Low-temperature exosome extraction device and method for using same

Abstract

The present invention relates to the field of exosome extraction. Disclosed are a low-temperature exosome extraction device and a method for using same. The low-temperature exosome extraction device comprises an extraction stand and a bearing platform fixed on the extraction stand, and further comprises a plurality of filter cylinders mounted on the bearing platform at equal angular intervals, a collection cylinder circumferentially slidably mounted on the extraction stand, a vacuum tube connected to the collection cylinder, and liquid intake tubes fixed on the extraction stand and used for adding an auxiliary exosome extraction reagent into the filter cylinders, wherein a sliding block is slidably mounted on the side wall of the collection cylinder, a connecting ring for connecting and sealing the collection cylinder and a filter cylinder is fixed on the sliding block, and a steering support is fixedly connected to the collection cylinder; and the extraction stand is provided with a steering assembly for controlling the collection cylinder to rotate to the position under different filter cylinders. The low-temperature exosome extraction device simplifies the extraction structure, and, during multiple filtration for exosome extraction, agitates and disperses a cell fluid to be filtered, thereby preventing macromolecular exosomes from clogging a filter membrane, and improving the purity of extracted exosomes.

Description

A cryogenic extraction device for exosomes and its usage method Technical Field

[0001] This invention relates to the field of exosome extraction, specifically to a low-temperature exosome extraction device and its usage method. Background Technology

[0002] Exosome extraction is an important step in cell biology research, mainly used to isolate and purify exosomes from complex biological samples. When performing exosome extraction, the degradation and denaturation of the sample contents should be avoided as much as possible. All operations should be carried out at 4°C. The extraction temperature needs to be strictly controlled during the exosome extraction process.

[0003] Chinese patent CN218596389U discloses an exosome extraction device, including a housing and a first filter, a second filter, and a tangential flow filter located within the housing and connected in sequence. The housing is equipped with a cooling element for cooling the interior. A vacuum pump is installed on the second filter to generate negative pressure within both the second and first filters. A first peristaltic pump is installed between the second filter and the tangential flow filter. The pore sizes of the first, second, and tangential flow filters decrease sequentially. This exosome extraction device not only maintains a low-temperature environment throughout the extraction process, effectively inhibiting enzyme activity in the extraction system to prevent enzymes from hydrolyzing proteins into numerous small impurity molecules, but also eliminates the need for expensive ultracentrifugation equipment, significantly shortening the extraction and separation time compared to ultracentrifugation, thus offering higher efficiency.

[0004] Multiple filtration extraction of exosomes involves multiple filtration structures, pumps for liquid transfer, and piping structures. The overall extraction equipment is relatively large. After multiple filtrations, the amount of liquid isolated on the filter membrane gradually decreases, and some exosomes precipitate on the filter membrane. Exosomes cannot be completely transferred to different filtration structures, resulting in exosome loss during the transfer and filtration process. Furthermore, the exosomes precipitated on the filter membrane also significantly affect the filtration efficiency, making it difficult to filter out some small molecules, thus affecting the purity of the final filtered exosomes. Summary of the Invention

[0005] To address the aforementioned issues, a low-temperature exosome extraction device and its usage method are provided. The device simplifies the multiple filtration structure through a steering component and a liquid delivery component. Furthermore, as the collection cylinder rotates to different filter cylinders, an oscillation component disperses the cell fluid within the filter cylinders. This solves the problems of multiple filtration structures, pumps for liquid transfer, and piping structures involved in exosome extraction, which result in a large overall extraction device size. Additionally, after multiple filtrations, the amount of liquid remaining on the filter membrane gradually decreases, leading to some exosomes settling on the membrane. Exosomes cannot be completely transferred to different filtration structures, resulting in exosome loss during the transfer and filtration process. The remaining exosomes on the filter membrane also significantly affect the filtration efficiency, making it difficult to filter out some small molecules and thus impacting the purity of the final filtered exosomes.

[0006] To address the problems of existing technologies, this invention provides a low-temperature exosome extraction device, comprising an extraction frame and a support platform fixed on the extraction frame. The exosome extraction device further includes multiple filter cylinders mounted at equal angles on the support platform, a collection cylinder slidably mounted on the extraction frame, a vacuum tube connected to the collection cylinder, and a liquid inlet pipe fixed on the extraction frame for adding auxiliary exosome extraction reagents to the filter cylinders. Filter membranes with different pore sizes are installed in the multiple filter cylinders. A sliding block is slidably mounted on the side wall of the collection cylinder, and a connecting ring connecting the sealed collection cylinder and the filter cylinder is fixed on the sliding block. A steering frame is fixedly connected to the collection cylinder, and a connecting lifting rod is fixedly mounted on the steering frame. A sliding block is fixed to the lifting end of the connecting lifting rod. The extraction frame is provided with a steering assembly for controlling the rotation of the collection cylinder to different filter cylinders, and a liquid conveying assembly for controlling the liquid transfer in different filter cylinders is provided on the steering assembly.

