Cell culture auxiliary device for cell culture
By using a modularly designed cell culture auxiliary device, the traditionally dispersed cell processing steps are integrated into a continuous operation system, realizing the automation and high efficiency of the cell culture process. This solves the problem of low efficiency in traditional methods, improves cell separation purity and survival rate, and reduces energy consumption and pollution risks.
Patent Information
- Application Number
- CN202511714606.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-21
- Publication Date
- 2026-03-20
AI Technical Summary
In existing technologies, cell culture processes are fragmented, resulting in low efficiency and slow workflow.
Design a cell culture auxiliary device that integrates traditionally dispersed cell processing steps into a continuous operation system through modular design. Utilize components such as electric clamp valves, 22-channel diverting valves, flow meter sensors, magnetic bead sorting devices, and peristaltic pumps to achieve automated control. Combine magnetic bead sorting and centrifugation technologies to achieve efficient cell separation and culture.
It significantly improves the efficiency of cell culture, shortens the cell passage cycle, reduces the risk of contamination, increases cell survival rate and separation purity, and reduces energy consumption and magnetic bead recovery costs.
Smart Images
Figure CN121699741A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of cell culture, in particular to a cell culture auxiliary device for cell culture. BACKGROUND
[0002] Cell culture refers to a method of simulating the in-vivo environment (sterile, suitable temperature, pH and certain nutritional conditions, etc.) to make it survive, grow, reproduce and maintain the main structure and function. Cell culture is also called cell cloning technology, and its formal name in biology is cell culture technology. Cell culture is an essential process for both the entire biological engineering technology and the biological cloning technology, and cell culture itself is a large-scale cloning of cells. Cell culture technology can make a cell become a simple single cell or a few differentiated multicellular through mass culture, which is an essential part of cloning technology, and cell culture itself is cell cloning. Cell culture device is a core equipment in cell biology, biological engineering and medical research, which can be divided into laboratory and industrial levels according to application scenarios and scales, and its functions include environment simulation, sterile operation and cell growth monitoring. The present application provides a cell culture auxiliary device for cell culture, which integrates traditional dispersed cell processing links into a continuous operation system. SUMMARY
[0003] (I) Technical problems to be solved In view of the defects of the prior art, the present application provides a cell culture auxiliary device for cell culture, which has the advantages of integrating traditional dispersed cell processing links into a continuous operation system, and solves the problems of dispersed cell culture process, slow cell culture workflow and low work efficiency.
[0004] (II) Technical solutions To achieve the above-mentioned purpose, the present application provides the following technical solutions: a cell culture auxiliary device for cell culture, comprising a material bag, a motor-operated pinch valve connected below the material bag, a 22-channel diverter valve arranged below the motor-operated pinch valve, a flowmeter sensor arranged below the left side of the 22-channel diverter valve, a magnetic bead sorting device arranged below the flowmeter sensor, a peristaltic pump arranged on the right side of the flowmeter sensor, an 8-channel diverter valve arranged below the magnetic bead sorting device, three 8-channel diverter valves arranged below the magnetic bead sorting device, a waste liquid collection module arranged below the magnetic bead sorting device, a magnetic bead cleaning module arranged on the right side of the waste liquid collection module, a centrifugal structure arranged on the right side of the magnetic bead cleaning module, and a culture unit connected on the right side of the peristaltic pump.
[0005] Preferably, the magnetic bead sorting device is close to the magnetic column by permanent magnet, the magnetic beads to be passed are adsorbed on the magnetic column, the specific antibody (such as anti-biotin microbead) coupled on the surface of the magnetic beads is combined with the target cell surface antigen to form a complex, the superparamagnetic microbead (about 50 nm in diameter) has no damage to the cell and is biodegradable, the magnetic field generated by the permanent magnet magnetizes the magnetic beads, a high gradient magnetic field is formed on the surface of the magnetic column to adsorb the magnetic bead-cell complex, and the unlabeled cells flow out of the magnetic column.
[0006] Preferably, the centrifugal structure comprises a centrifugal device, which separates the target substance from a complex mixture by centrifugal differential sedimentation or buoyancy balance, and realizes separation by using the difference in sedimentation speed of different density components in the mixture in the centrifugal field. The formula for calculating the centrifugal force (RCF) is: RCF = 1.118 x 10-5 x r x (rpm)2; Wherein r is the radius of rotation (cm), and rpm is the rotation speed.
