Cell microsphere culture device based on droplet microfluidic technology
By using a pulse centrifugal pump and a circulating turbulence device in the cell microsphere culture device, the problem of removing oil and surfactants in droplet microfluidics was solved, achieving efficient nutrient exchange and a biomechanical biomimetic microenvironment, providing highly active and functional culture conditions for cells.
Patent Information
- Application Number
- CN202422606794.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-28
- Publication Date
- 2025-12-12
- Estimated Expiration
- 2034-10-28
AI Technical Summary
In the preparation of cell gel microspheres using existing droplet microfluidic technology, the use of oil and surfactants makes subsequent removal difficult, affecting cell viability and making it hard to control nutrient exchange.
A cell microsphere culture device based on droplet microfluidics technology is used. A periodic turbulence is formed in the culture flask by a pulse centrifugal pump and a circulating turbulence device to promote the exchange of nutrients inside and outside the cell microspheres. The flow rate and speed are controlled by adjusting the power of the pulse centrifugal pump.
It improves the efficiency of nutrient diffusion, provides a biomechanical biomimetic microenvironment, ensures the high activity and functionality of organoids, and avoids the adverse effects of oil and surfactants on cells.
Smart Images
Figure CN223660096U_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The utility model relates to cell culture technical field, specifically is a cell microsphere culture device based on droplet microfluidic technology. BACKGROUND
[0002] 2D cultured cells are different from in vivo cells in physiology and cell response. These differences promote the rapid popularization of 3D cell culture technology. More and more evidence shows that 3D cell culture can better represent the in vivo environment and produce more physiological cell models, and even the clinical expression information reflected by the gene expression profile of 3D cultured cells is more accurate than that of 2D cultured cells. The retention rate and survival rate of 3D cells are key problems, and microspheres can provide protection for cells and significantly improve cell transplantation rate as an excellent carrier for cell delivery.
[0003] Droplet microfluidic technology can precisely control the flow of various fluids at the micro-nano scale, so as to generate micro-nano droplets with adjustable structure and controllable composition in a high-throughput manner. By combining suitable hydrogel materials and manufacturing methods, single or multiple cells can be efficiently encapsulated into hydrogels to prepare cell gel microspheres. Cell gel microspheres can provide a three-dimensional, relatively independent and controllable microenvironment for cell proliferation and differentiation.
[0004] Gel microspheres, as an advanced functional material, have important application value in many fields, especially as a cell carrier, which can well simulate the survival environment of cells in vivo and provide a novel three-dimensional culture method for cells. The microspheres generated based on droplet microfluidic technology have become one of the most effective technologies for preparing cell gel microspheres due to their size, structure and composition.
[0005] Since the most common way to prepare gel microspheres is emulsification, the hydrogel solution is mixed with immiscible oil to form droplets, and a surfactant is added to the emulsion system to stabilize the emulsion. The size of the microspheres usually depends on the concentration of the polymer, the emulsification rate and the water / oil volume ratio, therefore, the use of oil and surfactant cannot be avoided. The use of oil makes its subsequent removal troublesome, not to mention that the washing step can cause cell damage and may have adverse effects on cell viability.
[0006] In addition, gel microspheres encapsulate cells in microgels, and the size of the microspheres is fixed by photopolymerization, so the exchange between substances is inevitably more difficult than in the single cell period. UTILITY MODEL CONTENT
[0007] The utility model discloses a cell microsphere culture device based on liquid drop microfluidic technique, to realize through the efficient culture process of optimization organoid, improve the diffusion efficiency of nutrient substance, and provide biomechanics bionic microenvironment for organoid, guarantee the high activity and functional purpose of organoid.
