A double tangential flow microfluidic exosome separation device

Abstract

The utility model relates to biomedical engineering technology related fields especially a kind of double tangential flow microfluidic exosome separation device, including centrifugal component, ultrafiltration component, liquid inlet pipeline and collection component, the centrifugal component is connected with ultrafiltration component by liquid inlet pipeline, the collection component is set in the liquid outlet of ultrafiltration component, the centrifugal component includes centrifugal barrel, centrifugal motor and lower base, the centrifugal barrel is installed in the output end of centrifugal motor, the centrifugal motor is fixed on lower base, the collection component includes exosome collection bottle, supernatant collection bottle and shunt valve, the exosome collection bottle and supernatant collection bottle are connected with ultrafiltration component by shunt valve, can realize the efficient accurate separation to exosome, it is convenient to reduce the loss of active ingredient in separation process, improve the recovery rate and purity of exosome, it is convenient to simplify the operation process of exosome separation, reduce the error caused by artificial intervention, improve the repeatability and reliability of experiment.

Description

Technical Field

[0001] This utility model relates to the field of biomedical engineering technology, and in particular to a dual-tangential flow microfluidic exosome separation device. Background Technology

[0002] In the field of lung disease treatment, traditional treatment methods have significant limitations. Oral or injected drugs not only have low utilization rates and are prone to causing systemic side effects, but also have difficulty in accurately targeting damaged lung sites, thus limiting the therapeutic effect. In recent years, the technology of nebulized inhalation of mesenchymal stem cell exosome culture supernatant has become a research hotspot, providing a new direction for lung health management. Exosomes secreted by mesenchymal stem cells during the culture process serve as the core medium for intercellular communication, carrying bioactive molecules such as proteins, mRNA, and miRNA. The culture supernatant contains growth factors and cytokines such as stem cell growth factors, fibroblast growth factors, and interleukins. Nebulizing both can deliver them directly to the lungs through the respiratory tract, achieving highly efficient local treatment. However, the application of the above-mentioned technologies relies on the efficient separation of high-purity exosomes and culture supernatants rich in active ingredients. Existing separation methods often suffer from problems such as low recovery rate, insufficient purity, and impaired activity of active ingredients, which restricts the further development of this treatment technology. Therefore, we need to upgrade and modify the existing technologies to overcome the existing problems and shortcomings. Utility Model Content

[0003] The purpose of this invention is to provide a dual-tangential flow microfluidic exosome separation device to solve the problems mentioned in the background art.

[0004] To achieve the above objectives, this utility model provides the following technical solution: Design a dual tangential flow microfluidic exosome separation device, including a centrifugation component, an ultrafiltration component, a liquid inlet pipeline and a collection component. The centrifugation component is connected to the ultrafiltration component through the liquid inlet pipeline, and the collection component is located at the liquid outlet end of the ultrafiltration component. The centrifugal assembly includes a centrifugal tank, a centrifugal motor, and a lower base. The centrifugal tank is installed at the output end of the centrifugal motor, and the centrifugal motor is fixed on the lower base. The collection assembly includes an exosome collection bottle, a supernatant collection bottle, and a diversion valve. The exosome collection bottle and the supernatant collection bottle are connected to the ultrafiltration assembly via the diversion valve.

[0005] Preferably, the ultrafiltration module is provided with a flow valve and a check valve at both ends, the flow valve is located at the inlet of the ultrafiltration module, and the check valve is located at the outlet of the ultrafiltration module.

[0006] Preferably, the centrifuge tank is equipped with a speed sensor and a time controller. The speed sensor is installed on the inner side wall of the centrifuge tank, and the time controller is electrically connected to the centrifuge motor.

[0007] Preferably, the inner wall of the centrifuge tank is provided with an anti-corrosion coating and the bottom is provided with a slag discharge port, and the anti-corrosion coating is made of polytetrafluoroethylene material.

[0008] Preferably, the diversion valve is a three-way valve, with its inlet connected to the outlet of the ultrafiltration component, and its two outlets connected to the exosome collection bottle and the supernatant collection bottle, respectively. The diversion valve is also electrically connected to the control box.

[0009] Preferably, both the exosome collection bottle and the supernatant collection bottle are made of transparent glass and are marked with graduation lines.

[0010] Preferably, the centrifugation assembly has a control box at its front end, which is electrically connected to the centrifugation assembly, the ultrafiltration assembly, and the collection assembly, respectively. The surface of the control box is equipped with a display screen and operation buttons.

