Exosome low-temperature extraction device and use method thereof

Abstract

The invention relates to the field of exosome extraction, and discloses an exosome low-temperature extraction device and a use method thereof.The exosome low-temperature extraction device comprises an extraction frame and a bearing table fixed to the extraction frame; the exosome extraction device further comprises a plurality of filter cartridges installed on the bearing table at equal angles, a collecting cartridge installed on the extraction frame in a circumferential sliding mode, a vacuum tube connected to the collecting cartridge, and a liquid inlet tube fixed in the filter cartridges and used for adding an auxiliary exosome extraction reagent. A sliding block is installed on the side wall of the collecting cylinder in a sliding mode, a connecting ring for connecting and sealing the collecting cylinder and the filter cylinder is fixed to the sliding block, and a steering frame is fixedly connected to the collecting cylinder; the extracting frame is provided with a steering assembly for controlling the collecting cylinder to rotate to the lower part of different filtering cylinders; according to the exosome low-temperature extraction device, the extraction structure is simplified, cell sap to be filtered is oscillated and dispersed when exosomes are filtered and extracted for multiple times, macromolecular exosomes are prevented from being blocked on a filter membrane, and the purity of the extracted exosomes is improved.

Description

Technical Field

[0001] The present invention relates to the field of exosome extraction, and in particular to an exosome low-temperature extraction device and a method for using the same. Background Art

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

[0003] Chinese patent CN218596389U discloses an exosome extraction device, including a shell and a first filter, a second filter and a tangential flow filter located in the shell and connected in sequence. A cooling element for cooling the shell is provided on the shell. A vacuum pump for generating negative pressure in the second filter and the first filter is provided on the second filter; a first peristaltic pump is provided between the second filter and the tangential flow filter; the pore sizes of the first filter, the second filter and the tangential flow filter are reduced in sequence; the exosome extraction device can not only make the entire exosome extraction process in a low temperature environment, but also effectively inhibit the activity of enzymes in the system to be extracted, so as to avoid the enzymes in the system to be extracted from hydrolyzing proteins into more impurity small molecules; there is no need to use expensive ultracentrifugation equipment, which can effectively shorten the time for extracting and separating exosomes compared with ultracentrifugation, and is more efficient.

[0004] Multiple filtration extractions of exosomes involve multiple filtration structures, pumps for transmitting liquids, and pipeline structures. The overall extraction equipment is large in size, and after the cell fluid has been filtered multiple times, the amount of liquid filtered and isolated on the filter membrane gradually decreases, and some exosomes are precipitated on the filter membrane. The exosomes cannot be completely transmitted to different filtration structures. There is a loss of exosomes during the transmission and filtration process, and some exosomes precipitated on the filter membrane also have a great impact on the filtration effect of the membrane, making it difficult to filter out some small molecules, thereby affecting the purity of the final filtered exosomes. Summary of the invention

[0005] In view of the above problems, a low-temperature extraction device for exosomes and a method for using the same are provided. The multiple filtering structures are simplified by a steering component and a liquid delivery component. When the collecting cylinder is rotated to the bottom of different filter cylinders, the cell fluid in the filter cylinder is oscillated and dispersed by an oscillation component. The problem that the secondary filtration extraction of exosomes involves multiple filtering structures, a pump body for transmitting liquid, and a pipeline structure, the volume of the overall extraction device is large, and after the cell fluid has been filtered multiple times, the amount of liquid filtered and isolated on the filter membrane gradually decreases, and some exosomes are precipitated on the filter membrane. The exosomes cannot be completely transmitted to different filtering structures. There is a loss of exosomes during the transmission and filtration process, and some exosomes precipitated on the filter membrane also have a great influence on the filtration effect of the filter membrane, making it difficult for some small molecules to be filtered out, thereby affecting the purity of the final filtered exosomes.

[0006] In order to solve the problems of the prior art, the present invention provides an exosome low-temperature extraction device, comprising an extraction frame and a bearing platform fixed on the extraction frame, the exosome extraction device also comprising a plurality of filter cartridges mounted on the bearing platform at equal angles, a collecting cartridge on the extraction frame circumferentially slidably mounted, a vacuum tube connected to the collecting cartridge, and a liquid inlet tube fixed in the filter cartridge for adding an auxiliary exosome extraction reagent; filter membranes with different pore sizes are installed in the plurality of filter cartridges; a sliding block is slidably mounted on the side wall of the collecting cartridge, a connecting ring connecting the sealing collecting cartridge and the filter cartridge is fixed on the sliding block, a bogie is fixedly connected to the collecting cartridge, a connecting lifting rod is fixedly mounted on the bogie, and a sliding block is fixed at the lifting end of the connecting lifting rod; a steering assembly for controlling the collection cartridge to rotate to the bottom of different filter cartridges is provided on the extraction frame, and a liquid conveying assembly for controlling the transmission of liquid in different filter cartridges is provided on the steering assembly.