[0008] In order to realize the above-mentioned purpose, the utility model adopts the following technical means:
[0009] A cell microsphere culture device based on liquid drop microfluidic technique, including the culture bottle for holding culture solution, the culture bottle top is placed with circulating torrent device, the circulating torrent device includes the installation box of the bottle mouth of the culture bottle, the bottom surface of the installation box is installed with the operation pipe, the operation pipe is set apart with the interior of the installation box, the operation pipe is installed with the partition plate, the upper portion of the partition plate is as the liquid outlet chamber, the liquid outlet chamber is connected with the culture bottle, the lower portion of the partition plate is as the liquid return chamber, the liquid return chamber is connected with the culture bottle, the installation box is provided with pulse centrifugal pump, the liquid inlet end of the pulse centrifugal pump is communicated circulation pipe, the liquid inlet end of the circulation pipe is inserted into the liquid return chamber, the liquid outlet end of the pulse centrifugal pump is communicated with the liquid outlet pipe, and the liquid outlet pipe is inserted into the liquid outlet chamber.
[0010] As preferred, the installation box includes the control box that abuts the bottle mouth top end of the culture bottle, and the bottom surface of the control box is configured with a spherical block that extends into the culture bottle, wherein the diameter of the spherical block is the same as the diameter of the bottle mouth of the culture bottle.
[0011] Further, the control box is provided with a control unit for driving the pulse centrifugal pump, and the pulse centrifugal pump is arranged in the spherical block.
[0012] Further, the liquid outlet pipe is arranged in the installation box and extends into the liquid outlet chamber through the bottom surface of the installation box, and the circulation pipe is arranged in the installation box and extends into the liquid return chamber by sequentially penetrating through the installation box, the liquid outlet chamber and the partition plate downward.
[0013] Further, the sidewall of the operation pipe is configured with a plurality of liquid outlet through holes that are communicated with the liquid outlet chamber.
[0014] Further, the operation pipe is vertically arranged, and the bottom end of the operation pipe is configured with a liquid return port that is communicated with the liquid return chamber.
[0015] In the process of using the utility model, the following beneficial effects are achieved:
[0016] The cell microsphere culture is carried out in the culture bottle, the working pipe is extended into the culture solution during the culture process, and the outlet cavity and the return cavity are both extended below the liquid level of the culture solution. Then the pulse centrifugal pump is started, under the action of the pulse centrifugal pump, the culture solution in the culture bottle is continuously pumped out through the circulating pipe, then the culture solution is periodically pumped into the outlet cavity, and finally the culture solution is periodically introduced into the culture bottle through the outlet cavity, a periodic turbulent flow is formed, the cell microspheres in the culture bottle are stirred, and the exchange of nutrients inside and outside the cells is promoted. And the flow and speed can be adjusted by adjusting the power of the pulse centrifugal pump. BRIEF DESCRIPTION OF DRAWINGS
[0017] Figure 1 It is a structural schematic diagram of the utility model.
[0018] Figure 2 It is a sectional structure schematic diagram. Figure 1
[0019] It is a front view structure schematic diagram. Figure 3 Figure 2
[0020] Among them, 1-culture bottle, 2-installation box, 3-working pipe, 4-separation plate, 5-outlet cavity, 6-return cavity, 7-pulse centrifugal pump, 8-circulating pipe, 9-outlet pipe, 10-control box, 11-spherical block, 12-outlet through hole, 13-return port. DETAILED DESCRIPTION
[0021] In order to make the purpose, technical scheme and advantages of the embodiments of the utility model clearer, the technical scheme in the embodiments of the utility model will be described clearly and completely in combination with the drawings in the embodiments of the utility model. Obviously, the described embodiments are part of the embodiments of the utility model, rather than all the embodiments. The components of the embodiments of the utility model described and shown in the drawings can be arranged and designed in various different configurations.
[0022] Therefore, the following detailed description of the embodiments of the utility model provided in the drawings is not intended to limit the scope of the claimed utility model, but only represents selected embodiments of the utility model. Based on the embodiments in the utility model, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the utility model.
[0023] It should be noted that the embodiments in the utility model and the features in the embodiments can be combined with each other without conflict.
[0024] It should be noted that like numerals and letters refer to like items throughout the several views, and once an item is defined in one view, it should not require further defining and explaining in subsequent views.