[0007] Preferably, the steering assembly includes a drive motor fixed on the extraction frame, a drive rod fixedly connected to the output end of the drive motor, a lower toothed column fixed on the drive rod, an inner toothed groove rod slidably sleeved on the lower toothed column, the inner toothed groove rod being movably connected to the support platform, an upper toothed column rotatably mounted on the extraction frame, and the upper toothed column and the inner toothed groove rod being slidably engaged; a displacement lifting rod is fixedly mounted on the bogie, a lifting ring is fixed to the lifting end of the displacement lifting rod, and the lifting ring is rotatably assembled with the inner toothed groove rod.

[0008] Preferably, the liquid delivery assembly includes a support frame fixedly sleeved on the internal toothed rod, a pump body fixedly installed on the support frame, an absorption pipe for absorbing exosome liquid connected to the pump body, and a discharge pipe for discharging exosome liquid connected to the pump body. The absorption pipe and the discharge pipe correspond to the positions of two circumferentially adjacent filter cartridges, respectively. The outlet ends of the absorption pipe and the discharge pipe are coaxially aligned with the filter cartridges, and the length of the absorption pipe is greater than the length of the discharge pipe.

[0009] Preferably, the inlet pipe is equipped with an oscillation component to control the oscillation of the filter cylinder, thereby preventing the filter membrane from being blocked by cell fluid precipitates.

[0010] Preferably, the oscillation assembly includes an S-shaped groove formed on the liquid inlet pipe, a slip ring slidably connected to the S-shaped groove, and a positioning ring rotatably mounted on the slip ring; an oscillation rod is fixedly connected to the slip ring, and an oscillation hammer for vibrating and striking the inner wall of the filter cylinder is fixedly connected to the oscillation rod.

[0011] Preferably, the extraction frame is rotatably mounted with a drive gear fixedly connected to the upper gear column, and the extraction frame is rotatably connected with a plurality of driven gears meshing with the drive gear. The number of driven gears is the same as the number of filter cartridges. A sliding column is eccentrically connected to the driven gear. A guide slide rail is fixedly mounted on the extraction frame, and a guide slide is slidably mounted on the guide slide rail. The guide slide is slidably mounted with the sliding column. A push rod is rotatably connected to the guide slide, and the push rod is rotatably connected to the positioning ring.

[0012] Preferably, the extraction rack is fitted with a low-temperature control component for regulating the extraction temperature of exosomes; the low-temperature control component includes a heat insulation cover fitted on the extraction rack, a cooling pipe installed on the heat insulation cover, a condenser fixed in the heat insulation cover, a compressor installed at one end of the connection between the condenser and the cooling pipe, a throttling valve installed at the other end of the connection between the condenser and the cooling pipe, and a fan installed on the heat insulation cover to control the heat dissipation of the condenser.

[0013] Preferably, the cooling pipe is connected to a diversion cooling component that uniformly regulates the temperature of exosome extraction in the filter cartridge.

[0014] Preferably, the diversion cooling assembly includes a cooling ring coaxially sleeved outside the filter cylinder, a cooling branch pipe installed in the cooling ring, and an inlet pipe and an outlet pipe for controlling the inflow and outflow of coolant connected through the cooling branch pipe. The inlet pipe and the outlet pipe are connected through the cooling pipe via telescopic conduits. A fixing frame is fixedly installed between the inlet pipe, the outlet pipe and the cooling ring, and a positioning frame is fixedly assembled between the fixing frame and the positioning ring.

[0015] The present invention also includes a method of using an exosome low-temperature extraction device, comprising the above-described exosome low-temperature extraction device, and including the following steps:

[0016] S1. The simplified extraction equipment involves multiple filtrations. Cell fluid is introduced into a filter cartridge through the inlet pipe. After initial filtration, the valve of the filter cartridge is closed. The liquid is then transferred from the filter cartridge to an adjacent filter cartridge via the liquid delivery assembly. The collection cartridge is moved to the bottom of different filter cartridges by the steering assembly. Multiple filter cartridges and a set of liquid delivery assemblies can complete multiple filtrations of the cell fluid.

[0017] S2. Agitate the cell fluid to assist in the extraction of exosomes. If the exosome particles are large and precipitate, they are prone to clogging the filter membrane. Agitate the filter tube while rotating the collection tube to avoid the exosomes in the filter tube precipitating and clogging the filter membrane.

