Efficient preparation device for exosome production

Abstract

The application belongs to the technical field of exosome preparation, and discloses a high-efficiency preparation device for exosome production, which comprises an assembly plate, a cultivation cylinder, a filter and a peristaltic pump, the top of the assembly plate is respectively provided with a temporary storage cylinder, an enrichment and collection assembly, a tangential ultrafiltration mechanism and a reflux pipe, and the temporary storage cylinder, the enrichment and collection assembly, the tangential ultrafiltration mechanism and the reflux pipe are sequentially communicated. The elastic reset effect of the spring one is used to realize the shaking of the ultrafiltration membrane installed in the assembly ring, so that the ultrafiltration membrane is kept in a dynamic state during the process of ultrafiltration retention of exosomes, a large number of exosomes are prevented from accumulating in the local area in front of the ultrafiltration membrane, the hydraulic pressure is prevented from acting on the exosomes accumulated in the fixed area of the ultrafiltration membrane, the damage of the ultrafiltration membrane caused by compression is avoided, the accumulation of exosomes in the fixed area is reduced in cooperation with the dynamic ultrafiltration process, the hydraulic pressure and the force of exosome filtration retention are dispersed on the ultrafiltration membrane, the uniformity of stress in the ultrafiltration process is provided, and the service life is improved.

Description

Technical Field

[0001] This invention belongs to the field of exosome preparation technology, specifically a high-efficiency preparation device for exosome production. Background Technology

[0002] Exosomes are nanoscale vesicles secreted by cells that have important biological functions and are widely used in disease diagnosis, treatment and regenerative medicine. Purified exosome solutions are prepared by using enrichment and purification methods.

[0003] Existing high-efficiency exosome production apparatuses use culture tubes to cultivate exosomes and then perform coarse filtration and tangential flow ultrafiltration to concentrate and enrich the exosomes by transporting the culture mixture. However, during actual purification, the transport of hydraulic pressure and the accumulation of exosomes on one side of the ultrafiltration membrane cause damage to the membrane at the accumulation point when a certain amount of exosomes accumulates in a fixed area on one side of the ultrafiltration membrane, significantly reducing the actual service life of the ultrafiltration membrane. Furthermore, there are currently no effective means to protect the ultrafiltration membrane during use.

[0004] In addition, a large amount of liquid remains in the pipeline after the system is shut down. The liquid containing exosomes is concentrated in the pipeline and is difficult to separate. After a long period of time, it contaminates the inside of the pipeline. The open drainage method causes air pollution in the pipeline and the treatment effect is not good. Summary of the Invention

[0005] The purpose of this invention is to provide a highly efficient preparation apparatus for exosome production, so as to solve the problems mentioned in the background art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a high-efficiency preparation device for exosome production, comprising an assembly plate, a culture cylinder, a filter element, and a peristaltic delivery pump. A temporary storage cylinder, an enrichment and collection assembly, a tangential ultrafiltration mechanism, and a reflux pipe are respectively arranged above the assembly plate. The temporary storage cylinder, the enrichment and collection assembly, the tangential ultrafiltration mechanism, and the reflux pipe are sequentially connected. The lower end of the reflux pipe is connected to the culture cylinder. The peristaltic delivery pump draws mixed liquid from the culture cylinder through the filter element and introduces it into the temporary storage cylinder through a delivery pipeline assembly. The tangential ultrafiltration mechanism includes a filter cylinder body, a filter cylinder cover, and a tangential flow ultrafiltration assembly. An elastic support assembly is fixedly provided on the outer side of the tangential flow ultrafiltration assembly. The filter cylinder body and the filter cylinder cover are both fixedly sleeved on the outside of the elastic support assembly. A rotation adjustment part is rotatably sleeved within the annular gap between the filter cylinder body and the filter cylinder cover.

[0007] The elastic support assembly includes a second mounting ring, an inner ring cavity, an assembly ring, a first magnet, and a first spring. The assembly ring is rotatably sleeved in the inner ring cavity of the second mounting ring. One end of the first spring is fixed in the inner ring cavity, and the other end is fixedly connected to the assembly ring. The first magnet is fixed at equal intervals on the outer surface of the assembly ring.

[0008] The rotation adjustment unit includes a rotating ring, a second magnetic block, and a driven gear. The rotating ring rotates in the annular gap between the filter cartridge body and the filter cartridge cover. The second magnetic block is fixedly nested inside the rotating ring and, during rotation, sequentially pushes each of the first magnetic blocks on the assembly ring with magnetic force. During the rotation of the rotation adjustment unit, the magnetic force intermittently pushes the assembly ring in the elastic support assembly. A power assembly is provided above the filter cartridge body. The power assembly includes a motor, a rotating shaft, a first driving gear, and a second driving gear. The rotating shaft is fixedly connected to the output shaft of the motor. The first and second driving gears are both fixedly sleeved on the rotating shaft. The first driving gear meshes with the driven gear on the outside of the rotating ring.

[0009] Preferably, the tangential flow ultrafiltration assembly includes a connecting ring and an ultrafiltration membrane, the ultrafiltration membrane being fixedly connected in the connecting ring, the connecting ring being fixedly sleeved inside the assembly ring, and the filter cartridge being fixedly connected to the return pipe.

