An exosome purification device

Abstract

This invention discloses an exosome purification device, relating to the field of stem cell processing technology. The device includes a centrifugation mechanism comprising an outer shell containing a rotatable centrifuge cylinder, with a detachable cap on the top of the shell; and a collection bottle connected to a first pipe at its top, which freely penetrates the cap and is positioned at the bottom of the centrifuge cylinder. A second pipe is also connected to the collection bottle, and this second pipe is connected to a negative pressure generating mechanism. Using this device effectively simplifies the separation and purification steps, minimizes contact with the supernatant during separation, and all operations are performed within the centrifuge cylinder, effectively avoiding cross-contamination of the sample solution and preserving the integrity of the exosomes.

Description

Technical Field

[0001] This utility model relates to the field of stem cell processing technology, and in particular to an exosome purification device. Background Technology

[0002] Exosomes, as important mediators of intercellular communication, have significant research and application value in biomedicine and other fields, and their separation and purification techniques are fundamental to research. Currently, the commonly used separation method is ultracentrifugation. Although this method is time-consuming, it yields exosomes with high purity.

[0003] Therefore, several devices based on the principle of rotary centrifugation have emerged in the market. For example, an exosome purification device with patent application number 202222961688.2 first uses a filter assembly to perform preliminary filtration of the sample solution, and then uses a motor to drive the centrifuge tank to rotate for centrifugal purification. Generally, a single centrifugation is insufficient to obtain high-purity exosomes, requiring multiple centrifugation operations to improve purity. Using the above-mentioned device, if secondary or multiple centrifugation purification is performed, all the supernatant and precipitate after the initial centrifugation in the centrifuge tank must be removed, and then the supernatant must be reinjected into the centrifuge tank separately. This process is not only cumbersome and prolongs the experimental cycle, but also increases the risk of cross-contamination of exosome samples during repeated transfer and sample switching. Utility Model Content

[0004] This invention aims to at least partially solve one of the technical problems in related technologies. To this end, this invention proposes an exosome purification device.

[0005] The technical solution to the technical problem solved by this utility model is as follows:

[0006] This utility model proposes an exosome purification device, including a centrifugal separation mechanism, comprising an outer shell, a drive motor disposed within the outer shell, a centrifuge cylinder whose output shaft is drively connected to the centrifuge cylinder, the centrifuge cylinder being detachably placed within the outer shell, and a cap being detachably connected to the top of the outer shell; a collection bottle, with a first pipe connected to its top, the first pipe freely penetrating the cap, and the suction end of the first pipe being placed at the bottom of the centrifuge cylinder; a second pipe also connected to the top of the collection bottle, the second pipe being connected to a negative pressure generating mechanism.

[0007] Preferably, the outer shell is provided with a rotatable drive cylinder via a bearing, the bottom of the drive cylinder is connected to the output shaft of the drive motor, and the centrifuge cylinder is detachably inserted into the drive cylinder.

[0008] Preferably, the bottom of the drive cylinder has a funnel-shaped groove, and the bottom of the centrifuge cylinder is conical. When the centrifuge cylinder is inserted into the drive cylinder, the conical bottom fits into the groove.

[0009] Preferably, the inner sidewall and bottom wall of the drive cylinder are provided with at least two sets of slots, and the outer sidewall and bottom surface of the centrifuge cylinder are fixedly connected with limit strips. The limit strips are adapted to be inserted into the slots so that the drive cylinder drives the centrifuge cylinder to rotate synchronously.

[0010] Preferably, a first cylinder edge and a second cylinder edge are respectively provided above the drive cylinder and the centrifuge cylinder, the second cylinder edge overlaps the first cylinder edge, and the maximum diameter of the second cylinder edge is smaller than the maximum diameter of the first cylinder edge.

[0011] Preferably, a pressure plug is provided inside the cap via a rotating shaft. After the cap is connected to the top of the outer shell, the pressure plug can press and seal the top of the centrifuge cylinder.

[0012] Preferably, a perforation is provided at the center of the cap and the plug, and a sealing element is provided to seal the perforation; the first pipe suction end can be placed through the perforation at the bottom of the centrifuge.