[0018] S3. Adjust the exosome extraction temperature. Precisely control the ambient temperature for exosome extraction by adjusting the temperature of the filter cartridge to avoid degradation of exosomes during extraction.

[0019] The advantages of this invention compared to the prior art are:

[0020] 1. The present invention simplifies the equipment by incorporating a steering component and a liquid transfer component, and enables multiple filtration processes of cell fluid. After one filtration process, the cell fluid can be transferred to another filter cartridge via the liquid transfer component. The steering component can rotate to switch the positions of the collection cartridge and the liquid transfer component, allowing the cell fluid to be filtered multiple times. During the transfer process, the valve structure below the filter cartridge can be closed, and reagents to assist in exosome extraction can be added to the filter cartridge through the inlet pipe to increase the liquid volume and reduce the loss of exosomes during continuous transfer.

[0021] 2. In this invention, an oscillation component is provided to generate oscillation in the filter cylinder during the driving process of the steering component. The steering component drives the collection cylinder to rotate circumferentially. The upper gear column in the steering component drives the driving gear to rotate. Under the action of gear meshing transmission, the driven gear is controlled to rotate. The driven gear pushes the guide slide to move laterally and reciprocally through the sliding column. The guide slide controls the positioning ring to move longitudinally through the push rod. During the longitudinal movement, the positioning ring controls the sliding ring to move up and down outside the liquid inlet pipe while reciprocating. The rotation of the positioning ring causes the oscillation rod and oscillating hammer to strike the filter cylinder, so that the cell fluid in the filter cylinder is oscillated and dispersed, avoiding the precipitation of exosomes in the cell fluid on the filter membrane, reducing the loss during exosome transport, and also preventing exosome precipitation and blockage on the filter membrane.

[0022] 3. The present invention is equipped with a diversion cooling component, which can uniformly and effectively regulate the temperature of the filter cartridge for exosome extraction. The coolant flowing in the cooling pipe flows into the cooling branch pipe through the telescopic conduit and the water inlet pipe. The cooling branch pipe is sleeved on the outside of the filter cartridge, which can adjust the temperature around the filter cartridge at close range. When the positioning ring in the oscillation component moves longitudinally, the positioning ring moves up and down synchronously through the positioning frame. The up and down reciprocating movement of the cooling ring can expand the range of temperature regulation of the filter cartridge by the cooling branch pipe, improve the temperature regulation effect, and quickly and effectively maintain the cell fluid in the filter cartridge at a low temperature for exosome extraction. Attached Figure Description

[0023] Figure 1 is a three-dimensional structural diagram of the extraction rack of a low-temperature exosome extraction device.

[0024] Figure 2 is a three-dimensional structural diagram of the collection tube of a low-temperature exosome extraction device.

[0025] Figure 3 is a schematic diagram of the three-dimensional structure of the support platform of a low-temperature exosome extraction device.

[0026] Figure 4 is a three-dimensional structural diagram of the filter cartridge of a low-temperature exosome extraction device.

[0027] Figure 5 is a three-dimensional structural diagram of the inlet pipe of a low-temperature exosome extraction device.

[0028] Figure 6 is a schematic diagram of the three-dimensional structure of the internal toothed groove rod of a low-temperature exosome extraction device.

[0029] Figure 7 is a schematic diagram of the three-dimensional structure of a vacuum tube in a low-temperature exosome extraction device.

[0030] Figure 8 is a schematic diagram of the three-dimensional structure of the filter membrane of a low-temperature exosome extraction device.

[0031] Figure 9 is a schematic diagram of the three-dimensional structure of the upper toothed column of a low-temperature exosome extraction device.

[0032] Figure 10 is a schematic diagram of the three-dimensional structure of the drive gear of a low-temperature exosome extraction device.

[0033] Figure 11 is a schematic diagram of the driven gear structure of a cryogenic exosome extraction device.

[0034] Figure 12 is a schematic diagram of the push rod structure of a low-temperature exosome extraction device.

[0035] Figure 13 is a schematic diagram of the three-dimensional structure of a slip ring in a low-temperature exosome extraction device.

[0036] Figure 14 is a schematic diagram of the three-dimensional structure of the guide carriage of a low-temperature exosome extraction device.

[0037] Figure 15 is a three-dimensional structural diagram of the cooling pipe of a low-temperature exosome extraction device.

[0038] Figure 16 is a schematic diagram of the three-dimensional structure of the cooling ring of a low-temperature exosome extraction device.

[0039] Figure 17 is a three-dimensional structural diagram of the cooling branch pipe of a low-temperature exosome extraction device.