[0010] Preferably, the enrichment and collection assembly includes an enrichment cylinder, a mounting sleeve, a conductive curved tube, and a mounting ring. The enrichment cylinder is threaded onto the bottom of the mounting sleeve. The conductive curved tubes are symmetrically fixed to the top of the mounting sleeve and are all in communication with the mounting sleeve. The mounting ring is fitted onto the outer end of the conductive curved tube. A sealing ring is fixedly fitted onto the outer end of the conductive curved tube and is rotatably fitted into the inner wall of the mounting ring. One mounting ring is fixedly connected to the end face of the filter cartridge cover, and the other mounting ring is fixedly connected to the side of the temporary storage cylinder.

[0011] Preferably, a composite processing component is fixedly sleeved on the outer side of one of the conductive curved tubes. The power component drives the meshing composite processing component to rotate, and the composite processing component drives the enrichment and collection component to rotate. The composite processing component includes a driven gear two and a raised ring. The driven gear two is fixedly sleeved on the outer side of the conductive curved tube. The driven gear two meshes with the driving gear two. The raised ring is fixedly connected to the side of the driven gear two.

[0012] Preferably, the filter element is connected to the outlet end on the side of the cultivation cylinder, the suction end of the peristaltic pump is connected to the filter element, the delivery pipeline assembly includes an elastic guide tube and a connecting tube, the upper end of the connecting tube is fixedly connected to the temporary storage cylinder, the lower end of the connecting tube is connected to the elastic guide tube, the outlet end of the peristaltic pump is connected to the elastic guide tube, a liquid recovery section is provided below the left end of the elastic guide tube, a reciprocating pushing assembly is provided above the elastic guide tube, and the composite treatment component intermittently pushes the reciprocating pushing assembly.

[0013] Preferably, the elastic guide tube includes a tube body and an elastic connecting tube. The elastic connecting tube is fixedly connected to the upper and lower ends of the tube body. One elastic connecting tube is fixedly connected to the guide tube, and the other elastic connecting tube is fixedly connected to the outlet end of the peristaltic pump.

[0014] Preferably, the reciprocating push assembly includes a linkage, a movable rod, and a second spring. The upper end of the movable rod is fixedly connected to the linkage, and the lower end of the movable rod is movably sleeved in the adapter groove on the top of the assembly plate. One end of the second spring is fixedly connected to the lower end of the movable rod, and the other end is fixed in the adapter groove on the top of the assembly plate. One end of the linkage is located above the left end of the tube body, and the other end is located at the bottom of the protruding ring and in contact with the protruding ring.

[0015] Preferably, the liquid recovery unit includes a storage cylinder, a connecting sleeve, a sealing plug, a first tube, and a second tube. The first tube and the second tube are symmetrically fixed on the upper and lower surfaces of the connecting sleeve and are both connected to the connecting sleeve. The sealing plug is threaded into the connecting sleeve. The storage cylinder is threaded into the lower end of the second tube. The first tube is fixedly connected to the bottom of the elastic connecting tube near the left end of the tube body.

[0016] Preferably, the filter cartridge and the temporary storage cartridge are both fixed to the top of the assembly plate by bottom support rods, and the incubation cartridge and the peristaltic delivery pump are both fixedly installed on the top of the assembly plate.

[0017] The beneficial effects of this invention are as follows:

[0018] 1. This invention utilizes the combined action of a power component, a rotation adjustment unit, an elastic support component, and a tangential flow ultrafiltration component. During the ultrafiltration and retention of exosomes, the continuous rotation of the rotation adjustment unit, using magnetic repulsion, and the continuous switching of different directions during rotation, pushes the assembly ring in the elastic support component. Combined with the elastic reset effect of the spring, this causes the ultrafiltration membrane installed in the assembly ring to vibrate. This ensures that the ultrafiltration membrane is in a dynamic state during the ultrafiltration and retention of exosomes, preventing a large amount of exosomes from accumulating in a localized area on the front of the ultrafiltration membrane. It also prevents hydraulic pressure from acting on the exosomes accumulated in the fixed area of ​​the ultrafiltration membrane, thus avoiding damage to the ultrafiltration membrane due to pressure. This dynamic ultrafiltration process reduces exosome accumulation in the fixed area, ensuring that the hydraulic pressure and the force of exosome filtration and retention are distributed throughout the ultrafiltration membrane, providing uniform stress during the ultrafiltration process and improving its service life.

[0019] 2. This invention utilizes the rotation effect of the power component, in conjunction with the rotating enrichment and collection component, to ensure that when the preparation system stops supplying liquid, excess mixed liquid in the temporary storage cylinder and the enrichment and collection component continues to permeate through the ultrafiltration membrane due to density difference. The enriched liquid gradually concentrates in the enrichment cylinder. At this point, by rotating the enrichment cylinder, when the enrichment cylinder rotates to the top, the enriched liquid in the enrichment cylinder is guided out under gravity after the rotation. The enriched liquid is further introduced to one side of the ultrafiltration membrane through the conductive curved tube, realizing a secondary permeation treatment of the enriched liquid after static permeation enrichment. After the static permeation enrichment is stopped, the enriched liquid can also be subjected to secondary permeation treatment to ensure that the purity of the exosomes in the final enrichment cylinder is higher, which greatly improves the purity of the prepared enriched exosomes.