[0007] Preferably, the flow meter sensor collects the instantaneous flow of the fluid at a certain moment and the total flow of the cumulative flow in a period of time, ensures the accuracy of the data, and the flow meter sensor 4 collects the instantaneous flow of the fluid at a certain moment and the total flow of the cumulative flow in a period of time, ensures the accuracy of the data, and the calibration and verification process of the flow meter is as follows: Pre-calibration: use a standard device with a precision of ≤1.5% to adjust the flow meter parameters to ensure that the instantaneous flow error is ≤2%; Post-sampling verification: verify the deviation of the actual flow from the set value, and the allowable error is ≤5% (such as fluoride sampling); Environmental compensation technology: integrate temperature sensor (such as ±0.1℃ precision) and pressure sensor to dynamically compensate parameters such as sound speed and fluid density.
[0008] Preferably, the peristaltic pump pumps fluid by alternately squeezing and releasing the elastic hose of the pump, the motor drives the rollers (usually 3-6) in the pump head to rotate, the rollers successively squeeze the hose to form a "pillow" shaped fluid section, and the single squeezing volume is determined by the inner diameter of the hose and the geometric parameters of the rollers. The self-rotation design of the rollers can reduce the friction loss with the hose.
[0009] Preferably, the 8-channel steering valve controls left and right rotation by a stepper motor to realize liquid flow, the valve body structure adopts an 8-channel valve body, the internal flow channel is optimized by CFD, and the pressure drop is <0.1 MPa @ 10 L / min. The sealing technology adopts PTFE composite material sealing between the valve core and the valve sleeve, the pressure rating reaches 35 MPa, the leakage is <0.1 mL / min, the controller sends a pulse sequence (such as 200 Hz) to the stepper motor driver, the subdivision driving mode (such as 16 subdivisions) improves the positioning accuracy, the feedback signal (such as a Hall sensor) realizes closed-loop control, and the position error is <0.1°. When a fault protection occurs, the overcurrent protection circuit cuts off the power supply when locked, preventing the motor from burning out.
[0010] Preferably, the culture unit comprises upper dynamic culture and lower static culture, simulates and maintains a specific controllable environment, provides stable "artificial living conditions" for the growth, reproduction, metabolism or experiment of biological samples such as cells, microorganisms, tissues and embryos, and ensures the accuracy, repeatability and activity of experimental results; the PID algorithm is used to adjust pH (7.2-7.4), dissolved oxygen content (>95% saturation) and temperature (±0.1℃ fluctuation).
[0011] Compared with the prior art, the application provides a cell culture auxiliary device for cell culture, which has the following beneficial effects: 1. The cell culture auxiliary device realizes full-process automatic control through modular design, and the core innovation point is to integrate traditional dispersed cell processing links into a continuous operation system. The device takes a material bag as a starting end, accurately controls the opening and closing of fluid by an electric pinch valve, realizes intelligent distribution of multiple culture media / reagents by a 22-channel steering valve. The flow sensor sensor monitors the flow rate in real time and forms a closed-loop control with the peristaltic pump, and the magnetic bead separation device adopts electromagnetic field and fluid mechanics cooperative technology, which can complete high-purity separation of target cells within 3 minutes. The specially designed 8-channel steering valve array cooperates with three independent waste liquid collection modules to improve the waste liquid treatment efficiency by 60%, and the magnetic bead cleaning module realizes magnetic bead recovery rate ≥95% through gradient centrifugation technology. The parallel layout of the centrifugal structure and the culture unit shortens the cell subculture period to 1 / 3 of the traditional method.
[0012] 2. The whole system realizes seamless connection from cell sorting to expansion culture through PLC programming, significantly reduces the pollution risk of manual operation. The device has the following advantages in practical application: the linkage system of the 22-channel steering valve and the flow sensor makes the culture medium distribution accuracy reach ±0.5%, which is 90% lower than the error of traditional manual operation; the magnetic bead separation device optimizes the electromagnetic field parameters to realize target cell separation purity ≥98%, and the magnetic bead residual amount is controlled at 0.1 μg / 10 6The cell is below. Its modular design compresses the cell passage time from the conventional 4 hours to 1.5 hours, and the waste liquid treatment module can synchronously treat 8 channels of waste liquid, so that the treatment efficiency is improved, in the stem cell culture experiment, the survival rate of the cells maintained by the device is improved, and the pollution rate caused by the frequent opening and closing of the traditional culture system is reduced from 15% to below 3%, in addition, the gradient centrifugation technology of the magnetic bead cleaning module reduces the cost of magnetic bead recovery, and the annual operation energy consumption of the whole system is lower than that of similar equipment. BRIEF DESCRIPTION OF DRAWINGS
[0013] Figure 1 It is a general schematic diagram of the cell culture device of the application. Figure 2 It is a structure schematic diagram of the magnetic bead sorting device of the application. Figure 3 It is a centrifugal structure schematic diagram of the application. Figure 4 It is a flow meter sensor structure schematic diagram of the application. Figure 5 It is a peristaltic pump structure schematic diagram of the application. Figure 6 It is an 8-channel directional valve structure schematic diagram of the application. Figure 7 It is a culture unit schematic diagram of the application.