[0040] The diagram is labeled as follows: 1. Extraction frame; 11. Support platform; 12. Filter cartridge; 121. Filter membrane; 13. Collection cartridge; 131. Sliding block; 132. Connecting ring; 133. Bogie; 134. Connecting lifting rod; 14. Vacuum tube; 15. Liquid inlet pipe; 2. Steering assembly; 21. Drive motor; 22. Drive rotating rod; 23. Lower toothed column; 24. Internal toothed rod; 241. Shifting lifting rod; 242. Lifting ring; 25. Upper toothed column; 3. Liquid delivery assembly; 31. Support frame; 32. Pump body; 33. Absorption pipe; 34. Drain pipe; 4. Vibration assembly; 41. S-shaped chute; 42. Slip ring; 43. Positioning ring; 431. Active... 432. Gear; 433. Driven gear; 434. Sliding column; 435. Guide slide rail; 436. Guide carriage; 437. Push rod; 44. Vibrating rod; 45. Vibrating hammer; 5. Low temperature control component; 51. Insulation cover; 52. Cooling pipe; 53. Condenser; 54. Compressor; 55. Throttling valve; 56. Fan; 6. Diverter cooling component; 61. Cooling ring; 62. Cooling branch pipe; 63. Inlet pipe; 64. Outlet pipe; 65. Fixing bracket; 651. Positioning bracket; 66. Telescopic conduit. Detailed Implementation

[0041] To further understand the features, technical means, and specific objectives and functions achieved by the present invention, the present invention will be described in further detail below with reference to the accompanying drawings and specific embodiments.

[0042] Referring to Figures 1-9, a low-temperature exosome extraction device includes an extraction frame 1 and a support platform 11 fixed on the extraction frame 1. The exosome extraction device also includes multiple filter cylinders 12 mounted at equal angles on the support platform 11, a collection cylinder 13 circumferentially slidably mounted on the extraction frame 1, a vacuum tube 14 connected to the collection cylinder 13, and an inlet pipe 15 fixed on the extraction frame 1 for adding auxiliary exosome extraction reagents to the filter cylinders 12. Filter membranes 121 with different pore sizes are installed in the multiple filter cylinders 12. A sliding block 131 is slidably mounted on the side wall of the collection cylinder 13, and a connecting sealing device is fixed on the sliding block 131. The collecting cylinder 13 is connected to the filter cylinder 12 by a connecting ring 132. A bogie 133 is fixedly connected to the collecting cylinder 13, and a connecting lifting rod 134 is fixedly installed on the bogie 133. A sliding block 131 is fixed to the lifting end of the connecting lifting rod 134. The extraction frame 1 is equipped with a steering component 2 to control the rotation of the collecting cylinder 13 to different filter cylinders 12. The steering component 2 is equipped with a liquid conveying component 3 to control the liquid transfer in different filter cylinders 12. The collecting cylinder 13 is equipped with a vacuum pump and a gas filling device connected to a vacuum tube 14 to regulate the gas pressure inside the collecting cylinder 13 in order to assist in the filtration and extraction of exosomes from cell fluid.

[0043] Cell fluid is fed into one of the filter cartridges 12 via the inlet pipe 15. The connecting lifting rod 134 moves the sliding block 131 upwards, causing the connecting ring 132 to move upwards synchronously. The connecting ring 132 connects the filter cartridge 12 and the collection cartridge 13. The air pressure in the collection cartridge 13 is adjusted via the vacuum tube 14. The valve below the filter cartridge 12 is opened, allowing small molecules to pass through the filter membrane 121 in the filter cartridge 12 and enter the collection cartridge 13 below. After filtration, the valve below the filter cartridge 12 is closed. Reagents for exosome extraction are added to the filter cartridge 12 via the inlet pipe 15, increasing the solution volume in the filter cartridge 12. This allows the exosomes isolated on the filter membrane 121 to mix better in the liquid. Then, the liquid transport component 3 controls the transfer of the isolated liquid in the filter cartridge 12 to the adjacent filter cartridge 12. The connecting lifting rod 134 controls the connecting ring 132 to move down and be stored in the collection cartridge 13. Then, the steering component 2 controls the collection cartridge 13 to rotate circumferentially, so that the collection cartridge 13 rotates to correspond with the filter cartridge 12 containing cell fluid. The previous cell fluid filtration steps are repeated to filter and extract exosomes from the cell fluid again. After multiple extractions, the purity of exosome extraction can be improved. The steering component 2 and the liquid transport component 3 can simplify the composition structure of the multiple filtration device and effectively extract exosomes from the cell fluid.