[0007] Preferably, the steering assembly includes a driving motor fixed on the extraction frame, the output end of the driving motor is fixedly connected to a driving rotating rod, a lower gear column is fixed on the driving rotating rod, an inner gear groove rod is provided in an engaging sliding sleeve on the lower gear column, the inner gear groove rod is movably connected to the bearing platform, an upper gear column is rotatably installed on the extraction frame, the upper gear column and the inner gear groove rod are engaged and slidably connected; a shift lifting rod is fixedly installed on the steering frame, a lifting ring is fixed on the lifting end of the shift lifting rod, and the lifting ring and the inner gear groove rod are rotatably assembled.

[0008] Preferably, the liquid delivery assembly includes a support frame fixedly mounted on the inner toothed rod, a pump body is fixedly mounted on the support frame, an absorption tube for absorbing exosome liquid is connected to the pump body, and a discharge tube for discharging exosome liquid is also connected to the pump body, the absorption tube and the discharge tube respectively correspond to the positions of two groups of filter cartridges adjacent to each other in a circumference; the outlet ends of the absorption tube and the discharge tube correspond coaxially to the filter cartridge, and the length of the absorption tube is greater than the length of the discharge tube.

[0009] Preferably, the liquid inlet pipe is provided with an oscillation component for controlling the oscillation of the filter cartridge to prevent the filter membrane from being blocked by cell fluid sediment.

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

[0011] Preferably, a driving gear fixedly connected to the upper gear column is rotatably installed on the extraction frame, a plurality of driven gears meshing with the driving gear are rotatably connected on the extraction frame, 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 rail is fixedly installed on the extraction frame, a guide slide is slidably assembled on the guide rail, the guide slide and the slide column are slidably assembled, a pushing rod is rotatably connected to the guide slide, and the pushing rod is rotatably connected to a positioning ring.

[0012] Preferably, a low-temperature control component for regulating the temperature of exosome extraction is mounted on the extraction rack; the low-temperature control component comprises a heat-insulating cover mounted on the extraction rack, a cooling pipe is mounted on the heat-insulating cover, a condenser is fixed in the heat-insulating cover, a compressor is mounted on one end connected between the condenser and the cooling pipe, a throttle valve is mounted on the other end connected between the condenser and the cooling pipe, and a fan for controlling the heat dissipation of the condenser is mounted on the heat-insulating cover.

[0013] Preferably, the cooling tube is connected to a shunt cooling component for uniformly regulating the exosome extraction temperature in the filter cartridge.

[0014] Preferably, the diversion cooling assembly includes a cooling ring coaxially sleeved on the outside of the filter cylinder, a cooling branch pipe is installed in the cooling ring, a water inlet pipe and a water outlet pipe for controlling the inflow and outflow of coolant are connected through the cooling branch pipe, the water inlet pipe and the water outlet pipe are connected through the cooling pipe via a telescopic guide tube; a fixing frame is fixedly installed between the water inlet pipe, the water 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 for using an exosome low-temperature extraction device, which has the above-mentioned exosome low-temperature extraction device and includes the following steps: S1. Simplify the extraction equipment for multiple filtration. The cell fluid is introduced into a filter cartridge through a liquid inlet tube. After preliminary filtration, the valve of the filter cartridge is closed. The liquid in the filter cartridge is introduced into an adjacent filter cartridge through a liquid delivery component. The collection cartridge is controlled to move to the bottom of different filter cartridges through a steering component. Multiple filter cartridges and a group of liquid delivery components can complete multiple filtrations of the cell fluid.

[0016] S2. Oscillating the cell fluid to assist in the extraction of exosomes. When exosome particles are precipitated, the filter membrane is easily blocked. Oscillate the filter cartridge while rotating the collection cartridge to prevent the exosomes in the filter cartridge from precipitating and blocking the filter membrane.

[0017] S3. Adjust the temperature of exosome extraction. The ambient temperature of exosome extraction is precisely controlled by adjusting the temperature of the filter cartridge. The extraction temperature is maintained at 4°C to avoid degradation of exosomes during extraction. After extraction, the storage temperature of exosomes is controlled at -70°C.