[0020] 3. This invention utilizes the power component to drive the rotation of the composite processing component, which, in conjunction with the rotation of the raised ring, achieves the reciprocating motion of the reciprocating push component. Combined with the elastic guide tube and the liquid recovery unit, after shutdown, the liquid in the pipe is collected into a storage cylinder via the liquid recovery unit. With the opening and closing of the sealing plug, the liquid is stored in the storage cylinder, and the storage cylinder is disassembled and separated under the seal at the top of the pipe, completing the liquid treatment. This effectively cleans the internal liquid while maintaining the pipe in a closed state, preventing the accumulation of exosomes and other substances in the pipe. Utilizing the elasticity of the elastic connecting tubes at both ends of the pipe, and in conjunction with the vibration treatment of the pipe by the reciprocating push component, the extrusion in the pipe is concentrated and gathered at the left end under tilted vibration, and then separated using the liquid recovery unit. This achieves the effect of thoroughly separating residual exosomes in the pipeline, while also providing pollution-free separation under a sealed pipeline condition, resulting in excellent separation performance. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of the structure of the present invention;

[0022] Figure 2This is a cross-sectional schematic diagram of the present invention;

[0023] Figure 3 This is a cross-sectional schematic diagram of the enrichment collection component and filter cartridge of the present invention;

[0024] Figure 4 This is a schematic diagram of the power assembly of the present invention;

[0025] Figure 5 This is an exploded view of the elastic support component and the tangential flow ultrafiltration component of the present invention;

[0026] Figure 6 This is an exploded view of the rotating adjustment part, filter cartridge body, and filter cartridge cover of the present invention;

[0027] Figure 7 This is a schematic diagram of the composite processing component of the present invention;

[0028] Figure 8 This is an exploded view of the enrichment and collection component of the present invention;

[0029] Figure 9 This is a schematic diagram of the reciprocating drive component of the present invention;

[0030] Figure 10 This is a schematic diagram showing the connection between the elastic guide tube and the liquid collection and recovery section of the present invention;

[0031] Figure 11 This is a cross-sectional schematic diagram of the liquid recovery section of the present invention.

[0032] In the diagram: 1. Assembly plate; 2. Culture cylinder; 3. Filter element; 4. Peristaltic transfer pump; 5. Flexible guide tube; 51. Tube body; 52. Flexible connecting tube; 6. Conducting tube; 7. Temporary storage cylinder; 8. Filter cylinder body; 9. Return tube; 10. Filter cylinder cover; 11. Enrichment and collection assembly; 111. Enrichment cylinder; 112. Mounting sleeve; 113. Conducting curved tube; 114. Mounting ring one; 12. Tangential flow ultrafiltration assembly; 121. Connecting ring; 122. Ultrafiltration membrane; 13. Flexible support assembly; 131. Mounting ring two; 132. Inner ring cavity; 133. Assembly ring; 134. Magnetic block 135. Spring 1; 14. Power assembly; 141. Motor; 142. Rotating shaft; 143. Drive gear 1; 144. Drive gear 2; 15. Composite processing component; 151. Driven gear 2; 152. Protruding ring; 16. Rotation adjustment part; 161. Rotating ring; 162. Magnetic block 2; 163. Driven gear 1; 17. Reciprocating push assembly; 171. Linkage component; 172. Moving rod; 173. Spring 2; 18. Liquid recovery part; 181. Storage cylinder; 182. Connecting sleeve; 183. Sealing plug; 184. Pipe No. 1; 185. Pipe No. 2. Detailed Implementation

[0033] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0034] like Figures 1 to 11 As shown, this embodiment of the invention provides a high-efficiency preparation device for exosome production, including an assembly plate 1, a culture tube 2, and a filter element 3 (the filter element 3 is a conventional structure, specifically a TFF ultrafiltration membrane with a pore size greater than or equal to 0). The assembly plate 1 is equipped with a 0.18μm filter (filtering cells to the outside while allowing exosomes to pass through) and a peristaltic transport pump 4. Above the assembly plate 1 are a temporary storage cylinder 7, an enrichment and collection assembly 11, a tangential ultrafiltration mechanism, and a reflux pipe 9. The temporary storage cylinder 7, enrichment and collection assembly 11, tangential ultrafiltration mechanism, and reflux pipe 9 are sequentially connected. The lower end of the reflux pipe 9 is connected to the culture cylinder 2. The peristaltic transport pump 4 draws the mixed liquid from the culture cylinder 2 through the filter element 3 and introduces it into the temporary storage cylinder 7 through the transport pipeline assembly. The tangential ultrafiltration mechanism includes a filter cylinder body 8, a filter cylinder cover 10, and a tangential flow ultrafiltration assembly 12. An elastic support assembly 13 is fixedly provided on the outer side of the tangential flow ultrafiltration assembly 12. The filter cylinder body 8 and the filter cylinder cover 10 are both fixedly sleeved on the outside of the elastic support assembly 13. A rotation adjustment part 16 is rotatably sleeved within the annular gap between the filter cylinder body 8 and the filter cylinder cover 10. 13 includes a mounting ring 131, an inner annular cavity 132, an assembly ring 133, a magnetic block 134, and a spring 135. The assembly ring 133 is rotatably sleeved in the inner annular cavity 132 of the mounting ring 131. One end of the spring 135 is fixed in the inner annular cavity 132, and the other end is fixedly connected to the assembly ring 133. The magnetic blocks 134 are fixed at equal intervals on the outer surface of the assembly ring 133. The rotation adjustment part 16 includes a rotating ring 161, a magnetic block 162, and a driven gear 163. The rotating ring 161 rotates in the annular gap between the filter cartridge body 8 and the filter cartridge cover 10. The magnetic block 162 is fixedly nested inside the rotating ring 161 and, during rotation, sequentially pushes each magnetic block 134 on the assembly ring 133 with magnetic force. During the rotation of the rotation adjustment part 16, the magnetic force intermittently pushes the assembly ring 133 in the elastic support assembly 13.