[0011] Compared with the prior art, the beneficial effects of this utility model are as follows: 1. This utility model, through the inclusion of a centrifugation component, an ultrafiltration component, and a collection component within the device, achieves efficient and precise separation of exosomes by initially separating large particulate impurities in the culture system through the centrifugation component, separating the culture supernatant permeate and exosomes through the ultrafiltration membrane inside the ultrafiltration component, and collecting the culture supernatant permeate and exosomes separately through the collection component. This facilitates the reduction of loss of effective components during the separation process, improves the recovery rate and purity of exosomes, simplifies the exosome separation operation process, reduces errors caused by human intervention, enhances the repeatability and reliability of experiments, and allows for the simultaneous acquisition of high-purity exosomes and culture supernatant rich in active ingredients, meeting the needs of different research scenarios for both components.

[0012] Specific embodiments of the present invention are disclosed in detail with reference to the following description and accompanying drawings, indicating how the principles of the present invention can be employed. It should be understood that the embodiments of the present invention are not limited in scope. Within the spirit and scope of the appended claims, the embodiments of the present invention include many changes, modifications, and equivalents. Attached Figure Description

[0013] The accompanying drawings are provided to further illustrate the present invention and form part of the specification. They are used together with the embodiments of the present invention to explain the present invention, but do not constitute a limitation thereof. In the drawings: Figure 1 This is a schematic diagram of the overall structure according to the present utility model; Figure 2A schematic diagram of the centrifuge assembly according to this utility model; Figure 3 A schematic diagram of the ultrafiltration assembly according to this utility model; Figure 4 This is a schematic diagram of the collection components according to the present invention.

[0014] In the diagram: 1. Centrifuge assembly; 11. Centrifuge tank; 12. Centrifuge motor; 13. Lower base; 2. Ultrafiltration assembly; 21. Flow valve; 22. Check valve; 3. Inlet pipeline; 4. Collection assembly; 41. Exosome collection bottle; 42. Supernatant collection bottle; 43. Diverter valve; 5. Control box. Detailed Implementation

[0015] To make the technical means, creative features, objectives and effects of this utility model easier to understand, the present utility model will be further described below in conjunction with specific embodiments.

[0016] like Figure 1-4 As shown, this embodiment provides a dual tangential flow microfluidic exosome separation device, including a centrifugation component 1, an ultrafiltration component 2, an inlet pipe 3, and a collection component 4. The centrifugation component 1 is connected to the ultrafiltration component 2 through the inlet pipe 3. The collection component 4 is located at the outlet end of the ultrafiltration component 2. A control box 5 is provided at the front end of the centrifugation component 1. The control box 5 is electrically connected to the centrifugation component 1, the ultrafiltration component 2, and the collection component 4 respectively. The surface of the control box 5 is provided with a display screen and operation buttons. In this embodiment, the centrifugation assembly 1 includes a centrifuge tank 11, a centrifuge motor 12, and a lower base 13. The centrifuge tank 11 is installed at the output end of the centrifuge motor 12, and the centrifuge motor 12 is fixed on the lower base 13. The centrifuge tank 11 is equipped with a speed sensor and a time controller. The speed sensor is installed on the inner side wall of the centrifuge tank 11, and the time controller is electrically connected to the centrifuge motor 12. The inner wall of the centrifuge tank 11 is provided with an anti-corrosion coating, and the bottom is provided with a slag discharge port. The anti-corrosion coating is made of polytetrafluoroethylene material. The centrifuge motor 12 drives the centrifuge tank 11 to work on the lower base 13. The speed sensor and the time controller monitor the speed of the centrifuge tank 11 and control the working time. This can initially separate large particulate impurities in the culture system, which is conducive to laying the foundation for the subsequent ultrafiltration process, reducing the clogging of the ultrafiltration membrane by large particulate impurities, extending the service life of the ultrafiltration membrane, improving the efficiency and accuracy of subsequent ultrafiltration separation, and ensuring better separation effect of exosomes and culture supernatant. In this embodiment, the collection component 4 includes an exosome collection bottle 41, a supernatant collection bottle 42, and a diversion valve 43. The exosome collection bottle 41 and the supernatant collection bottle 42 are connected to the ultrafiltration component 2 via the diversion valve 43. The diversion valve 43 is a three-way valve, with its inlet connected to the outlet of the ultrafiltration component 2 and its two outlets connected to the exosome collection bottle 41 and the supernatant collection bottle 42, respectively. The diversion valve 43 is also electrically connected to the control box 5. Both the exosome collection bottle 41 and the supernatant collection bottle 42 are made of transparent glass with graduation lines marked on the bottle body. The separated exosomes and culture supernatant enter the exosome collection bottle 41 and the supernatant collection bottle 42 respectively through the diversion valve 43, which can achieve effective separation and collection of exosomes. It is convenient to intuitively grasp the collection volume of exosomes and culture supernatant through the transparent bottle body and graduation lines, and accurately control the separation process. It is also convenient for subsequent storage, detection, or further research of the two components, reducing interference caused by mixing of different components.