[0018] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention is provided with a steering assembly and a liquid transmission assembly to simplify the equipment and can complete multiple filtering treatments of the cell fluid. After the cell fluid is filtered once, it can be transmitted to another filter cartridge through the liquid transmission assembly, and the positions of the collection cartridge and the liquid transmission assembly are switched by rotating the steering assembly, so that the cell fluid can be filtered multiple times. During the transmission of the cell fluid, the valve structure under the filter cartridge can be closed, and reagents for assisting the extraction of exosomes can be added to the filter cartridge through the liquid inlet pipe, thereby increasing the liquid volume and reducing the loss of exosomes during the continuous transmission process.

[0019] 2. The present invention is provided with an oscillation component to generate oscillation on the filter cartridge during the driving process of the steering component. The steering component drives and controls the circular rotation of the collecting cartridge. The upper tooth column in the steering component drives the driving gear to rotate, and controls the rotation of the driven gear under the gear meshing transmission action. The driven gear pushes the guide slide to move back and forth laterally through the slide column, and the guide slide controls the longitudinal movement of the positioning ring through the push rod. During the longitudinal movement, the positioning ring controls the sliding ring to move back and forth while moving up and down outside the liquid inlet pipe. The rotation of the positioning ring oscillates and strikes the filter cartridge through the oscillation rod and the oscillation hammer, so that the cell fluid in the filter cartridge is oscillated and dispersed, thereby preventing the exosomes in the cell fluid from precipitating on the filter membrane, reducing the loss during the transmission of the exosomes, and also preventing the exosomes from precipitating and clogging the filter membrane.

[0020] 3. The present invention is provided with a shunt cooling component that can evenly and effectively regulate the temperature of the filter cartridge for extracting exosomes. The coolant flowing in the cooling pipe flows into the cooling branch pipe through the telescopic guide tube and the water inlet pipe. The cooling branch pipe is sleeved on the outside of the filter cartridge to adjust the temperature around the filter cartridge at a close distance. When the positioning ring in the oscillation component moves longitudinally, the positioning ring drives the cooling ring to move up and down synchronously through the positioning frame. The up and down reciprocating movement of the cooling ring can expand the range of the cooling branch pipe to regulate the temperature of the filter cartridge, improve the effect of temperature regulation, and quickly and effectively keep the cell fluid in the filter cartridge under a low temperature environment to extract exosomes. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a schematic diagram of the three-dimensional structure of an extraction frame of an exosome low-temperature extraction device.

[0022] Figure 2 This is a schematic diagram of the three-dimensional structure of a collection tube of an exosome low-temperature extraction device.

[0023] Figure 3 This is a schematic diagram of the three-dimensional structure of a carrier platform of an exosome low-temperature extraction device.

[0024] Figure 4 This is a schematic diagram of the three-dimensional structure of a filter cartridge of an exosome low-temperature extraction device.

[0025] Figure 5 This is a schematic diagram of the three-dimensional structure of the liquid inlet tube of an exosome low-temperature extraction device.

[0026] Figure 6 This is a schematic diagram of the three-dimensional structure of the inner alveolar rod of an exosome low-temperature extraction device.

[0027] Figure 7 This is a schematic diagram of the three-dimensional structure of a vacuum tube of an exosome low-temperature extraction device.

[0028] Figure 8 This is a schematic diagram of the three-dimensional structure of the filter membrane of an exosome low-temperature extraction device.

[0029] Fig. 9 This is a schematic diagram of the three-dimensional structure of the upper tooth column of an exosome low-temperature extraction device.

[0030] Fig.10 This is a schematic diagram of the three-dimensional structure of the driving gear of an exosome low-temperature extraction device.

[0031] Fig.11 This is a schematic diagram of the three-dimensional structure of a driven gear of an exosome low-temperature extraction device.

[0032] Fig.12 This is a schematic diagram of the three-dimensional structure of a push rod of an exosome low-temperature extraction device.

[0033] Fig.13 This is a schematic diagram of the three-dimensional structure of a slip ring of an exosome low-temperature extraction device.

[0034] Fig.14 It is a schematic diagram of the three-dimensional structure of a guide slide of an exosome low-temperature extraction device.

[0035] Fig.15 This is a schematic diagram of the three-dimensional structure of a cooling tube of an exosome low-temperature extraction device.

[0036] Fig.16 This is a schematic diagram of the three-dimensional structure of a cooling ring of an exosome low-temperature extraction device.

[0037] Fig.17This is a schematic diagram of the three-dimensional structure of the cooling branch pipe of an exosome low-temperature extraction device.

