, reducing manual intervention, reducing human operation errors, and improving the repeatability and reliability of the experiment. Real-time remote monitoring and control: Thanks to the wireless communication module (ESP32) and Bluetooth connection, the dynamic culture system can achieve remote real-time monitoring and control. Researchers can remotely adjust the culture conditions through the host computer or mobile device, which is especially useful in long-term culture or restricted environments (such as biosafety cabinets). Precise droplet manipulation: The dynamic culture system achieves precise control of droplets by time-sharing control of row and column electrodes. During the culture process, the movement of culture medium and waste liquid is program-controlled, which ensures high precision of droplet manipulation, avoids mixing or cross-contamination during droplet manipulation, and ensures the accuracy of experimental results.Dynamic regulation of cell growth environment: Dynamic culture allows the voltage and time of droplet movement to be adjusted through voltage source and potentiometer programming control, which can dynamically adjust the culture conditions and provide the specific environment required by cells at different growth stages. This flexible environmental regulation helps to optimize cell growth rate and state. Improve experimental efficiency: Automated nutrient supplementation, waste liquid discharge and remote monitoring greatly reduce the workload of experimental personnel and improve experimental operation efficiency. In particular, dynamic culture can significantly speed up the experimental process when large-scale, high-throughput cell culture is required.
[0046] In one embodiment, a digital microfluidic cell culture system is provided. The digital microfluidic cell culture system includes a processor, a memory, a network interface, a database, a host computer, a power module, a wireless communication module, a programmable voltage source, a multi-way switch and a digital microfluidic chip connected through a system bus, wherein the host computer is used to send a control signal to the processor through the wireless communication module, the processor is used to control the digital microfluidic chip based on the control signal and the multi-way switch, the power module is used to power the wireless communication module, the processor and the programmable voltage source, and the programmable voltage source is used to power the multi-way switch. Wherein, the processor of the digital microfluidic cell culture system is used to provide computing and control capabilities. The memory of the digital microfluidic cell culture system includes a non-volatile and / or volatile storage medium and an internal memory. The non-volatile storage medium stores an operating system, a computer program and a database. The internal memory provides an environment for the operation of the operating system and the computer program in the non-volatile storage medium. The network interface of the digital microfluidic cell culture system is used to communicate with an external client through a network connection. When the computer program is executed by the processor, a function or step of a digital microfluidic cell culture method is realized. The system can be wirelessly controlled via Bluetooth communication and powered by lithium batteries, completely avoiding the complexity and limitations of cable connections in traditional systems. This design makes the system more portable and flexible, suitable for various experimental environments, especially in cell culture boxes or small spaces. Due to the use of wireless communication and battery power, the entire system can operate independently in the incubator for a long time without interference from external equipment, ensuring the stability and continuity of the experiment. In addition, the high efficiency and long battery life of lithium batteries provide reliable energy support for long-term experiments. Through Bluetooth control, users can remotely adjust system parameters, monitor droplet operations and culture progress in real time, thereby realizing a fully automated, unattended cell culture process. This integrated wireless, dynamic control technology greatly improves the system's operational convenience and experimental efficiency, and is widely used in cell research, personalized medicine, drug screening and other fields.
[0047] In one embodiment, a digital microfluidic cell culture system is provided, comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor implements the following steps when executing the computer program:
[0048] In response to a first control signal of the microfluidic chip, a growth area is activated, and a pretreated cell suspension is dripped into the growth area, wherein the growth area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located;
[0049] When the nutrient supply conditions are met, the microfluidic chip is controlled to separate fresh culture medium droplets in the preset first electrode array area from the culture medium storage area in the microfluidic chip, and the fresh culture medium droplets are moved to the growth area along a predetermined path;
[0050] When the metabolic waste discharge conditions are met, the microfluidic chip is controlled to separate the droplets in the preset second electrode array area from the growth area and guide them to the waste liquid area in the microfluidic chip.
[0051] In one embodiment, a computer-readable storage medium is provided, wherein the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the following steps are implemented:
[0052] In response to a first control signal of the microfluidic chip, a growth area is activated, and a pretreated cell suspension is dripped into the growth area, wherein the growth area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located;
[0053] When the nutrient supply conditions are met, the microfluidic chip is controlled to separate fresh culture medium droplets in the preset first electrode array area from the culture medium storage area in the microfluidic chip, and the fresh culture medium droplets are moved to the growth area along a predetermined path;
[0054] When the metabolic waste discharge conditions are met, the microfluidic chip is controlled to separate the droplets in the preset second electrode array area from the growth area and guide them to the waste liquid area in the microfluidic chip.
[0055] It should be noted that the functions or steps that can be implemented by the computer-readable storage medium or the digital microfluidic cell culture system can be correspondingly referred to the relevant descriptions on the server side and the client side in the aforementioned method embodiment. To avoid repetition, they will not be described one by one here.