[0013] Preferably, a filter disc is detachably mounted on the top of the centrifuge cylinder, with the edge of the filter disc overlapping the top wall of the centrifuge cylinder.

[0014] Preferably, it also includes a body, which has a centrifugation chamber for placing a centrifugation separation mechanism, a collection chamber for placing a collection bottle, and a negative pressure chamber for placing a negative pressure generating mechanism.

[0015] Preferably, an operating platform is provided on the top of the machine body, and the operating platform is controlled and connected to the drive motor and the negative pressure generating mechanism.

[0016] The above technical solution has the following advantages or beneficial effects:

[0017] 1. In this invention, the sample solution is centrifuged using a centrifuge tube. After initial centrifugation, the precipitate at the bottom can be extracted by the cooperation of a negative pressure generating mechanism and a collection bottle, and then the centrifugation operation can be repeated. In the above operation process, the separation and purification steps can be effectively simplified, the contact with the supernatant is reduced, and all operations are carried out inside the centrifuge tube, which can effectively avoid cross-contamination of the sample solution and help maintain the integrity of the exosomes.

[0018] 2. In this utility model, the drive cylinder and the centrifuge cylinder are detachably connected through slots and limiting strips, which can provide high-strength torque transmission and thus avoid slippage and other phenomena. This plug-in connection can be disassembled simply by pulling the centrifuge cylinder upwards, which is suitable for scenarios involving large-scale centrifugation and frequent cleaning and replacement of centrifuge cylinders. Attached Figure Description

[0019] The accompanying drawings are provided to further understand 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 and do not constitute a limitation thereof.

[0020] Figure 1 This is a three-dimensional structural diagram of the sediment extraction process of this utility model;

[0021] Figure 2 This is an internal cross-sectional view of the sediment extraction process of this utility model;

[0022] Figure 3 A three-dimensional structural diagram of the present invention with added sample solution;

[0023] Figure 4 An internal cross-sectional view of the present invention with added sample solution filtration;

[0024] Figure 5 A three-dimensional structural diagram of the outer shell, centrifuge cylinder, cap, and drive cylinder;

[0025] Explanation of reference numerals in the attached figures:

[0026] 1. Main body; 2. Control panel;

[0027] 3. Centrifugal separation mechanism; 31. Outer shell; 32. Centrifuge cylinder; 33. Cap; 34. Drive cylinder; 35. Slot; 36. Limiting strip; 37. Plug; 38. Perforation; 39. Sealing component; 310. Drive motor;

[0028] 4. Collection bottle; 5. Negative pressure generating mechanism; 6. First pipeline; 8. Filter plate. Detailed Implementation

[0029] To make the objectives, features, and advantages of this utility model more apparent and understandable, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the embodiments described below are only some embodiments of this utility model, and not all embodiments. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this utility model.

[0030] It should be noted that in the description of this utility model, the terms "upper", "lower", "left", "right", "inner", "outer", etc., indicating the direction or positional relationship are based on the direction or positional relationship shown in the drawings. This is only for the convenience of description and does not indicate or imply that the device or element must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation of this utility model.

[0031] Furthermore, it should be noted that, in the description of this utility model, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of 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.

[0032] like Figure 1 as well as Figure 2 As shown, this embodiment proposes an exosome separation and purification device, which includes a centrifugal separation mechanism 3. The centrifugal separation mechanism 3 includes an outer shell 31, within which a drive motor 310 is installed. The output shaft of the drive motor 310 is connected to a centrifuge cylinder 32. The centrifuge cylinder 32 is used to load sample solutions and is driven to rotate at high speed by the drive motor 310 to achieve exosome separation and purification. The centrifuge cylinder 32 is detachably placed inside the outer shell 31. A cap 33 is detachably connected to the top of the outer shell 31. After the cap 33 closes the outer shell 31, the centrifuge cylinder 32 forms a closed cavity, ensuring the sealing and safety of the centrifugation process. It also includes a collection bottle 4, with a first pipe 6 connected to the top of the collection bottle 4. The first pipe 6 can freely pass through the cap 33, and its suction end is positioned at the bottom of the centrifuge cylinder 32. A second pipe is also connected to the top of the collection bottle 4, and the second pipe is connected to a negative pressure generating mechanism 5.