[0025] In the description of the utility model, it needs to explain, the term "center", "upper", "lower", "left", "right", "vertical", "horizontal", "internal", "external" and so on indicate the orientation or position relation is based on the orientation or position relation shown in the drawing, or it is the orientation or position relation that the utility model product uses usually, or it is the orientation or position relation that the person skilled in the art usually understands, just is for the convenience of describing the utility model and simplifying the description, and therefore can not be understood as indicating or implying that the indicated device or element must have a particular orientation, a particular orientation and operation, therefore can not be understood as the limitation of the utility model.In addition, the terms "first", "second" and so on are only used for distinguishing description, and can not be understood as indicating or implying relative importance.
[0026] In the description of the utility model, it also needs to explain that, unless otherwise explicitly specified and limited, the terms "set", "mount", "connect", "connect" should be understood broadly, for example, it can be fixedly connected, can be detachably connected, or integrally connected, can be mechanically connected, or electrically connected, can be directly connected, or indirectly connected through intermediate medium, can be the communication inside two elements.The above-mentioned terms in the utility model can be understood according to the specific meaning of the person skilled in the art.
[0027] Please refer to Figures 1 to 3 As shown in the figure, a cell microsphere culture device based on droplet microfluidic technology, including culture bottle 1 for containing culture solution, the top of the culture bottle 1 is placed with circulating flow device, the circulating flow device includes installation box 2 placed in the bottle mouth of the culture bottle 1, the bottom surface of the installation box 2 is installed with operation pipe 3, the operation pipe 3 is inwardly partitioned with the installation box 2, the operation pipe 3 is installed with partition plate 4, the upper portion of the partition plate 4 is used as liquid outlet cavity 5, the liquid outlet cavity 5 is communicated with the culture bottle 1, the lower portion of the partition plate 4 is used as liquid return cavity 6, the liquid return cavity 6 is communicated with the culture bottle 1, the installation box 2 is provided with pulse centrifugal pump 7, the liquid inlet end of the pulse centrifugal pump 7 is communicated with circulating pipe 8, the liquid inlet end of the circulating pipe 8 extends into the liquid return cavity 6, the liquid outlet end of the pulse centrifugal pump 7 is communicated with liquid outlet pipe 9, and the liquid outlet end of the liquid outlet pipe 9 extends into the liquid outlet cavity 5.
[0028] In this way, the cell microsphere culture is carried out in the culture bottle 1, the working pipe 3 is extended into the culture solution during the culture process, and the outlet cavity 5 and the return cavity 6 are both extended below the liquid level of the culture solution. Then, the pulse centrifugal pump 7 is started, and under the action of the pulse centrifugal pump 7, the culture solution in the culture bottle 1 is continuously pumped out through the circulation pipe 8, then the culture solution is periodically pumped into the outlet cavity 5, and finally the culture solution is periodically introduced into the culture bottle 1 through the outlet cavity 5, so as to form a periodic turbulent flow, make the cell microspheres in the culture bottle 1 surge, and promote the exchange of nutrients inside and outside the cells. And the flow and speed can be adjusted by adjusting the power of the pulse centrifugal pump 7.
[0029] At the same time, the outlet cavity 5 is located above the return cavity 6, so as to avoid the influence of the turbulent flow generated during the culture process on the state of the culture solution entering the return cavity 6.
[0030] Further, the mounting box 2 comprises a control box 10 abutting against the top end of the bottle mouth of the culture bottle 1, and the bottom surface of the control box 10 is provided with a spherical block 11 extending into the culture bottle 1, and the diameter of the spherical block 11 is the same as the diameter of the bottle mouth of the culture bottle 1.
[0031] In this way, by arranging the spherical block 11, the spherical block 11 can abut against the inner wall of the bottle mouth of the culture bottle 1, so as to avoid the mounting box 2 from shaking back and forth when abutting against the top end of the bottle mouth of the culture bottle 1.