[0035] The assembly ring 133 is provided with sealing gaskets (not shown in the figure) on both sides in the inner ring cavity 132 to ensure that the contact surface between the side of the assembly ring 133 and the inner wall of the inner ring cavity 132 has a dynamic seal, and to ensure that the assembly ring 133 maintains a dynamic sealing effect when sliding, so as to avoid seepage and leakage along the contact surface between the assembly ring 133 and the inner ring cavity 132.

[0036] The filter cartridge 8 is equipped with a power assembly 14 above it. The power assembly 14 includes a motor 141, a rotating shaft 142, a first drive gear 143, and a second drive gear 144. The rotating shaft 142 is fixedly connected to the output shaft of the motor 141. The first drive gear 143 and the second drive gear 144 are both fixedly sleeved on the rotating shaft 142. The first drive gear 143 meshes with the driven gear 163 on the outside of the rotating ring 161. The tangential flow ultrafiltration assembly 12 includes a connecting ring 121 and an ultrafiltration membrane 122. The ultrafiltration membrane 122 (the ultrafiltration membrane 122 is a TFF ultrafiltration membrane, and it is an existing structure. The pore size of the TFF ultrafiltration membrane is less than or equal to 0.03μm to prevent exosomes from passing through and to ensure that exosomes are retained after two ultrafiltrations) is fixedly connected in the connecting ring 121. The connecting ring 121 is fixedly sleeved inside the assembly ring 133. The filter cartridge 8 is fixedly connected to the return pipe 9.

[0037] By utilizing the power assembly 14 to provide rotational power, the inner annular cavity 132 is opened inside the mounting ring 131, and an annular space is reserved for the movement of the outer side of the assembly ring 133, ensuring that the assembly ring 133 can be offset in different directions in the inner annular cavity 132.

[0038] Example 1: During preparation, as exosomes are produced in culture tube 2 and mixed in, peristaltic pump 4 is started, the mixed liquid in culture tube 2 is drawn out, and cells are filtered out by filter element 3. The filtered exosome mixture is input into temporary storage tube 7 through delivery pipeline assembly. The mixed liquid in temporary storage tube 7 is guided to enrichment and collection assembly 11 and introduced into the interior of filter tube cover 10 and filter tube body 8. The exosomes are intercepted by ultrafiltration membrane 122. The filtered liquid flows back to culture tube 2 along return pipe 9. During ultrafiltration, power assembly 14 is started. Drive gear 143 rotates and drives meshing driven gear 163 to rotate, driving rotating ring 161 to rotate along the annular gap between filter tube cover 10 and filter tube body 8, and driving magnetic block 162 to rotate. When magnetic block 162 rotates to the magnetic block inside mounting ring 131, After the corresponding position of 134 is reached, the magnetic blocks 162 and 134 are positioned with their magnetic poles facing each other. Under the action of repulsive force, the magnetic blocks 162 push the magnetic blocks 134 to move. The assembly ring 133 where the magnetic blocks 134 are located moves in the inner ring cavity 132 and pulls the corresponding spring 135. As the magnetic blocks 162 rotates around, they continuously push each magnetic block 134 to move, causing the assembly ring 133 to continuously switch directions in space. Combined with the elasticity of the spring 135, the assembly ring 133 causes the internally installed ultrafiltration membrane 122 to vibrate continuously, causing the exosomes attached to the ultrafiltration membrane 122 to detach. In the actual preparation process, vibration is started and performed intermittently as needed. During long-term preparation, vibration is started and performed for a period of time to complete the disturbance during filtration and avoid the negative effects caused by continuous long-term vibration treatment.

[0039] First, by utilizing the combined action of the power assembly 14, the rotation adjustment unit 16, the elastic support assembly 13, and the tangential flow ultrafiltration assembly 12, during the ultrafiltration retention of exosomes, the rotation adjustment unit 16 continuously rotates, utilizing magnetic repulsion, and continuously switches different directions to push the assembly ring 133 in the elastic support assembly 13. Combined with the elastic reset effect of the spring 135, this causes the ultrafiltration membrane 122 installed in the assembly ring 133 to vibrate. This ensures that the ultrafiltration membrane 122 is in a dynamic state during the ultrafiltration retention of exosomes, preventing a large amount of exosomes from accumulating in a localized area on the front of the ultrafiltration membrane 122, avoiding hydraulic pressure acting on the exosomes accumulated in the fixed area of ​​the ultrafiltration membrane 122, and preventing damage to the ultrafiltration membrane 122 under pressure. This dynamic ultrafiltration process reduces the accumulation of exosomes in the fixed area, ensuring that the hydraulic pressure and the force of exosome filtration retention are distributed throughout the ultrafiltration membrane 122, providing uniform force during the ultrafiltration process and improving its service life.