[0044] Referring to Figures 3-9, the steering assembly 2 includes a drive motor 21 fixed on the extraction frame 1. The output end of the drive motor 21 is fixedly connected to a drive rod 22. A lower toothed column 23 is fixed on the drive rod 22. An inner toothed groove rod 24 is engaged and slidably sleeved on the lower toothed column 23. The inner toothed groove rod 24 is movably connected to the support platform 11. An upper toothed column 25 is rotatably mounted on the extraction frame 1. The upper toothed column 25 and the inner toothed groove rod 24 are engaged and slidably connected. A displacement lifting rod 241 is fixedly mounted on the bogie 133. A lifting ring 242 is fixed at the lifting end of the displacement lifting rod 241. The lifting ring 242 and the inner toothed groove rod 24 are rotatably assembled.

[0045] The operating displacement lifting rod 241 controls the lifting ring 242 to drive the internal toothed rod 24 to move upward. The internal toothed rod 24 drives the liquid delivery assembly 3 to move upward away from the filter cartridge 12. The operating drive motor 21 controls the drive rotating rod 22 to rotate. The rotation of the drive rotating rod 22 drives the lower toothed column 23, the internal toothed rod 24 and the upper toothed column 25 to rotate synchronously. The drive rotating rod 22 drives the bogie 133 and the collection cartridge 13 to rotate. The collection cartridge 13 rotates circumferentially against the extraction frame 1, so that the collection cartridge 13 rotates circumferentially to the bottom of another filter cartridge 12. The rotation of the internal toothed rod 24 drives the liquid delivery assembly 3 to rotate circumferentially, so that the liquid delivery assembly 3 rotates to correspond to the other two sets of filter cartridges 12. Thus, the collection cartridge 13, the filter cartridge 12 and the liquid delivery assembly 3 are combined to form a new cell fluid filtration assembly, so as to effectively filter and separate exosomes.

[0046] Referring to Figures 3-9, the liquid delivery assembly 3 includes a support frame 31 fixedly sleeved on the internal toothed rod 24. A pump body 32 is fixedly installed on the support frame 31. An absorption tube 33 for absorbing exosome fluid is connected to the pump body 32. A discharge tube 34 for discharging exosome fluid is also connected to the pump body 32. The absorption tube 33 and the discharge tube 34 correspond to the positions of two circumferentially adjacent filter cylinders 12. The outlet ends of the absorption tube 33 and the discharge tube 34 are coaxially aligned with the filter cylinders 12. The length of the absorption tube 33 is greater than the length of the discharge tube 34. The filter membranes 121 in different filter cylinders 12 have different pore densities, which can filter cell fluids of different molecular sizes.

[0047] The pump body 32 absorbs the cell fluid in the filter cartridge 12 through the absorption pipe 33. The cell fluid enters another filter cartridge 12 through the drain pipe 34, so that the cell fluid filtered and isolated in one filter cartridge 12 can be transferred to a new filter cartridge 12 for further filtration and processing, thereby improving the purity of the extracted exosomes.

[0048] As shown in Figures 2-14, the inlet pipe 15 is equipped with an oscillation component 4 to control the oscillation of the filter cylinder 12, thereby preventing the filter membrane 121 from being blocked by cell fluid precipitates.

[0049] When the steering component 2 controls the rotation of the collection cylinder 13 and the liquid delivery component 3, it can drive the oscillation component 4 to oscillate and process the filter cylinder 12, disperse the cell fluid in the filter cylinder 12, and prevent the exosomes in the cell fluid from precipitating and clogging the filter membrane 121.

[0050] Referring to Figures 2-14, the oscillation assembly 4 includes an S-shaped groove 41 opened on the liquid inlet pipe 15, a slip ring 42 slidably connected to the S-shaped groove 41, and a positioning ring 43 rotatably mounted on the slip ring 42; an oscillation rod 44 is fixedly connected to the slip ring 42, and an oscillation hammer 45 for vibrating and striking the inner wall of the filter cylinder 12 is fixedly connected to the oscillation rod 44.

[0051] The control positioning ring 43 is sleeved on the outside of the liquid inlet pipe 15 and moves longitudinally back and forth. The positioning ring 43 drives the slip ring 42 to move up and down back and forth synchronously. The slip ring 42 moves longitudinally along the direction of the S-shaped sliding groove 41. While the slip ring 42 moves up and down, it rotates back and forth. The oscillating rod 44 and oscillating hammer 45 fixed on the slip ring 42 move longitudinally and rotate back and forth synchronously. The oscillating rod 44 and oscillating hammer 45 vibrate and strike the filter cylinder 12, causing the filter cylinder 12 to vibrate. This disperses the particulate matter in the cell fluid in the filter cylinder 12 and prevents large molecular exosomes from being filtered and separated on the filter membrane 121.