[0038] The numbers in the figure are: 1. Extraction rack; 11. Carrying platform; 12. Filter cartridge; 121. Filter membrane; 13. Collection cartridge; 131. Sliding block; 132. Connecting ring; 133. Steering rack; 134. Connecting lifting rod; 14. Vacuum tube; 15. Liquid inlet pipe; 2. Steering assembly; 21. Driving motor; 22. Driving rotating rod; 23. Lower gear column; 24. Internal toothed rod; 241. Shifting lifting rod; 242. Lifting ring; 25. Upper gear column; 3. Liquid delivery assembly; 31. Support frame; 32. Pump body; 33. Absorption tube; 34. Discharge pipe; 4. Oscillation assembly; 41. S shaped slide groove; 42, slip ring; 43, positioning ring; 431, driving gear; 432, driven gear; 433, sliding column; 434, guide slide rail; 435, guide slide; 436, pushing rod; 44, oscillation rod; 45, oscillation hammer; 5, low temperature control component; 51, insulation cover; 52, cooling pipe; 53, condenser; 54, compressor; 55, throttle valve; 56, fan; 6, shunt cooling component; 61, cooling ring; 62, cooling branch pipe; 63, water inlet pipe; 64, water outlet pipe; 65, fixing frame; 651, positioning frame; 66, telescopic guide tube. DETAILED DESCRIPTION

[0039] In order to further understand the features, technical means, specific objectives and functions of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0040] See also Figure 1-Figure 9 As shown, an exosome low-temperature extraction device comprises an extraction frame 1 and a carrier platform 11 fixed on the extraction frame 1, and the exosome extraction device further comprises a plurality of filter cartridges 12 mounted on the carrier platform 11 at equal angles, a collection cartridge 13 circumferentially slidably mounted on the extraction frame 1, a vacuum tube 14 connected to the collection cartridge 13, and a liquid inlet tube 15 fixed in the filter cartridge 12 for adding an auxiliary exosome extraction reagent; filter membranes 121 with different pore sizes are installed in the plurality of filter cartridges 12; a sliding block 131 is slidably mounted on the side wall of the collection cartridge 13, and a connection seal between the collection cartridge 13 and the filter cartridge 13 is fixed on the sliding block 131. The connecting ring 132 of the filter cartridge 12 and the bogie 133 are fixedly connected to the collecting cartridge 13, and a connecting lifting rod 134 is fixedly installed on the bogie 133, and a sliding block 131 is fixed to the lifting end of the connecting lifting rod 134; the extraction frame 1 is provided with a steering assembly 2 for controlling the collection cartridge 13 to rotate to the bottom of different filter cartridges 12, and the steering assembly 2 is provided with a liquid delivery assembly 3 for controlling the transmission of liquid in different filter cartridges 12; the collecting cartridge 13 is provided with a vacuum and inflation device outside the collecting cartridge 13, which is connected to the vacuum tube 14 and is used to adjust the air pressure inside the collecting cartridge 13 so as to assist the cell fluid filtration and extraction of exosomes.

[0041] The cell fluid is input into one of the filter cartridges 12 through the liquid inlet pipe 15, and the connecting lifting rod 134 is operated to push the sliding block 131 to move upward, and the sliding block 131 drives the connecting ring 132 to move upward synchronously. The connecting ring 132 is connected and arranged between the filter cartridge 12 and the collection cartridge 13. The air pressure in the collection cartridge 13 is adjusted through the vacuum tube 14, and the valve below the filter cartridge 12 is opened. Under the filtering action of the filter membrane 121 in the filter cartridge 12, the cell fluid passes through the filter membrane 121 and enters the collection cartridge 13 below. After filtering, the valve below the filter cartridge 12 is closed, and the reagent for auxiliary exosome extraction is added to the filter cartridge 12 through the liquid inlet pipe 15 to increase the amount of solution in the filter cartridge 12. The exosomes isolated on the filter membrane 121 are better mixed in the liquid, and then the liquid transport component 3 is used to control the isolated liquid in the filter cartridge 12 to be transferred to the adjacent filter cartridge 12. The connecting lifting rod 134 is operated to control the connecting ring 132 to move down and be stored in the collecting cartridge 13. The steering component 2 is then operated to control the circular rotation of the collecting cartridge 13, so that the collecting cartridge 13 rotates to correspond to the filter cartridge 12 containing the cell fluid, and the previous cell fluid filtration steps are repeated to filter and extract the exosomes in the cell fluid again. After multiple extractions, the purity of the exosome extraction can be improved. The steering component 2 and the liquid transport component 3 can simplify the composition structure of the multiple filtration equipment, and effectively extract the exosomes in the cell fluid.