[0014] 1, material bag; 2, electric pinch valve; 3, 22-channel directional valve; 4, flow meter sensor; 5, magnetic bead sorting device; 6, peristaltic pump; 7, waste liquid collection module; 8, magnetic bead cleaning module; 9, centrifugal structure; 10, culture unit; 11, 8-channel directional valve. DETAILED DESCRIPTION
[0015] The technical solutions in the embodiments of the application will be described clearly and completely below with reference to the drawings in the embodiments of the application. Obviously, the described embodiments are only part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the application.
[0016] Reference Figures 1-7The utility model relates to a kind of cell culture auxiliary device for cell culture, including material bag 1, the lower portion of the material bag 1 is connected with electric pinch valve 2, the lower portion of electric pinch valve 2 is provided with 22 channel diverter valve 3, the left lower portion of 22 channel diverter valve 3 is provided with flowmeter sensor 4, the lower portion of flowmeter sensor 4 is provided with magnetic bead sorting device 5, the right side of flowmeter sensor 4 is provided with peristaltic pump 6, the lower portion of magnetic bead sorting device 5 is provided with 8 channel diverter valve 11, 8 channel diverter valve 11 is provided with three, the lower portion of magnetic bead sorting device 5 is provided with waste liquid collection module 7, the right side of waste liquid collection module 7 is provided with magnetic bead cleaning module 8.The right side of magnetic bead cleaning module 8 is provided with centrifugal structure 9, and the right side of peristaltic pump 6 is connected with culture unit 10.
[0017] Further, the magnetic bead sorting device 5 is close to the magnetic column by permanent magnet, the magnetic beads to be passed are adsorbed on the magnetic column, the specific antibody (such as anti-biotin microbeads) on the surface of the magnetic beads is coupled, and the complex is formed by combining with the target cell surface antigen. The superparamagnetic microbeads (about 50 nm in diameter) are non-injurious to cells and biodegradable. The magnetic field generated by the permanent magnet magnetizes the magnetic beads, a high-gradient magnetic field is formed on the surface of the magnetic column, the magnetic bead-cell complex is adsorbed, and the unlabeled cells flow out of the magnetic column. The magnetic field generated by the permanent magnet magnetizes the magnetic beads, a high-gradient magnetic field is formed on the surface of the magnetic column, the magnetic bead-cell complex is adsorbed, and the unlabeled cells flow out of the magnetic column.
[0018] Further, the centrifugal structure 9 comprises a centrifugal device, which separates the target substance from a complex mixture by centrifugal differential sedimentation or buoyancy balance, and separates the target substance from the complex mixture by using the difference in sedimentation speed of different density components in the centrifugal force field. The formula for calculating the centrifugal force (RCF) is: RCF=1.118×10-5×r×(rpm)2; Wherein r is the radius of rotation (cm), and rpm is the rotation speed.
[0019] Further, the flowmeter sensor 4 collects the instantaneous flow rate of the fluid at a certain moment and the total flow rate in a certain period of time, ensures the accuracy of the data, and the flowmeter sensor 4 collects the instantaneous flow rate of the fluid at a certain moment and the total flow rate in a certain period of time, ensures the accuracy of the data, and the calibration and verification process of the flowmeter is as follows: Pre-calibration: use a standard device with a precision of ≤1.5% to adjust the flowmeter parameters to ensure that the instantaneous flow error is ≤2%; Post-sampling verification: verify the deviation of the actual flow rate from the set value, and the allowable error is ≤5% (such as fluoride sampling); Environmental compensation technology: integrate temperature sensor (such as ±0.1℃ precision) and pressure sensor to dynamically compensate parameters such as sound speed and fluid density.
[0020] Further, the peristaltic pump 6 pumps fluid by alternately squeezing and releasing the elastic hose of the pump. The motor drives the rollers (usually 3-6) in the pump head to rotate, and the rollers successively squeeze the hose to form a "pillow" shaped fluid section. The volume of a single squeeze is determined by the inner diameter of the hose and the geometric parameters of the rollers. The self-rotating design of the rollers can reduce frictional wear with the hose.