[0032] Further, the control box 10 is provided with a control unit for driving the pulse centrifugal pump 7, and the pulse centrifugal pump 7 is arranged in the spherical block 11.
[0033] Further, the outlet pipe 9 is arranged in the mounting box 2 and extends into the outlet cavity 5 through the bottom surface of the mounting box 2, and the circulation pipe 8 is arranged in the mounting box 2 and extends into the return cavity 6 by sequentially penetrating the mounting box 2, the outlet cavity 5 and the partition plate 4 downwards.
[0034] In this way, the integration of the pulse centrifugal pump 7, the outlet pipe 9 and the circulation pipe 8 is higher, so as to avoid that the space in the culture bottle 1 is occupied too much and affects the volume of the culture solution that can be loaded.
[0035] At the same time, the side wall of the working pipe 3 is provided with a plurality of outlet through holes 12 communicating with the outlet cavity 5.
[0036] In this way, by arranging the outlet through holes 12 on the side wall of the working pipe 3, the horizontal turbulent flow can be generated when the culture solution passes through, and then the generated turbulent flow moves towards the inner wall of the culture bottle 1, so as to make the turbulent flow interact with the inner wall of the culture bottle 1, and further improve the disturbance to the culture solution.
[0037] Meanwhile, the operation pipe 3 is vertically arranged, and a bottom end of the operation pipe 3 is configured with a liquid return port 13 in communication with the liquid return cavity 6.
[0038] Although the utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement to part of the technical features, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the utility model shall be included in the protection scope of the utility model.
Claims
1. A cell microsphere culture device based on droplet microfluidic technology, characterized in that, The application relates to a culture bottle (1) for containing culture solution, wherein a circulating jet device is arranged on the top of the culture bottle (1), the circulating jet device comprises a mounting box (2) arranged on the bottle mouth of the culture bottle (1), a working pipe (3) is arranged on the bottom surface of the mounting box (2) and is inwardly partitioned from the mounting box (2), a partition plate (4) is arranged in the working pipe (3), an outflow cavity (5) is arranged above the partition plate (4) and is communicated with the culture bottle (1), a return flow cavity (6) is arranged below the partition plate (4) and is communicated with the culture bottle (1), a pulse centrifugal pump (7) is arranged in the mounting box (2), the inlet of the pulse centrifugal pump (7) is communicated with a circulating pipe (8), the inlet of the circulating pipe (8) extends into the return flow cavity (6), and the outlet of the pulse centrifugal pump (7) is communicated with an outflow pipe (9) whose outlet extends into the outflow cavity (5).
2. The cell microsphere culture device based on droplet microfluidic technology according to claim 1, characterized in that, The mounting box (2) comprises a control box (10) which is abutted against the top end of the bottle mouth of the culture bottle (1), the bottom surface of the control box (10) is provided with a spherical block (11) which extends into the culture bottle (1) and has the same diameter as the diameter of the bottle mouth of the culture bottle (1). 3.The cell microsphere culture device based on the droplet microfluidic technology according to claim 2, characterized in that, The control box (10) is provided with a control unit for driving the pulse centrifugal pump (7), and the pulse centrifugal pump (7) is arranged in the spherical block (11).
4. The cell microsphere culture device based on droplet microfluidic technology according to claim 1, characterized in that, The outflow pipe (9) is arranged in the mounting box (2) and extends into the outflow cavity (5) through the bottom surface of the mounting box (2), and the circulating pipe (8) extends into the return flow cavity (6) by sequentially penetrating the mounting box (2), the outflow cavity (5) and the partition plate (4) from top to bottom.
5. The cell microsphere culture device based on droplet microfluidic technology according to claim 1, characterized in that, The side wall of the working pipe (3) is provided with a plurality of outflow through holes (12) which are communicated with the outflow cavity (5).
6. The cell microsphere culture device based on droplet microfluidic technology according to claim 1, characterized in that, The working pipe (3) is vertically arranged, and the bottom end of the working pipe (3) is provided with a return flow port (13) which is communicated with the return flow cavity (6).