[0042] See also Figure 3-Figure 9 As shown, the steering assembly 2 includes a driving motor 21 fixed on the extraction frame 1, and the output end of the driving motor 21 is fixedly connected to a driving rotating rod 22, a lower gear column 23 is fixed on the driving rotating rod 22, and an inner tooth groove rod 24 is provided on the lower gear column 23 in a snap-fit ​​sliding sleeve, and the inner tooth groove rod 24 is movably connected to the bearing platform 11, and an upper gear column 25 is rotatably installed on the extraction frame 1, and the upper gear column 25 and the inner tooth groove rod 24 are snap-fitted and slidably connected; a shift lifting rod 241 is fixedly installed on the bogie 133, and a lifting ring 242 is fixed on the lifting end of the shift lifting rod 241, and the lifting ring 242 is rotatably assembled with the inner tooth groove rod 24.

[0043] The shift lifting rod 241 is operated to control the lifting ring 242 to drive the inner toothed rod 24 to move upward, and the inner toothed rod 24 drives the liquid delivery assembly 3 to move upward and away from the filter cylinder 12. The driving motor 21 is operated to control the driving rotating rod 22 to rotate. The driving rotating rod 22 drives the lower tooth column 23, the inner toothed rod 24 and the upper tooth column 25 to rotate synchronously. The driving rotating rod 22 drives the bogie 133 and the collecting cylinder 13 to rotate. The collecting cylinder 13 is attached to the extraction frame 1 and rotates in a circle, so that the collecting cylinder 13 rotates in a circle to the bottom of another filter cylinder 12. The inner toothed rod 24 rotates to drive the liquid delivery assembly 3 to rotate in a circle, so that the liquid delivery assembly 3 rotates to correspond to the other two groups of filter cylinders 12, so that the collecting cylinder 13, the filter cylinder 12 and the liquid delivery assembly 3 are combined into a new cell fluid filtration assembly to effectively filter and separate exosomes.

[0044] See also Figure 3-Figure 9 As shown, the liquid delivery assembly 3 includes a support frame 31 fixedly sleeved on the inner tooth groove rod 24, a pump body 32 is fixedly installed on the support frame 31, an absorption tube 33 for absorbing exosome liquid is connected to the pump body 32, and a discharge tube 34 for discharging exosome liquid is also connected to the pump body 32, the absorption tube 33 and the discharge tube 34 respectively correspond to the positions of two groups of filter cartridges 12 adjacent to each other in the circumference; the outlet ends of the absorption tube 33 and the discharge tube 34 correspond to the filter cartridge 12 coaxially, and the length of the absorption tube 33 is greater than that of the discharge tube 34; the filter membranes 121 in different filter cartridges 12 have different pore densities, and cell fluids of different molecular sizes can be filtered.

[0045] The running pump body 32 absorbs the cell fluid in the filter cartridge 12 through the absorption tube 33, and the cell fluid enters another filter cartridge 12 through the discharge tube 34, so that the cell fluid filtered and isolated in one filter cartridge 12 is transferred to the new filter cartridge 12 for re-filtration and treatment, thereby improving the purity of the extracted exosomes.

[0046] See also Figure 2-Figure 14 As shown, the liquid inlet pipe 15 is provided with an oscillation component 4 for controlling the oscillation of the filter cartridge 12 so as to prevent the filter membrane 121 from being blocked by cell fluid sediment.

[0047] When the steering component 2 controls the collection tube 13 and the liquid delivery component 3 to rotate, the oscillation component 4 can be driven to oscillate the filter tube 12 through the oscillation component 4 to disperse the cell fluid in the filter tube 12 and prevent the exosomes in the cell fluid from precipitating and clogging the filter membrane 121.

[0048] See also Figure 2-Figure 14 As shown, the oscillation assembly 4 includes an S-shaped slide groove 41 opened on the liquid inlet pipe 15, a slip ring 42 is slidably connected to the S-shaped slide groove 41, and a positioning ring 43 is rotatably assembled 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 knocking the inner wall of the filter cylinder 12 is fixedly connected to the oscillation rod 44.

[0049] The control positioning ring 43 is sleeved on the outside of the liquid inlet pipe 15 to move back and forth longitudinally, and the positioning ring 43 drives the slip ring 42 to move back and forth synchronously up and down. The slip ring 42 moves longitudinally along the direction of the S-shaped slide groove 41, and the slip ring 42 moves up and down while reciprocating. The oscillation rod 44 and the oscillation hammer 45 fixed on the slip ring 42 move longitudinally and reciprocate synchronously. The reciprocating oscillation rod 44 and the oscillation hammer 45 oscillate and strike the filter cartridge 12, so that the filter cartridge 12 vibrates, so that the particulate matter in the cell fluid in the filter cartridge 12 is dispersed, and the large molecular exosomes are prevented from being filtered and separated on the filter membrane 121.