[0052] Referring to Figures 4-14, a drive gear 431 fixedly connected to the upper gear column 25 is rotatably mounted on the extraction frame 1. Multiple driven gears 432 meshing with the drive gear 431 are rotatably connected to the extraction frame 1. The number of driven gears 432 is the same as the number of filter cylinders 12. A sliding column 433 is eccentrically connected to the driven gear 432. A guide slide rail 434 is fixedly mounted on the extraction frame 1. A guide slide 435 is slidably mounted on the guide slide rail 434. The guide slide 435 and the sliding column 433 are slidably assembled. A push rod 436 is rotatably connected to the guide slide 435. The push rod 436 is rotatably connected to the positioning ring 43.

[0053] When the upper gear 25 in the steering assembly 2 rotates, it drives the driving gear 431 to rotate synchronously. The driven gear 432, which is meshed with the driving gear 431, rotates accordingly. The sliding column 433, which is connected to the driven gear 432 off-center, rotates in a circle. When the sliding column 433 rotates in a circle, it pushes the guide slide 435 to move back and forth along the guide slide rail 434. The two ends of the push rod 436, which is connected between the guide slide 435 and the positioning ring 43, rotate accordingly. The rotating push rod 436 pushes the positioning ring 43, which is sleeved on the outside of the liquid inlet pipe 15, to move longitudinally back and forth. This drives the sliding ring 42 to move longitudinally and rotate along the S-shaped slide groove 41, thereby driving the oscillating rod 44 and the oscillating hammer 45 to oscillate and strike the filter cylinder 12, dispersing the cell fluid inside.

[0054] Referring to Figures 1, 2, and 15-17, a low-temperature control component 5 for regulating the extraction temperature of exosomes is installed on the extraction rack 1. The low-temperature control component 5 includes a heat insulation cover 51 installed on the extraction rack 1, a cooling pipe 52 installed on the heat insulation cover 51, a condenser 53 fixed in the heat insulation cover 51, a compressor 54 installed at one end of the connection between the condenser 53 and the cooling pipe 52, and a throttling valve 55 installed at the other end of the connection between the condenser 53 and the cooling pipe 52. A fan 56 for controlling the heat dissipation of the condenser 53 is installed on the heat insulation cover 51. The heat insulation cover 51 is divided into two modules. One module is used to install components such as the condenser 53 and the cooling pipe 52, and it is fixedly installed on the outside of the extraction rack 1. The other module is slidably installed on the extraction rack 1, and can be opened to inspect the structure in the extraction rack 1. This module is provided with an inspection door, which can be opened to observe the structure in the extraction rack 1.

[0055] The compressor 54 controls the coolant to enter the condenser 53. During the flow of the coolant in the condenser 53, it is rapidly cooled by the action of the fan 56. The cooled coolant enters the cooling pipe 52 through the throttle valve 55. This cycle continues, allowing the low-temperature coolant to flow in the cooling pipe 52. The low temperature in the cooling pipe 52 is transferred to the extraction rack 1, keeping the exosome structure extracted in the extraction rack 1 at a low temperature, thus preventing the exosome from decomposing due to temperature during the extraction process.

[0056] Referring to Figures 15-17, a split cooling component 6 is connected to the cooling pipe 52 to uniformly regulate the temperature of exosome extraction in the filter cartridge 12; the split cooling component 6 can independently affect a single filter cartridge 12 and stably maintain the temperature of exosome extraction from cell fluid in the filter cartridge 12.

[0057] Part of the coolant circulating in the cooling pipe 52 flows into the diversion cooling assembly 6, which can further regulate the ambient temperature around the filter cartridge 12 in order to maintain a relatively stable temperature for the extraction of exosomes in the filter cartridge 12.

[0058] Referring to Figures 15-17, the diversion cooling assembly 6 includes a cooling ring 61 coaxially sleeved outside the filter cartridge 12. A cooling branch pipe 62 is installed in the cooling ring 61. An inlet pipe 63 and an outlet pipe 64 for controlling the inflow and outflow of coolant are connected through the cooling branch pipe 62. The inlet pipe 63 and the outlet pipe 64 are connected through the telescopic conduit 66 to the cooling pipe 52. A fixing bracket 65 is fixedly installed between the inlet pipe 63, the outlet pipe 64 and the cooling ring 61. A positioning bracket 651 is fixedly assembled between the fixing bracket 65 and the positioning ring 43.