[0040] The enrichment and collection assembly 11 includes an enrichment cylinder 111, a mounting sleeve 112, a connecting curved tube 113, and a mounting ring 114. The enrichment cylinder 111 is threaded onto the bottom of the mounting sleeve 112. The connecting curved tubes 113 are symmetrically fixed to the top of the mounting sleeve 112 and are all connected to the mounting sleeve 112. The mounting ring 114 is sleeved on the outer end of the connecting curved tube 113. A sealing ring is fixedly sleeved on the outer end of the connecting curved tube 113. The sealing ring is rotatably sleeved in the inner wall of the mounting ring 114. One mounting ring 114 is fixedly connected to the end face of the filter cartridge cover 10. An installation ring 114 is fixedly connected to the side of the temporary storage cylinder 7. A composite processing component 15 is fixedly sleeved on the outer side of a conductive curved tube 113. The power component 14 drives the meshing composite processing component 15 to rotate. The composite processing component 15 drives the enrichment and collection component 11 to rotate. The composite processing component 15 includes a driven gear 151 and a raised ring 152. The driven gear 151 is fixedly sleeved on the outside of the conductive curved tube 113. The driven gear 151 meshes with the driving gear 144. The raised ring 152 is fixedly connected to the side of the driven gear 151.

[0041] The enrichment and collection component 11 achieves the enrichment and collection of exosomes. After the system is shut down, it works with the density difference on both sides of the ultrafiltration membrane 122 to achieve static permeation of excess liquid on one side, ensuring that the enriched liquid is finally concentrated in the enrichment cylinder 111. After removing the enrichment cylinder 111, the prepared and purified exosomes are obtained. The connecting tube 113 achieves rotation under dynamic sealing in the mounting rings 114 on both sides through the sealing rings. Through the control of the gear ratio, the actual enrichment cylinder 111 maintains slow rotation to achieve buffering under slow rotation. Specifically, depending on the requirements, 4-5 rotations are performed during the secondary permeation after dynamic ultrafiltration. At this time, the secondary permeation effect is optimal and the impact on the enriched exosomes is small.

[0042] Example 2: After the initial ultrafiltration enrichment is completed, as the peristaltic pump 4 stops working, the liquid in the system stops flowing and transporting. The power component 14 is kept running, the drive gear 144 rotates synchronously, and drives the meshing composite processing component 15 to rotate. The meshing driven gear 151 rotates, and drives the guide tube 113 to rotate around the inside of the mounting ring 114, and drives the enrichment cylinder 111 to rotate. When the enrichment cylinder 111 rotates to the top, the mixed liquid stored in the enrichment cylinder 111 is input into the filter cover 10 again through the guide tube 113 under the action of gravity. Under the gravity guidance after the flip, the mixed liquid flows and acts on the front of the ultrafiltration membrane 122, completing the further filtration and interception of the mixed liquid. As the enrichment cylinder 111 continues to rotate, the mixed liquid in the enrichment collection component 11 impacts the ultrafiltration membrane 122 intermittently under the guidance of gravity, and completes the supplementary interception and filtration treatment after shutdown.

[0043] First, by utilizing the rotation effect of the power component 14 again, in conjunction with the rotating enrichment and collection component 11, when the preparation system stops supplying liquid, as the excess mixed liquid in the temporary storage cylinder 7 and the enrichment and collection component 11 continues to permeate through the ultrafiltration membrane 122 under the density difference, the enriched liquid gradually concentrates in the enrichment cylinder 111. At this time, by rotating the enrichment cylinder 111, when the enrichment cylinder 111 rotates to the top, the enriched liquid in the enrichment cylinder 111 is guided out under the gravity after the inversion, and further introduced into one side of the ultrafiltration membrane 122 through the connecting curved tube 113, realizing the secondary permeation treatment of the enriched liquid after static permeation enrichment. After the static permeation enrichment is stopped, the enriched liquid can also be subjected to secondary permeation treatment, ensuring that the purity of the exosomes in the final enrichment cylinder 111 is higher, which greatly improves the purity of the prepared enriched exosomes.

[0044] The filter element 3 is connected to the outlet end on the side of the cultivation cylinder 2, the suction end of the peristaltic pump 4 is connected to the filter element 3, the conveying pipeline assembly includes an elastic guide tube 5 and a guide tube 6, the upper end of the guide tube 6 is fixedly connected to the temporary storage cylinder 7, the lower end of the guide tube 6 is connected to the elastic guide tube 5, the outlet end of the peristaltic pump 4 is connected to the elastic guide tube 5, a liquid recovery section 18 is provided below the left end of the elastic guide tube 5, and a reciprocating pushing assembly 17 is provided above the elastic guide tube 5. The composite processing component 15 intermittently pushes the reciprocating pushing assembly 17.

[0045] Filter element 3 is an existing component with an internal filter membrane to achieve coarse filtration, isolate cells, and ensure the output of exosomes. The elastic guide tube 5 enables the output of the liquid after coarse filtration.

[0046] The elastic guide pipe 5 includes a pipe body 51 and an elastic connecting pipe 52. The elastic connecting pipe 52 is fixedly connected to the upper and lower ends of the pipe body 51. One elastic connecting pipe 52 is fixedly connected to the guide pipe 6, and the other elastic connecting pipe 52 is fixedly connected to the outlet end of the peristaltic pump 4.