[0050] See also Figure 4-Figure 14 As shown, a driving gear 431 fixedly connected to the upper gear column 25 is rotatably installed on the extraction frame 1, and a plurality of driven gears 432 meshing and assembled with the driving gear 431 are rotatably connected on the extraction frame 1. The number of the driven gears 432 is the same as the number of the filter cartridge 12. A sliding column 433 is eccentrically connected to the driven gear 432. A guide rail 434 is fixedly installed on the extraction frame 1, and a guide slide 435 is slidably assembled on the guide rail 434. The guide slide 435 is slidably assembled with the sliding column 433. A pushing rod 436 is rotatably connected to the guide slide 435, and the pushing rod 436 is rotatably connected to the positioning ring 43.

[0051] When the upper gear column 25 in the steering assembly 2 rotates, the upper gear column 25 drives the active gear 431 to rotate synchronously, and the driven gear 432 meshingly connected to the active gear 431 rotates accordingly, and the sliding column 433 eccentrically connected to the driven gear 432 rotates in a circle. When the sliding column 433 rotates in a circle, it pushes the guide slide 435 to reciprocate along the guide slide rail 434, and the two ends of the pushing rod 436 connected between the guide slide 435 and the positioning ring 43 rotate accordingly. The rotating pushing rod 436 pushes the positioning ring 43 which is sleeved on the outside of the liquid inlet pipe 15 to reciprocate longitudinally, thereby driving the slip ring 42 to move longitudinally and rotate along the direction of the S-shaped slide groove 41, thereby driving the oscillation rod 44 and the oscillation hammer 45 to oscillate and strike the filter cylinder 12 to disperse the cell fluid inside it.

[0052] See also Figure 1 , Figure 2 , Figure 15-17As shown, a low-temperature control component 5 for regulating the temperature of exosome extraction is mounted on the extraction rack 1; the low-temperature control component 5 includes a heat preservation cover 51 mounted on the extraction rack 1, a cooling pipe 52 is mounted on the heat preservation cover 51, a condenser 53 is fixed in the heat preservation cover 51, a compressor 54 is mounted on one end connected between the condenser 53 and the cooling pipe 52, a throttle valve 55 is mounted on the other end connected between the condenser 53 and the cooling pipe 52, and a fan 56 for controlling the heat dissipation of the condenser 53 is mounted on the heat preservation cover 51; the heat preservation 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, and the other module is slidably installed on the extraction rack 1, which can be opened for inspection of the structure in the extraction rack 1, and an inspection door is provided on the module, which can be opened to observe the structure in the extraction rack 1.

[0053] The compressor 54 controls the coolant to enter the condenser 53. The coolant in the condenser 53 is rapidly cooled down by the fan 56 during the flow process. The cooled coolant enters the cooling pipe 52 through the throttle valve 55. The cycle is repeated in this way, so that the low-temperature coolant flows in the cooling pipe 52. The low temperature in the cooling pipe 52 is transferred to the extraction rack 1, so as to keep the exosome structure extracted in the extraction rack 1 in a low-temperature state, thereby preventing the exosome from being decomposed due to the influence of temperature during the extraction of the exosomes.

[0054] See also Figure 15-17 As shown, the cooling tube 52 is connected to a shunt cooling component 6 for uniformly regulating the temperature of exosome extraction in the filter cartridge 12 ; the shunt cooling component 6 can independently affect a single filter cartridge 12 to stably maintain the temperature of the cell fluid in the filter cartridge 12 for extracting exosomes.

[0055] Part of the coolant circulating in the cooling tube 52 flows into the shunt cooling assembly 6 , and the shunt cooling assembly 6 can further adjust the ambient temperature around the filter cartridge 12 so as to maintain the temperature of the exosome extraction in the filter cartridge 12 more stably.

[0056] See also Figure 15-17 As shown, the shunt cooling assembly 6 includes a cooling ring 61 coaxially sleeved on the outside of the filter cartridge 12, a cooling branch pipe 62 is installed in the cooling ring 61, and a water inlet pipe 63 and a water outlet pipe 64 for controlling the inflow and outflow of coolant are connected to the cooling branch pipe 62, and the water inlet pipe 63 and the water outlet pipe 64 are connected to the cooling pipe 52 through a telescopic guide tube 66; a fixing frame 65 is fixedly installed between the water inlet pipe 63, the water outlet pipe 64 and the cooling ring 61, and a positioning frame 651 is fixedly assembled between the fixing frame 65 and the positioning ring 43.