[0059] The coolant circulating in the cooling pipe 52 partially enters the inlet pipe 63 through the telescopic conduit 66. The coolant in the inlet pipe 63 flows into the cooling branch pipe 62, which is sleeved on the outside of the filter cartridge 12 to regulate the ambient temperature around the filter cartridge 12. The coolant flowing in the cooling branch pipe 62 eventually re-enters the cooling pipe 52 through the outlet pipe 64 and the telescopic conduit 66, maintaining the circulation of coolant in the low-temperature control component 5. When the positioning ring 43 in the oscillation component 4 moves longitudinally back and forth, the positioning ring 43 drives the fixing frame 65 to move longitudinally synchronously through the positioning bracket 651. The fixing frame 65 is fixed on the outside of the cooling ring 61, controlling the up-and-down reciprocating movement of the cooling ring 61 and the cooling branch pipe 62, expanding the range of ambient temperature regulation of the filter cartridge 12 by the cooling branch pipe 62. When the fixing frame 65 moves longitudinally back and forth, the telescopic conduit 66 extends and retracts autonomously to maintain the precise up-and-down movement of the cooling ring 61 to regulate the temperature of the filter cartridge 12.

[0060] Referring to Figures 1-17, the present invention also includes a method of using an exosome low-temperature extraction device, comprising the following steps:

[0061] S1. The simplified extraction equipment involves multiple filtrations. Cell fluid is introduced into a filter cartridge 12 through the inlet pipe 15. After preliminary filtration, the valve of the filter cartridge 12 is closed. The liquid in the filter cartridge 12 is introduced into an adjacent filter cartridge 12 through the liquid delivery assembly 3. The collection cartridge 13 is moved to the bottom of different filter cartridges 12 by the steering assembly 2. Multiple filter cartridges 12 and a set of liquid delivery assemblies 3 can complete multiple filtrations of cell fluid.

[0062] S2. Agitation of cell fluid assists in the extraction of exosomes. If the exosome particles are large and precipitate, they are prone to clogging the filter membrane 121. Agitation of the filter tube 12 is performed while rotating the collection tube 13 to prevent the exosomes in the filter tube 12 from precipitating and clogging the filter membrane 121.

[0063] S3. Adjust the exosome extraction temperature. The ambient temperature for exosome extraction is precisely controlled by adjusting the temperature of the filter cartridge 12 to avoid degradation of exosomes during extraction.

[0064] The above embodiments only illustrate one or more implementations of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of the present invention should be determined by the appended claims.

Claims

1. A cryogenic extraction device for exosomes, comprising an extraction frame (1) and a support platform (11) fixed on the extraction frame (1), characterized in that: The exosome extraction device also includes multiple filter cylinders (12) installed at equal angles on the support platform (11), a collection cylinder (13) circumferentially slidably installed on the extraction frame (1), a vacuum tube (14) connected to the collection cylinder (13), and an inlet tube (15) fixed on the extraction frame (1) for adding auxiliary exosome extraction reagents into the filter cylinder (12); Multiple filter cartridges (12) are equipped with filter membranes (121) with different pore sizes; A sliding block (131) is slidably installed on the side wall of the collection cylinder (13). A connecting ring (132) connecting the sealed collection cylinder (13) and the filter cylinder (12) is fixed on the sliding block (131). A bogie (133) is fixedly connected to the collection cylinder (13). A connecting lifting rod (134) is fixedly installed on the bogie (133). The sliding block (131) is fixed at the lifting end of the connecting lifting rod (134). The extraction rack (1) is provided with a steering assembly (2) for controlling the rotation of the collection cylinder (13) to different filter cylinders (12), and the steering assembly (2) is provided with a liquid conveying assembly (3) for controlling the liquid transfer in different filter cylinders (12).

2. The exosome low-temperature extraction device according to claim 1, characterized in that: The steering assembly (2) includes a drive motor (21) fixed on the extraction frame (1), a drive rod (22) fixedly connected to the output end of the drive motor (21), a lower toothed column (23) fixed on the drive rod (22), an inner toothed rod (24) engaged and slidably sleeved on the lower toothed column (23), the inner toothed rod (24) being movably connected to the support platform (11), and an upper toothed column (25) rotatably mounted on the extraction frame (1), the upper toothed column (25) and the inner toothed rod (24) being engaged and slidably connected; A displacement lifting rod (241) is fixedly installed on the bogie (133). A lifting ring (242) is fixed at the lifting end of the displacement lifting rod (241). The lifting ring (242) is rotatably assembled with the internal toothed rod (24).