[0017] In this embodiment, the ultrafiltration component 2 is provided with a flow valve 21 and a one-way valve 22 at both ends. The flow valve 21 is located at the inlet of the ultrafiltration component 2, and the one-way valve 22 is located at the outlet of the ultrafiltration component 2. The culture supernatant permeates and exosomes are separated by the ultrafiltration membrane inside the ultrafiltration component 2. The flow valve 21 is used to regulate the flow rate of the liquid entering the ultrafiltration component 2, and the one-way valve 22 can prevent the liquid from flowing back from the ultrafiltration component 2 to the centrifuge component 1.

[0018] The working principle and process of this utility model are as follows: During operation, the operator sets parameters such as centrifugation speed, time, and the opening degree of flow valve 21 through the control box 5, and the parameters are displayed on the screen. The mesenchymal stem cell culture system is introduced into the centrifuge tank 11. After the device is started, the centrifuge motor 12 drives the centrifuge tank 11 to rotate at high speed on the lower base 13. The speed sensor monitors the speed and provides feedback, the time controller controls the duration, large particles of impurities settle to the bottom of the tank under the action of centrifugal force, the polytetrafluoroethylene coating protects the tank body, and after centrifugation, the supernatant containing exosomes flows to the ultrafiltration component 2 through the inlet pipe 3. The flow valve 21 adjusts the flow rate, and the liquid passes through the 30-200nm pore size ultrafiltration membrane. The culture supernatant permeates through the membrane, while the exosomes are retained. The one-way valve 22 prevents backflow and ensures stable separation. The separated exosome concentrate and culture supernatant enter the diversion valve 43 and, under the control of the control box 5, are respectively introduced into the exosome collection bottle 41 and the supernatant collection bottle 42. The transparent bottle and graduation lines make it easy to observe the collection volume. After completion, impurities are discharged through the slag outlet, achieving efficient and precise separation and ensuring the activity of effective ingredients.

[0019] In the description of this utility model, it should be noted that, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," "connected," etc., should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model according to the specific circumstances.

[0020] In the description of this embodiment, the terms "upper," "lower," "right," etc., refer to the orientation or positional relationship shown in the accompanying drawings. They are used only for ease of description and simplification of operation, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model. In addition, the terms "first" and "second" are only used for distinction in description and have no special meaning.

Claims

1. A dual-tangential flow microfluidic exosome separation device, characterized in that, It includes a centrifugal assembly (1), an ultrafiltration assembly (2), an inlet pipe (3), and a collection assembly (4). The centrifugal assembly (1) is connected to the ultrafiltration assembly (2) through the inlet pipe (3), and the collection assembly (4) is located at the outlet end of the ultrafiltration assembly (2). The centrifugal assembly (1) includes a centrifugal bucket (11), a centrifugal motor (12) and a lower base (13). The centrifugal bucket (11) is installed at the output end of the centrifugal motor (12), and the centrifugal motor (12) is fixed on the lower base (13). The collection component (4) includes an exosome collection bottle (41), a supernatant collection bottle (42), and a diversion valve (43). The exosome collection bottle (41) and the supernatant collection bottle (42) are connected to the ultrafiltration component (2) via the diversion valve (43).

2. The dual-tangential flow microfluidic exosome separation device according to claim 1, characterized in that: The ultrafiltration module (2) is provided with a flow valve (21) and a check valve (22) at both ends. The flow valve (21) is located at the inlet of the ultrafiltration module (2), and the check valve (22) is located at the outlet of the ultrafiltration module (2).

3. The dual-tangential flow microfluidic exosome separation device according to claim 1, characterized in that: The centrifuge tank (11) is equipped with a speed sensor and a time controller. The speed sensor is installed on the inner side wall of the centrifuge tank (11), and the time controller is electrically connected to the centrifuge motor (12).

4. The dual-tangential flow microfluidic exosome separation device according to claim 1, characterized in that: The centrifuge tank (11) has an anti-corrosion coating on its inner wall and a slag discharge port at the bottom. The anti-corrosion coating is made of polytetrafluoroethylene.

5. The dual-tangential flow microfluidic exosome separation device according to claim 1, characterized in that: The diversion valve (43) is a three-way valve. Its inlet is connected to the outlet of the ultrafiltration component (2), and its two outlets are connected to the exosome collection bottle (41) and the supernatant collection bottle (42) respectively. The diversion valve (43) is electrically connected to the control box (5).

6. The dual-tangential flow microfluidic exosome separation device according to claim 1, characterized in that: Both the exosome collection bottle (41) and the supernatant collection bottle (42) are made of transparent glass and are marked with graduation lines.

7. The dual-tangential flow microfluidic exosome separation device according to claim 1, characterized in that: The centrifugal assembly (1) is equipped with a control box (5) at the front end. The control box (5) is electrically connected to the centrifugal assembly (1), the ultrafiltration assembly (2) and the collection assembly (4) respectively. The surface of the control box (5) is equipped with a display screen and operation buttons.

Images