[0047] The elastic connecting tube 52 allows for elastic deformation, ensuring that the tube body 51 vibrates back and forth at the top, and under the action of the force above the multiple ends, it achieves asymmetrical elastic deformation, ensuring that the left end of the tube body 51 is lower when vibrating, and guiding the accumulated liquid in the tube body 51 to flow to the left end.

[0048] The reciprocating push assembly 17 includes a linkage 171, a movable rod 172, and a second spring 173. The upper end of the movable rod 172 is fixedly connected to the linkage 171, and the lower end of the movable rod 172 is movably sleeved in the adapter groove on the top of the assembly plate 1. One end of the second spring 173 is fixedly connected to the lower end of the movable rod 172, and the other end is fixed in the adapter groove on the top of the assembly plate 1. One end of the linkage 171 is located above the left end of the tube body 51, and the other end is located at the bottom of the protruding ring 152 and contacts the protruding ring 152.

[0049] The reciprocating push assembly 17 is pushed by the protruding ring 152 to achieve reciprocating up and down movement, and the elastic cooperation of the second spring 173 achieves elastic reset.

[0050] The liquid recovery unit 18 includes a storage cylinder 181, a connecting sleeve 182, a sealing plug 183, a first pipe 184, and a second pipe 185. The first pipe 184 and the second pipe 185 are symmetrically fixed on the upper and lower surfaces of the connecting sleeve 182 and are both connected to the connecting sleeve 182. The sealing plug 183 is threaded into the connecting sleeve 182. The storage cylinder 181 is threaded into the lower end of the second pipe 185. The first pipe 184 is fixedly connected to the bottom of the elastic connecting pipe 52 near the left end of the pipe body 51.

[0051] The liquid recovery unit 18 achieves the sealing and opening of the left end elastic connecting tube 52 through the action of the middle sealing plug 183, and works with the bottom storage tube 181 to ensure that the tube body 51 is also isolated from the ambient air when it is opened, and to ensure that the tube body 51 is sealed when the storage tube 181 is removed, so as to avoid air pollution when cleaning the liquid.

[0052] The filter cartridge 8 and the temporary storage cartridge 7 are both fixed to the top of the assembly plate 1 by bottom support rods, and the incubation cartridge 2 and the peristaltic transfer pump 4 are both fixedly installed on the top of the assembly plate 1.

[0053] Example 3: After the exosome enrichment, purification, and separation are completed, the power component 14 is kept running, driving the protruding ring 152 in the composite processing component 15 to rotate. During the rotation, the reciprocating pushing component 17 is intermittently squeezed. The top end of the linkage component 171 moves downward when squeezed by the protrusion, driving the movable rod 172 to compress the second spring 173, and causing the lower end of the linkage component 171 to move downward and squeeze the left end of the tube body 51. With the reciprocating pushing of the protruding ring 152, the linkage component 171 moves up and down back and forth, causing the tube body 51 to vibrate back and forth. When the left end of the tube body 51 is pressed down, the two The elastic connecting tubes 52 at both ends are elastically deformed, and the push near the left end of the tube body 51 causes the left end of the tube body 51 to have a greater elastic deformation, making the tube body 51 appear as a left-low and right-high state. With the reciprocating vibration, the residual liquid in the tube body 51 is gradually guided to converge into the elastic connecting tube 52 at the left end. The sealing plug 183 in the liquid recovery section 18 is opened, and the liquid is introduced into the storage cylinder 181 along the first tube 184 and the second tube 185. The sealing plug 183 is rotated back to its original position, and the storage cylinder 181 is rotated out from the bottom of the second tube 185 to achieve liquid separation.

[0054] First, by reusing the power assembly 14 to drive the rotation of the composite processing component 15, in conjunction with the rotation of the protruding ring 152, the reciprocating motion of the reciprocating push assembly 17 is achieved. Combined with the elastic guide tube 5 and the liquid recovery unit 18, after shutdown, the liquid in the pipe body 51 is collected into the storage cylinder 181 by the liquid recovery unit 18. With the opening and closing of the sealing plug 183, the liquid is stored in the storage cylinder 181. Then, with the top of the pipe body 51 sealed, the storage cylinder 181 is disassembled and separated, completing the liquid treatment and preserving the liquid. The system effectively cleans the internal liquid while the tube body 51 is closed, preventing the accumulation of exosomes and other substances in the tube body 51. Utilizing the elasticity of the elastic connecting tubes 52 at both ends of the tube body 51, and in conjunction with the reciprocating push assembly 17 to vibrate the tube body 51, the squeezed liquid in the tube body 51 is concentrated and gathered at the left end under the tilted vibration of the left end of the tube body 51. Separation is achieved using the liquid recovery section 18, which has the effect of completely separating residual exosomes in the pipeline. At the same time, it also has the effect of pollution-free separation under the pipeline's sealed state, resulting in a good separation effect.