[0056] Those of ordinary skill in the art can understand that all or part of the processes in the above-mentioned embodiment methods can be completed by instructing related hardware through a computer program, and the computer program can be stored in a non-volatile computer-readable storage medium. When the computer program is executed, it can include the processes of the embodiments of the above-mentioned methods. Among them, any reference to memory, storage, database or other media used in the embodiments provided in this application may include non-volatile and / or volatile memory. Non-volatile memory may include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM) or flash memory. Volatile memory may include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM is available in many forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDRSDRAM), enhanced SDRAM (ESDRAM), synchronous link DRAM (SLDRAM), RAMbus direct RAM (RDRAM), direct RAMbus dynamic RAM (DRDRAM), and RAMbus dynamic RAM (RDRAM), etc.
[0057] Those skilled in the art can clearly understand that for the convenience and simplicity of description, only the division of the above-mentioned functional units and modules is used as an example. In actual applications, the above-mentioned functions can be distributed and completed by different functional units and modules as needed, that is, the internal structure of the device can be divided into different functional units or modules to complete all or part of the functions described above.
[0058] The embodiments described above are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. Such modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention, and should all be included in the protection scope of the present invention.
Claims
1. A digital microfluidic cell culture method, characterized in that: Applied to a digital microfluidic cell culture system, the digital microfluidic cell culture system further comprises: a host computer, a power module, a wireless communication module, a processor, a programmable voltage source, a multi-way switch and a digital microfluidic chip, wherein the host computer is used to send a control signal to the processor through the wireless communication module, the processor is used to control the digital microfluidic chip based on the control signal and the multi-way switch, the power module is used to supply power to the wireless communication module, the processor and the programmable voltage source, and the programmable voltage source is used to supply power to the multi-way switch; The digital microfluidic cell culture method comprises: In response to a first control signal of the microfluidic chip, a growth area is activated, and a pretreated cell suspension is dripped into the growth area, wherein the growth area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located; When the nutrient supply conditions are met, the microfluidic chip is controlled to separate fresh culture medium droplets in the preset first electrode array area from the culture medium storage area in the microfluidic chip, and the fresh culture medium droplets are moved to the growth area along a predetermined path; When the metabolic waste discharge conditions are met, the microfluidic chip is controlled to separate the droplets in the preset second electrode array area from the growth area and guide them to the waste liquid area in the microfluidic chip.
2. The digital microfluidic cell culture method according to claim 1, characterized in that: The step of activating the growth area in response to the first control signal of the microfluidic chip and dropping the pretreated cell suspension into the growth area, wherein the first electrode area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located, comprises: In response to the second control signal of the microfluidic chip, the culture medium storage area is activated, and the prepared complete culture medium is dripped into the culture medium storage area, wherein the culture medium storage area refers to the area where the electrode array of the microfluidic chip corresponding to the second control signal is located.
3. The digital microfluidic cell culture method according to claim 1, characterized in that: The step of activating the growth area in response to the first control signal of the microfluidic chip and dropping the pretreated cell suspension into the growth area, wherein the first electrode area refers to the area where the electrode array of the microfluidic chip corresponding to the first control signal is located, comprises: Add 5 mL of CHO-S cell suspension with a concentration of 1×10^6 cel ls / mL into a 15 mL centrifuge tube; Place the centrifuge tube in a centrifuge, set the centrifugal operating parameters to 1000 rpm, and centrifuge for 5 minutes to precipitate the cells to the bottom of the tube. After the centrifugation is completed, remove the supernatant from the centrifuge tube and retain the cell precipitate; FreeStyle medium was used as the basal culture medium, supplemented with 8 mM glutamine and 30 mM HEPES buffer to prepare a complete culture medium; The complete culture medium was preheated in a 37°C water bath, and the centrifuged cells were resuspended in the preheated complete culture medium to adjust the concentration of the cell suspension droplets to 3×10^5 cells / mL to obtain pretreated cell suspension droplets.
4. A digital microfluidic cell culture system, comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, characterized in that: The digital microfluidic cell culture system further includes: a host computer, a power module, a wireless communication module, a programmable voltage source, a multi-way switch and a digital microfluidic chip, wherein the host computer is used to send a control signal to the processor through the wireless communication module, the processor is used to control the digital microfluidic chip based on the control signal and the multi-way switch, the power module is used to supply power to the wireless communication module, the processor and the programmable voltage source, and the programmable voltage source is used to supply power to the multi-way switch; When the processor executes the computer program, the steps of the digital microfluidic cell culture method according to any one of claims 1 to 3 are implemented.
5. A computer-readable storage medium storing a computer program, characterized in that: When the computer program is executed by a processor, the steps of the digital microfluidic cell culture method according to any one of claims 1 to 3 are implemented.