[0057] The coolant circulating in the cooling pipe 52 partially enters the water inlet pipe 63 through the telescopic guide tube 66, and the coolant in the water inlet pipe 63 flows into the cooling branch pipe 62. The cooling branch pipe 62 is sleeved on the outside of the filter cartridge 12 to adjust the ambient temperature of the surrounding environment of the filter cartridge 12. The coolant flowing in the cooling branch pipe 62 finally re-enters the cooling pipe 52 through the water outlet pipe 64 and the telescopic guide tube 66 to keep the coolant circulating in the low-temperature control component 5. When the positioning ring 43 in the oscillation component 4 moves back and forth longitudinally, the positioning ring 43 drives the fixing frame 65 to move longitudinally synchronously through the positioning frame 651. The fixing frame 65 is fixed on the outside of the cooling ring 61 to control the reciprocating up and down movement of the cooling ring 61 and the cooling branch pipe 62, thereby expanding the range of the cooling branch pipe 62 to regulate the ambient temperature of the surrounding environment of the filter cartridge 12. When the fixing frame 65 moves back and forth longitudinally, the telescopic guide tube 66 automatically retracts and adjusts to keep the cooling ring 61 moving up and down to accurately regulate the temperature of the filter cartridge 12.

[0058] See also Figure 1-Figure 17 As shown, the present invention also includes a method for using an exosome low-temperature extraction device, which has the above-mentioned exosome low-temperature extraction device, comprising the following steps: S1. Simplify the extraction equipment for multiple filtration. The cell fluid is introduced into a filter cylinder 12 through the liquid inlet pipe 15. After the preliminary filtration, the valve of the filter cylinder 12 is closed. The liquid in the filter cylinder 12 is introduced into the adjacent filter cylinder 12 through the liquid delivery component 3. The collection cylinder 13 is controlled to move to the bottom of different filter cylinders 12 through the steering component 2. Multiple filter cylinders 12 and a group of liquid delivery components 3 can complete multiple filtrations of the cell fluid.

[0059] S2. Oscillating the cell fluid to assist in the extraction of exosomes. When exosome particles are precipitated, the filter membrane 121 is easily blocked. The filter cartridge 12 is oscillated during the rotation of the collection cartridge 13 to prevent the exosomes in the filter cartridge 12 from being precipitated and blocked on the filter membrane 121 .

[0060] S3. Adjust the exosome extraction temperature. The ambient temperature of the exosome extraction is precisely controlled by adjusting the temperature of the filter cartridge 12. The extraction temperature is maintained at 4°C to avoid degradation of the exosomes during extraction. After extraction, the storage temperature of the exosomes is controlled at -70°C.

[0061] The above embodiments only express one or several implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that, for those of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the attached claims.

Claims

1. A low-temperature exosome extraction device, comprising an extraction rack (1) and a carrier platform (11) fixed on the extraction rack (1), Features: The exosome extraction device further comprises a plurality of filter cartridges (12) mounted at equal angles on the carrier platform (11), a collection cartridge (13) mounted on the extraction frame (1) in a circumferentially slidable manner, a vacuum tube (14) connected to the collection cartridge (13), and a liquid inlet tube (15) fixed in the filter cartridge (12) for adding an auxiliary exosome extraction reagent; Filter membranes (121) with different pore sizes are installed in the plurality of filter cartridges (12); A sliding block (131) is slidably mounted on the side wall of the collecting cylinder (13); a connecting ring (132) connecting the sealing collecting cylinder (13) and the filtering cylinder (12) is fixed on the sliding block (131); a bogie (133) is fixedly connected to the collecting cylinder (13); a connecting lifting rod (134) is fixedly mounted on the bogie (133); and a sliding block (131) is fixed to the lifting end of the connecting lifting rod (134); The extraction rack (1) is provided with a steering assembly (2) for controlling the collection cylinder (13) to rotate to below different filter cylinders (12), and the steering assembly (2) is provided with a liquid delivery assembly (3) for controlling the transmission of liquid in different filter cylinders (12).

2. The exosome low-temperature extraction device according to claim 1, Features: The steering assembly (2) comprises a driving motor (21) fixed on the extraction frame (1), the output end of the driving motor (21) is fixedly connected to a driving rotating rod (22), a lower tooth column (23) is fixed to the driving rotating rod (22), an inner tooth groove rod (24) is provided on the lower tooth column (23) in a snap-fitting sliding sleeve, the inner tooth groove rod (24) is movably connected to the bearing platform (11), an upper tooth column (25) is rotatably mounted on the extraction frame (1), and the upper tooth column (25) is snap-fittingly slidably connected to the inner tooth groove rod (24); A shift lifting rod (241) is fixedly mounted on the bogie (133); a lifting ring (242) is fixed to the lifting end of the shift lifting rod (241); and the lifting ring (242) is rotationally assembled with the inner toothed rod (24).