[0033] Using this design, during the separation and purification process, the sample solution is first added into the centrifuge tube 32, and the cap 33 is tightened to seal the outer shell 31, creating a closed space inside the centrifuge tube 32. This effectively prevents sample solution splashing during centrifugation, avoiding sample loss and environmental contamination. Subsequently, the drive motor 310 is started to rotate the centrifuge tube 32, completing the separation and purification of the sample solution.

[0034] After initial centrifugation, the sample solution will exhibit distinct stratification, consisting of a supernatant containing exosomes and a precipitate containing larger impurities (such as cell debris, dead cells, and large vesicles). The current conventional procedure involves transferring the exosome-containing supernatant into a new centrifuge tube 32 and centrifuging again. This process is cumbersome and carries the risk of contaminating the supernatant. This device employs a method that prioritizes the removal of the precipitate, extracting it from centrifuge tube 32 and retaining only the exosome-containing supernatant inside, facilitating subsequent centrifugation.

[0035] Specifically, the first pipe 6 is inserted through the cap 33 into the bottom of the centrifuge cylinder 32. The negative pressure generating mechanism 5 is activated, creating a negative pressure suction force inside the collection bottle 4. Under this suction force, the precipitate located at the bottom of the centrifuge cylinder 32 is drawn out through the first pipe 6 until only the supernatant containing exosomes remains in the centrifuge cylinder 32. At this point, the negative pressure generating mechanism 5 is turned off, the first pipe 6 is removed, and the drive motor 310 is activated to rotate the centrifuge cylinder 32 again, separating and purifying the exosome supernatant. By repeating the above steps and centrifuging multiple times, high-purity exosomes can be obtained.

[0036] This design utilizes centrifuge tube 32 for operation, avoiding the cumbersome process of "pouring, changing tubes, and centrifuging again." Multiple centrifugations can be completed within the same centrifuge tube 32, making it particularly suitable for batch extraction of exosomes and reducing manual intervention time. Furthermore, this design minimizes contact with the supernatant during operation, and since all operations take place within the centrifuge tube 32, it effectively prevents cross-contamination of the sample solution, thus preserving the integrity and biological activity of the exosomes.

[0037] In some embodiments, in order to achieve a structure that is both detachable and rotatable for the centrifuge cylinder 32, a drive cylinder 34 is provided inside the outer shell 31. The drive cylinder 34 is rotatably disposed inside the drive cylinder 34 via a bearing. The drive motor 310 is placed at the bottom of the drive cylinder 34 and connected to the bottom of the drive cylinder 34 to drive the drive cylinder 34 to rotate. The centrifuge cylinder 32 can then be detachably inserted into the drive cylinder 34 and rotate synchronously with the drive cylinder 34.

[0038] Further reference Figure 3 The drive cylinder 34 has at least two sets of slots 35 on its inner and bottom walls. Limiting strips 36 are fixedly connected to the outer side and bottom surface of the centrifuge cylinder 32. The limiting strips 36 are inserted into the slots 35 to allow the drive cylinder 34 to drive the centrifuge cylinder 32 to rotate synchronously. The insertion and engagement of the slots 35 and the limiting strips 36 provides high-strength torque transmission, ensuring that the centrifuge cylinder 32 rotates synchronously when the drive cylinder 34 rotates, thus avoiding slippage or lag. When the limiting strips 36 are inserted into the slots 35, they not only restrict the circumferential rotation of the centrifuge cylinder 32 but also ensure that the centrifuge cylinder 32 is vertically positioned within the drive cylinder 34 through axial limiting, preventing dynamic imbalance caused by axial movement during high-speed rotation.