3. The exosome low-temperature extraction device according to claim 2, characterized in that: The liquid delivery assembly (3) includes a support frame (31) fixedly sleeved on the inner toothed rod (24), a pump body (32) fixedly installed on the support frame (31), an absorption pipe (33) for absorbing exosome liquid connected to the pump body (32), and a discharge pipe (34) for discharging exosome liquid connected to the pump body (32). The absorption pipe (33) and the discharge pipe (34) correspond to the positions of two adjacent filter cylinders (12) on the circumference. The outlet ends of the absorption tube (33) and the drain tube (34) are coaxially aligned with the filter cylinder (12), and the length of the absorption tube (33) is greater than the length of the drain tube (34).

4. The exosome low-temperature extraction device according to claim 1, characterized in that: The inlet pipe (15) is equipped with an oscillation component (4) to control the oscillation of the filter cylinder (12) so as to prevent the filter membrane (121) from being blocked by cell fluid precipitates.

5. The exosome low-temperature extraction device according to claim 4, characterized in that: The oscillation assembly (4) includes an S-shaped groove (41) opened on the liquid inlet pipe (15), a slip ring (42) is slidably connected on the S-shaped groove (41), and a positioning ring (43) is rotatably mounted on the slip ring (42). An oscillating rod (44) is fixedly connected to the slip ring (42), and an oscillating hammer (45) for vibrating and striking the inner wall of the filter cylinder (12) is fixedly connected to the oscillating rod (44).

6. The exosome low-temperature extraction device according to claim 5, characterized in that: The extraction frame (1) is rotatably mounted with a drive gear (431) fixedly connected to the upper gear column (25). The extraction frame (1) is rotatably connected with a plurality of driven gears (432) meshing with the drive gear (431). The number of driven gears (432) is the same as the number of filter cylinders (12). The driven gears (432) are eccentrically connected with a sliding column (433). The extraction frame (1) is fixedly mounted with a guide slide rail (434). The guide slide rail (434) is slidably mounted with a guide slide (435). The guide slide (435) is slidably mounted with the sliding column (433). The guide slide (435) is rotatably connected with a push rod (436). The push rod (436) is rotatably connected with a positioning ring (43).

7. The exosome low-temperature extraction device according to claim 1, characterized in that: The extraction rack (1) is fitted with a low-temperature control component (5) for regulating the temperature of exosome extraction; The low-temperature control component (5) includes a heat insulation cover (51) fitted on the extraction rack (1), a cooling pipe (52) installed on the heat insulation cover (51), a condenser (53) fixed in the heat insulation cover (51), a compressor (54) installed at one end of the connection between the condenser (53) and the cooling pipe (52), a throttle valve (55) installed at the other end of the connection between the condenser (53) and the cooling pipe (52), and a fan (56) installed on the heat insulation cover (51) to control the heat dissipation of the condenser (53).

8. The exosome low-temperature extraction device according to claim 7, characterized in that: The cooling pipe (51) is connected to a diversion cooling assembly (6) that uniformly regulates the temperature of exosome extraction in the filter cartridge (12).

9. The exosome low-temperature extraction device according to claim 8, characterized in that: The diversion cooling assembly (6) includes a cooling ring (61) coaxially sleeved outside the filter cylinder (12), a cooling branch pipe (62) is installed in the cooling ring (61), and an inlet pipe (63) and an outlet pipe (64) for controlling the inflow and outflow of coolant are connected through the cooling branch pipe (62). The inlet pipe (63) and the outlet pipe (64) are connected through the cooling pipe (52) via a telescopic conduit (66). A fixing bracket (65) is fixedly installed between the water inlet pipe (63), the water outlet pipe (64) and the cooling ring (61), and a positioning bracket (651) is fixedly assembled between the fixing bracket (65) and the positioning ring (43).

10. A method of using a low-temperature exosome extraction device, characterized in that, The exosome low-temperature extraction apparatus according to any one of claims 1-9 comprises the following steps: S1. The simplified extraction equipment filters multiple times. The cell fluid is introduced into a filter tube (12) through the liquid inlet pipe (15). After preliminary filtration, the valve of the filter tube (12) is closed. The liquid in the filter tube (12) is introduced into the adjacent filter tube (12) through the liquid delivery component (3). The collection tube (13) is moved to the bottom of different filter tubes (12) through the steering component (2). Multiple filter tubes (12) and a set of liquid delivery components (3) can complete multiple filtrations of the cell fluid. S2. Shaking cell fluid to assist in exosome extraction. If the exosome particles are large and precipitate, they are easy to clog the filter membrane (121). Shaking the filter tube (12) during the rotation of the collection tube (13) can prevent the exosomes in the filter tube (12) from precipitating and clogging the filter membrane (121). S3. Adjust the exosome extraction temperature. The ambient temperature for exosome extraction is precisely controlled by adjusting the temperature of the filter cartridge (12) to avoid degradation of exosomes during extraction.

Images