3. The exosome low-temperature extraction device according to claim 2, Features: The liquid delivery assembly (3) comprises a support frame (31) fixedly sleeved on the inner toothed rod (24), a pump body (32) fixedly mounted on the support frame (31), an absorption tube (33) for absorbing exosome liquid connected to the pump body (32), and a discharge tube (34) for discharging exosome liquid connected to the pump body (32), the absorption tube (33) and the discharge tube (34) respectively corresponding to the positions of two groups of filter cartridges (12) adjacent to each other on a circumference; The outlet ends of the absorption tube (33) and the drainage tube (34) are coaxially corresponding to the filter cartridge (12), and the length of the absorption tube (33) is greater than the length of the drainage tube (34).

4. The exosome low-temperature extraction device according to claim 1, Features: The liquid inlet pipe (15) is provided with an oscillation component (4) for controlling the oscillation of the filter cartridge (12) to prevent the filter membrane (121) from being blocked by cell fluid sediment.

5. The exosome low-temperature extraction device according to claim 4, Features: The oscillating assembly (4) comprises an S-shaped slide groove (41) formed on the liquid inlet pipe (15), a slip ring (42) being slidably connected to the S-shaped slide groove (41), and a positioning ring (43) being 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 cartridge (12) is fixedly connected to the oscillating rod (44).

6. The exosome low-temperature extraction device according to claim 5, Features: A driving gear (431) fixedly connected to the upper tooth column (25) is rotatably mounted on the extraction frame (1); a plurality of driven gears (432) meshingly assembled with the driving gear (431) are rotatably connected to the extraction frame (1); the number of the driven gears (432) is the same as the number of the filter cartridge (12); a slide column (433) is eccentrically connected to the driven gear (432); a guide rail (434) is fixedly mounted on the extraction frame (1); a guide slide (435) is slidably assembled on the guide rail (434); the guide slide (435) and the slide column (433) are slidably assembled; a pushing rod (436) is rotatably connected to the guide slide (435); the pushing rod (436) is rotatably connected to the positioning ring (43).

7. The exosome low-temperature extraction device according to claim 1, Features: The extraction rack (1) is sleeved with a low-temperature control component (5) for regulating the exosome extraction temperature; The low temperature control assembly (5) comprises a heat-insulating cover (51) sleeved on the extraction rack (1), a cooling pipe (52) being mounted on the heat-insulating cover (51), a condenser (53) being fixed in the heat-insulating cover (51), a compressor (54) being mounted on one end connected between the condenser (53) and the cooling pipe (52), a throttle valve (55) being mounted on the other end connected between the condenser (53) and the cooling pipe (52), and a fan (56) for controlling heat dissipation of the condenser (53) being mounted on the heat-insulating cover (51).

8. The exosome low-temperature extraction device according to claim 7, Features: The cooling tube (52) is connected to a flow-dividing cooling component (6) for evenly regulating the exosome extraction temperature in the filter cartridge (12).

9. The exosome low-temperature extraction device according to claim 8, Features: The split flow cooling assembly (6) comprises a cooling ring (61) coaxially sleeved on the outside of the filter cartridge (12), a cooling branch pipe (62) being installed in the cooling ring (61), a water inlet pipe (63) and a water outlet pipe (64) for controlling the inflow and outflow of cooling liquid being connected through the cooling branch pipe (62), and the water inlet pipe (63) and the water outlet pipe (64) being connected through the cooling pipe (52) via a telescopic guide tube (66); A fixing frame (65) is fixedly installed between the water inlet pipe (63), the water outlet pipe (64) and the cooling ring (61), and a positioning frame (651) is fixedly assembled between the fixing frame (65) and the positioning ring (43).

10. A method for using an exosome low-temperature extraction device, It is characterized in that An exosome low-temperature extraction device applied to any one of claims 1 to 9, comprising the following steps: S1. Simplify the extraction device for multiple filtration. The cell fluid is introduced into a filter cartridge (12) through a liquid 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 a liquid delivery component (3). The collection cartridge (13) is controlled to move to the bottom of a different filter cartridge (12) through a steering component (2). Multiple filter cartridges (12) and a set of liquid delivery components (3) can complete multiple filtration of the cell fluid. S2, oscillating the cell fluid to assist in the extraction of exosomes. When exosome particles are precipitated, the filter membrane (121) is easily blocked. The filter cylinder (12) is oscillated during the rotation of the collection cylinder (13) to prevent the exosomes in the filter cylinder (12) from being precipitated and blocked on the filter membrane (121); S3. Adjust the temperature of exosome extraction. The ambient temperature of exosome extraction is precisely controlled by adjusting the temperature of the filter cartridge (12). The extraction temperature is maintained at 4°C to avoid degradation of exosomes during extraction. After extraction, the storage temperature of exosomes is controlled at -70°C.

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