[0039] After frequent centrifugation and purification operations, some precipitates in the sample solution inevitably adhere to the inner wall of the centrifuge tube 32, requiring cleaning of the centrifuge tube 32. This design adopts a plug-in installation method, and the centrifuge tube 32 can be removed simply by pulling it upwards, without the need for tools such as wrenches and screwdrivers. It is especially suitable for scenarios involving large-scale centrifugation and frequent cleaning and replacement of the centrifuge tube 32.

[0040] In some embodiments, such as Figure 3 as well as Figure 5 As shown, the bottom of the drive cylinder 34 has a funnel-shaped groove, and the bottom of the centrifuge cylinder 32 is conical. When the centrifuge cylinder 32 is inserted into the drive cylinder 34, the conical bottom fits into the groove. The conical bottom design inside the centrifuge cylinder 32 forms a gradually contracting structure, which allows the precipitate to gather towards the lowest point under centrifugal force. At the same time, it greatly reduces the possible dead corners, concentrates the precipitate in the same area, and makes it easier for the suction port of the first pipe 6 to accurately align with the gathering point, greatly improving the efficiency of precipitate suction.

[0041] In some embodiments, a first cylinder edge and a second cylinder edge are respectively provided above the drive cylinder 34 and the centrifuge cylinder 32. The second cylinder edge overlaps the first cylinder edge, and the maximum diameter of the second cylinder edge is smaller than the maximum diameter of the first cylinder edge. As shown in Figure 3, this design creates an operable space between the second cylinder edge and the inner wall of the outer casing 31, making it easy to grip the edge of the second cylinder edge with fingers and apply force to easily remove the centrifuge cylinder 32 from the outer casing 31.

[0042] Considering that the conventional connection between the cap 33 and the fixed pressure plug 37 during centrifugation may lead to severe wear of the pressure plug 37, this embodiment optimizes its structure to extend its service life: the pressure plug 37 is installed inside the cap 33 via a bearing, and the pressure plug 37 is made of rubber. When the cap 33 is connected to the top of the outer casing 31, the pressure plug 37 can press and seal the top of the centrifuge cylinder 32. The bearing enables relative rotation between the pressure plug 37 and the cap 33, preventing wear caused by rigid friction between the pressure plug 37 and the top of the centrifuge cylinder 32 during centrifugal rotation. The pressure plug 37 can also apply downward pressure to the centrifuge cylinder 32, causing the centrifuge cylinder 32 to rotate between the pressure plug 37 and the drive cylinder 34.

[0043] Furthermore, a perforation 38 is provided at the center of the cap 33 and the plug 37, and a sealing element 39 is provided for sealing the perforation 38. The suction end of the first pipe 6 can extend to the bottom of the centrifuge cylinder 32 through the perforation 38. The sealing element 39 can be a plug, nut, or other components. Considering that conventional plugs may be thrown out by centrifugal force during centrifugation, this embodiment adopts a combination structure of a threaded cylinder and a nut: the threaded cylinder is fixed above the perforation 38, and the nut is threadedly connected to the threaded cylinder. By tightening the nut, the perforation 38 is sealed, ensuring the stability and sealing performance of the sealing element 39 during centrifugation.

[0044] To ensure the accuracy of suction in the first pipe 6, a miniature camera is installed at the suction end of the first pipe 6 and electrically connected to the operating table 2.

[0045] In some embodiments, a filter disc 8 is detachably mounted above the centrifuge tube 32, with its edge overlapping the second edge of the top wall of the centrifuge tube 32. The filter disc 8 is designed to perform preliminary filtration of the solution when sample solution is injected into the centrifuge tube 32, intercepting larger cells. After the sample solution has been added, the filter disc 8 can be removed from above the centrifuge tube 32 by grasping its edge with fingers.

[0046] In some embodiments, the device further includes a body 1, which has a centrifugation chamber for housing the centrifugal separation mechanism 3, a collection chamber for housing the collection bottle 4, and a negative pressure chamber for housing the negative pressure generating mechanism 5. The device is controlled by an operating platform 2 located on top of the body 1, which is connected to the drive motor 310 and the negative pressure generating mechanism 5.