[0021] Further, the 8-channel directional valve 11 controls left and right rotation by a stepper motor to realize liquid flow direction. The valve body structure adopts an 8-channel valve body, and the internal flow channel is optimized by CFD, with a pressure drop of <0.1 MPa at 10 L / min. The sealing technology uses PTFE composite material to seal between the valve core and the valve sleeve, with a pressure rating of up to 35 MPa and a leakage of <0.1 mL / min. By sending a pulse sequence (such as 200 Hz) to the stepper motor driver through the controller, the positioning accuracy is improved by subdividing the driving mode (such as 16 subdivisions). Feedback signals (such as Hall sensors) realize closed-loop control, with a position error of <0.1°. When a fault occurs, the overcurrent protection circuit cuts off the power supply when locked, preventing the motor from burning.
[0022] Further, the culture unit 10 includes upper dynamic culture and lower static culture, simulates and maintains specific controllable environment, provides stable "artificial survival conditions" for the growth, reproduction, metabolism or experiment of biological samples such as cells, microorganisms, tissues and embryos, and ensures the accuracy, repeatability and activity of experimental results. PID algorithm is used to adjust pH (7.2-7.4), dissolved oxygen content (>95% saturation) and temperature (±0.1℃ fluctuation).
[0023] Example one: A cell culture auxiliary device for cell culture, comprising a material bag 1, the lower part of the material bag 1 is connected with an electric pinch valve 2, the lower part of the electric pinch valve 2 is provided with a 22-channel diversion valve 3, the left lower part of the 22-channel diversion valve 3 is provided with a flow meter sensor 4, the lower part of the flow meter sensor 4 is provided with a magnetic bead sorting device 5, the right side of the flow meter sensor 4 is provided with a peristaltic pump 6, the lower part of the magnetic bead sorting device 5 is provided with an 8-channel diversion valve 11, the 8-channel diversion valve 11 is provided with three, the lower part of the magnetic bead sorting device 5 is provided with a waste liquid collection module 7, the right side of the waste liquid collection module 7 is provided with a magnetic bead cleaning module 8. The right side of the magnetic bead cleaning module 8 is provided with a centrifugal structure 9, and the right side of the peristaltic pump 6 is connected with a culture unit 10; the cell culture auxiliary device realizes full-process automatic control through modular design, and the core innovation point is that the traditional dispersed cell processing links are integrated into a continuous operation system. The device takes the material bag as the starting end, accurately controls the opening and closing of the fluid through the electric pinch valve, and realizes the intelligent distribution of multiple culture media / reagents through the 22-channel diversion valve. The flow meter sensor realizes real-time monitoring of the flow rate and linkage with the peristaltic pump to form a closed-loop control, and the magnetic bead sorting device adopts electromagnetic field and fluid mechanics cooperative technology, which can complete the high-purity separation of target cells within 3 minutes. The specially designed 8-channel diversion valve array cooperates with the three independent waste liquid collection modules, so that the waste liquid treatment efficiency is improved by 60%, and the magnetic bead cleaning module realizes the magnetic bead recovery rate of ≥95% through gradient centrifugation technology. The parallel layout of the centrifugal structure and the culture unit shortens the cell subculture cycle to 1 / 3 of the traditional method.
[0024] Example two: The magnetic bead sorting device 5 is close to the magnetic column through a permanent magnet, and the magnetic beads to be passed are adsorbed on the magnetic column. The surface of the magnetic beads is coupled with specific antibodies (such as anti-biotin microbeads), and the specific antibodies are combined with the target cell surface antigen to form a complex. The superparamagnetic microbeads (about 50nm in diameter) are non-injurious to cells and biodegradable. The magnetic field generated by the permanent magnet magnetizes the magnetic beads, and a high-gradient magnetic field is formed on the surface of the magnetic column to adsorb the magnetic bead-cell complex. The unlabeled cells flow out of the magnetic column. The magnetic field generated by the permanent magnet magnetizes the magnetic beads, and a high-gradient magnetic field is formed on the surface of the magnetic column to adsorb the magnetic bead-cell complex. The unlabeled cells flow out of the magnetic column. The centrifugal structure 9 comprises a centrifugal device, which separates target substances from complex mixtures through centrifugal differential sedimentation or buoyancy balance. The centrifugal force (RCF) is calculated by the formula: RCF=1.118×10-5×r×(rpm)2.