[0055] The working principle and usage process of this invention are as follows: During preparation, as exosomes are produced in culture tube 2 and mixed in culture tube 2, the peristaltic delivery pump 4 is started, the mixed liquid in culture tube 2 is drawn out, and the cells are filtered out by filter element 3. The filtered exosome mixture is input into temporary storage tube 7 through delivery pipeline assembly. The mixed liquid in temporary storage tube 7 is guided to enrichment and collection assembly 11 and introduced into the interior of filter tube cover 10 and filter tube body 8. The exosomes are intercepted by ultrafiltration membrane 122. The filtered liquid flows back to culture tube 2 along return pipe 9. During ultrafiltration, power assembly 14 is started. The driving gear 143 rotates and drives the meshing driven gear 163 to rotate, driving the rotating ring 161 along the ring between filter tube cover 10 and filter tube body 8. The gap rotates, causing the second magnetic block 162 to rotate. When the second magnetic block 162 rotates to the position corresponding to the first magnetic block 134 inside the second mounting ring 131, the repulsive force caused by the opposing magnetic poles of the second magnetic block 162 and the first magnetic block 134 pushes the first magnetic block 134 to move. The assembly ring 133 where the first magnetic block 134 is located moves in the inner ring cavity 132 and pulls the corresponding spring 135. As the second magnetic block 162 rotates around, it continuously pushes each first magnetic block 134 to move, causing the assembly ring 133 to continuously switch the direction of displacement in space. Combined with the elasticity of the spring 135, the assembly ring 133 causes the internally installed ultrafiltration membrane 122 to vibrate continuously, and the exosomes attached to the ultrafiltration membrane 122 detach.

[0056] After the initial ultrafiltration enrichment is completed, the peristaltic pump 4 stops working, the liquid in the system stops flowing and the power component 14 is kept running. The second drive gear 144 rotates synchronously and drives the meshing composite treatment component 15 to rotate. The second driven gear 151 rotates and drives the connecting tube 113 to rotate around the inside of the mounting ring 114, and drives the enrichment cylinder 111 to rotate. When the enrichment cylinder 111 rotates to the top, the mixed liquid stored in the enrichment cylinder 111 is input into the filter cover 10 again through the connecting tube 113 under the action of gravity. Under the gravity guidance after the flip, the mixed liquid flows and acts on the front of the ultrafiltration membrane 122 to complete the further filtration and interception of the mixed liquid. As the enrichment cylinder 111 continues to rotate, the mixed liquid in the enrichment collection component 11 intermittently impacts the ultrafiltration membrane 122 under the guidance of gravity, and completes the supplementary interception and filtration treatment after shutdown.

[0057] After the exosome enrichment, purification, and separation are completed, the power assembly 14 is kept running, driving the raised ring 152 in the composite processing component 15 to rotate. During the rotation, the component 17 is intermittently squeezed and reciprocated. When the raised part squeezes, the top end of the linkage 171 moves downward, driving the movable rod 172 to compress the spring 173. This causes the lower end of the linkage 171 to move downward and squeeze the left end of the tube 51. With the reciprocating push of the raised ring 152, the linkage 171 moves up and down back and forth, causing the tube 51 to vibrate back and forth. When the left end of the tube 51 is pressed down, the elastic ends... All connecting tubes 52 are elastically deformed, and the push near the left end of the tube body 51 causes the left end of the tube body 51 to have a greater elastic deformation, making the tube body 51 appear as a left-low and right-high state. With the reciprocating vibration, the residual liquid in the tube body 51 is gradually guided to converge into the elastic connecting tube 52 at the left end. The sealing plug 183 in the liquid recovery section 18 is opened, and the liquid is introduced into the storage cylinder 181 along the first tube 184 and the second tube 185. The sealing plug 183 is rotated back to its original position, and the storage cylinder 181 is rotated out from the bottom of the second tube 185 to achieve liquid separation.

[0058] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. An efficient preparation device for exosome production, comprising an assembly plate (1), a cultivation cylinder (2), a filter (3) and a peristaltic pump (4), the upper portion of the assembly plate (1) is respectively provided with a temporary storage cylinder (7), an enrichment collection assembly (11), a tangential ultrafiltration mechanism and a reflux pipe (9), the temporary storage cylinder (7), the enrichment collection assembly (11), the tangential ultrafiltration mechanism and the reflux pipe (9) are sequentially communicated, the lower end of the reflux pipe (9) is communicated with the cultivation cylinder (2), the peristaltic pump (4) sucks mixed liquid in the cultivation cylinder (2) through the filter (3) and introduces the mixed liquid into the temporary storage cylinder (7) through a delivery pipeline assembly, characterized in that: The tangential ultrafiltration mechanism comprises a filter cartridge body (8), a filter cartridge cover (10) and a tangential flow ultrafiltration assembly (12), an elastic support assembly (13) is fixedly arranged on the outer side of the tangential flow ultrafiltration assembly (12), the filter cartridge body (8) and the filter cartridge cover (10) are fixedly sleeved on the outer side of the elastic support assembly (13), a rotating adjusting part (16) is rotatably sleeved in the annular gap between the filter cartridge body (8) and the filter cartridge cover (10), The elastic support assembly (13) comprises a mounting ring two (131), an inner ring cavity (132), an assembly ring (133), a magnetic block one (134) and a spring one (135), the assembly ring (133) is rotatably sleeved in the inner ring cavity (132) of the mounting ring two (131), one end of the spring one (135) is fixed in the inner ring cavity (132), and the other end is fixedly connected with the assembly ring (133), the magnetic block one (134) is fixedly arranged on the outer side of the assembly ring (133) at equal intervals, The rotating adjusting part (16) comprises a rotating ring (161), a magnetic block two (162) and a driven gear one (163), the rotating ring (161) rotates in the annular gap between the filter cartridge body (8) and the filter cartridge cover (10), the magnetic block two (162) is fixedly nested in the rotating ring (161), and sequentially pushes each magnetic block one (134) on the assembly ring (133) by magnetic force in the rotating process, the rotating adjusting part (16) intermittently pushes the assembly ring (133) in the elastic support assembly (13) by magnetic force in the rotating process, a power assembly (14) is arranged above the filter cartridge body (8), the power assembly (14) comprises a motor (141), a rotating shaft (142), a driving gear one (143) and a driving gear two (144), the rotating shaft (142) is fixedly connected with the output shaft of the motor (141), the driving gear one (143) and the driving gear two (144) are fixedly sleeved on the rotating shaft (142), and the driving gear one (143) is in meshing connection with the driven gear one (163) on the outer side of the rotating ring (161).