[0047] Furthermore, to improve the accuracy of suction in the first pipe 6, a miniature camera is installed at its suction end and electrically connected to the control panel 2. During suction, the operator can observe the imaging screen in real time through the control panel 2 and precisely control the position of the suction end of the first pipe 6, thereby significantly improving the suction effect of the precipitate.

[0048] In this design, the aforementioned components are integrated through the main body 1, which effectively reduces the space occupied and allows for direct transport as a whole, making it particularly suitable for inter-laboratory transfers. Furthermore, all components are detachable, facilitating replacement and maintenance.

[0049] Although the specific embodiments of the utility model have been described above in conjunction with the accompanying drawings, this is not intended to limit the scope of protection of the utility model. Based on the technical solution of the utility model, various modifications or variations that can be made by those skilled in the art without creative effort are still within the scope of protection of the utility model.

Claims

1. An exosome purification device, characterized in that, include: The centrifugal separation mechanism (3) includes an outer shell (31), a drive motor (310) is provided inside the outer shell (31), the output shaft of the drive motor (310) is connected to a centrifugal cylinder (32), the centrifugal cylinder (32) is detachably placed inside the outer shell (31), and a cap (33) is detachably connected to the top of the outer shell (31). The collection bottle (4) is connected to a first pipe (6) at the top. The first pipe (6) can freely pass through the cap (33) and the suction end of the first pipe (6) is placed at the bottom of the centrifuge cylinder (32). The collection bottle (4) is also connected to a second pipe at the top, and the second pipe is connected to a negative pressure generating mechanism (5).

2. The exosome purification apparatus according to claim 1, characterized in that, The outer shell (31) is provided with a rotatable drive cylinder (34) via a bearing. The bottom of the drive cylinder (34) is connected to the output shaft of the drive motor (310). The centrifuge cylinder (32) is detachably inserted into the drive cylinder (34).

3. The exosome purification apparatus according to claim 2, characterized in that, The bottom of the drive cylinder (34) is provided with a funnel-shaped groove, and the bottom of the centrifuge cylinder (32) is a conical bottom. When the centrifuge cylinder (32) is inserted into the drive cylinder (34), the conical bottom fits into the groove.

4. The exosome purification apparatus according to claim 2, characterized in that, At least two sets of slots (35) are provided on the inner side wall and bottom wall of the drive cylinder (34). Limiting strips (36) are fixedly connected to the outer side wall and bottom surface of the centrifuge cylinder (32). The limiting strips (36) are adapted to be inserted into the slots (35) so that the drive cylinder (34) drives the centrifuge cylinder (32) to rotate synchronously.

5. The exosome purification apparatus according to claim 2, characterized in that, The drive cylinder (34) and the centrifugal cylinder (32) are respectively provided with a first cylinder edge and a second cylinder edge. The second cylinder edge overlaps the first cylinder edge, and the maximum diameter of the second cylinder edge is smaller than the maximum diameter of the first cylinder edge.

6. The exosome purification apparatus according to claim 1, characterized in that, Inside the cap (33), a pressure plug (37) is provided via a rotating shaft. After the cap (33) is connected to the top of the outer shell (31), the pressure plug (37) can press and seal the top of the centrifuge tube (32).

7. The exosome purification apparatus according to claim 6, characterized in that, The cap (33) and the plug (37) are provided with a perforation (38) at the center and are equipped with a plugging part (39) to block the perforation (38); the suction end of the first pipe (6) can pass through the perforation (38) and be placed at the bottom of the centrifuge (32).

8. The exosome purification apparatus according to claim 1, characterized in that, A filter disc (8) is detachably mounted on the top of the centrifuge tube (32), with the edge of the filter disc (8) overlapping the top wall of the centrifuge tube (32).

9. The exosome purification apparatus according to claim 1, characterized in that, It also includes a body (1), which has a centrifugal chamber for placing the centrifugal separation mechanism (3), a collection chamber for placing the collection bottle (4), and a negative pressure chamber for placing the negative pressure generating mechanism (5).

10. An exosome purification apparatus according to claim 9, characterized in that, An operating table (2) is provided on the top of the machine body (1), and the operating table (2) is connected to the drive motor (310) and the negative pressure generating mechanism (5) for control.

Images