[0025] Example three: The flow meter sensor 4 collects the instantaneous flow of the fluid at a certain moment and the total flow in a certain period of time, ensuring the accuracy of the data. The calibration and verification process of the flow meter is as follows: Pre-sampling calibration: use standard device with precision ≤1.5% to adjust flow meter parameters, ensure instantaneous flow error ≤2%; Post-sampling verification: verify the deviation of actual flow from set value, allow error ≤5% (such as fluoride sampling); Environmental compensation technology: integrate temperature sensor (such as ±0.1℃ precision) and pressure sensor to dynamically compensate parameters such as sound speed and fluid density; The peristaltic pump 6 pumps fluid by alternately squeezing and releasing the elastic hose of the pump. The motor drives the rollers (usually 3-6) in the pump head to rotate. The rollers successively squeeze the hose to form a "pillow" shaped fluid section. The single squeezing volume is determined by the inner diameter of the hose and the geometric parameters of the roller. The self-rotation design can reduce the friction loss of the hose.
[0026] Example Four: 8-way directional valve 11 rotates left and right by step motor control, realizing liquid flow direction. The valve body structure adopts 8-way valve body, and the internal flow channel is optimized by CFD, with pressure drop <0.1MPa@10L / min. Sealing technology: PTFE composite material is used for sealing between valve core and valve sleeve, with pressure rating up to 35MPa, leakage <0.1mL / min. By sending pulse sequence (such as 200Hz) to step motor driver through controller, subdivision driving mode (such as 16 subdivision) is used to improve positioning accuracy. Feedback signal (such as Hall sensor) realizes closed-loop control, with position error <0.1°. When fault protection occurs, the overcurrent protection circuit cuts off the power supply when locked rotor, preventing motor burning; The culture unit 10 includes upper dynamic culture and lower static culture, simulates and maintains specific controllable environment, provides stable "artificial living conditions" for the growth, reproduction, metabolism or experiment of biological samples such as cells, microorganisms, tissues and embryos, and ensures the accuracy, repeatability and activity of experimental results; PID algorithm is used to adjust pH (7.2-7.4), dissolved oxygen content (>95% saturation) and temperature (±0.1℃ fluctuation).
[0027] Although embodiments of the present application have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and alterations can be made thereto without departing from the principles and spirit of the present application, the scope of which is defined by the appended claims and their equivalents.
Claims
1. A cell culture auxiliary device for cell culture, comprising a material bag (1), characterized in that: An electric clamp valve (2) is connected to the bottom of the material bag (1). A 22-channel diverting valve (3) is provided below the electric clamp valve (2). A flow meter sensor (4) is provided to the lower left of the 22-channel diverting valve (3). A magnetic bead sorting device (5) is provided below the flow meter sensor (4). A peristaltic pump (6) is provided to the right of the flow meter sensor (4). An 8-channel diverting valve (11) is provided below the magnetic bead sorting device (5). There are three 8-channel diverting valves (11). A waste liquid collection module (7) is provided below the magnetic bead sorting device (5). A magnetic bead cleaning module (8) is provided to the right of the waste liquid collection module (7). A centrifugal structure (9) is provided to the right of the magnetic bead cleaning module (8). A culture unit (10) is connected to the right of the peristaltic pump (6).
2. The cell culture auxiliary device for cell culture according to claim 1, characterized in that: The magnetic bead sorting device (5) uses a permanent magnet to approach the magnetic column and attract the magnetic beads to be passed onto the magnetic column.
3. The cell culture auxiliary device for cell culture according to claim 1, characterized in that: The centrifugal structure (9) includes a centrifugal device that separates the target substance from a complex mixture by centrifugal differential sedimentation or buoyancy balance.
4. A cell culture auxiliary device for cell culture according to claim 1, characterized in that: The flow meter sensor (4) collects the instantaneous flow rate and cumulative flow rate of the fluid in real time to ensure data accuracy.
5. A cell culture auxiliary device for cell culture according to claim 1, characterized in that: The peristaltic pump (6) pumps fluid by alternately squeezing and releasing the pump's elastic hose.
6. A cell culture auxiliary device for cell culture according to claim 1, characterized in that: The 8-channel directional valve (11) is controlled to rotate left and right by a stepper motor to achieve the direction of liquid flow.
7. A cell culture auxiliary device for cell culture according to claim 1, characterized in that: The culture unit (10) includes an upper dynamic culture layer and a lower static culture layer, which simulates and maintains a specific controllable environment to provide stable artificial living conditions for the growth, reproduction, metabolism or experiments of biological samples such as cells, microorganisms, tissues, and embryos, and to ensure the accuracy, repeatability and activity of experimental results.