2. The efficient preparation device for exosome production according to claim 1, characterized in that: The tangential flow ultrafiltration assembly (12) comprises a connecting ring (121) and an ultrafiltration membrane (122), the ultrafiltration membrane (122) is fixedly connected in the connecting ring (121), the connecting ring (121) is fixedly sleeved in the assembly ring (133), and the filter cartridge body (8) is fixedly communicated with the return pipe (9).

3. The efficient preparation device for exosome production according to claim 1, characterized in that: The enrichment collecting assembly (11) comprises an enrichment cylinder (111), a mounting sleeve (112), a lead-through elbow (113) and a mounting ring one (114), the enrichment cylinder (111) is threadedly mounted at the bottom of the mounting sleeve (112), the lead-through elbows (113) are symmetrically fixed at the top of the mounting sleeve (112) and communicate with the mounting sleeve (112), the mounting ring one (114) is sleeved at the outer end of the lead-through elbow (113), the outer end of the lead-through elbow (113) is fixedly sleeved with a sealing snap ring, the sealing snap ring is rotatably sleeved in the inner wall of the mounting ring one (114), one mounting ring one (114) is fixedly connected to the end surface of the filter cylinder cover (10), and the other mounting ring one (114) is fixedly connected to the side surface of the temporary storage cylinder (7).

4. The efficient preparation device for exosome production according to claim 3, characterized in that: The outer side surface of one lead-through elbow (113) is fixedly sleeved with a composite processing piece (15), the power assembly (14) drives the engaged composite processing piece (15) to rotate, the composite processing piece (15) drives the enrichment collecting assembly (11) to rotate, the composite processing piece (15) comprises a driven gear two (151) and a protruding ring (152), the driven gear two (151) is fixedly sleeved at the outer side of the lead-through elbow (113), the driven gear two (151) is in meshing connection with the driving gear two (144), and the protruding ring (152) is fixedly connected to the side surface of the driven gear two (151).

5. The efficient preparation device for exosome production according to claim 4, characterized in that: The filter piece (3) communicates with the outlet end of the side surface of the cultivation cylinder (2), the suction end of the peristaltic pump (4) communicates with the filter piece (3), the conveying pipeline assembly comprises an elastic flow guide pipe piece (5) and a lead-through pipe (6), the upper end of the lead-through pipe (6) is fixedly connected with the temporary storage cylinder (7), the lower end of the lead-through pipe (6) communicates with the elastic flow guide pipe piece (5), the outlet end of the peristaltic pump (4) communicates with the elastic flow guide pipe piece (5), and the lower portion of the left end of the elastic flow guide pipe piece (5) is provided with a liquid accumulation recovery portion (18). The upper portion of the elastic flow guide pipe piece (5) is provided with a reciprocating pushing assembly (17), and the composite processing piece (15) intermittently pushes the reciprocating pushing assembly (17).

6. The efficient preparation device for exosome production according to claim 5, characterized in that: The elastic flow guide pipe piece (5) comprises a pipe body (51) and elastic connecting pipes (52), the elastic connecting pipes (52) are fixedly connected to the upper and lower ends of the pipe body (51), one elastic connecting pipe (52) is fixedly connected with the lead-through pipe (6), and the other elastic connecting pipe (52) is fixedly connected with the outlet end of the peristaltic pump (4).

7. The efficient preparation device for exosome production according to claim 6, characterized in that: The reciprocating pushing assembly (17) comprises a linkage (171), a movable rod (172) and a spring (173), the upper end of the movable rod (172) is fixedly connected with the linkage (171), the lower end of the movable rod (172) is movably sleeved in the adaptive groove on the top of the assembling plate (1), one end of the spring (173) is fixedly connected with the lower end of the movable rod (172), and the other end is fixed in the adaptive groove on the top of the assembling plate (1), one end of the linkage (171) is located above the left end of the pipe body (51), and the other end is located at the bottom of the protruding ring (152) and is in contact with the protruding ring (152).

8. The efficient preparation device for exosome production according to claim 7, characterized in that: The effusion recovery part (18) comprises a storage cylinder (181), a communication sleeve (182), a sealing plug (183), a No. 1 pipe (184) and a No. 2 pipe (185), the No. 1 pipe (184) and the No. 2 pipe (185) are symmetrically fixed on the upper and lower surfaces of the communication sleeve (182) and are in communication with the communication sleeve (182), the sealing plug (183) is threadedly sleeved in the communication sleeve (182), the storage cylinder (181) is threadedly sleeved at the lower end of the No. 2 pipe (185), and the No. 1 pipe (184) is fixedly communicated at the bottom of the elastic connecting pipe (52) close to the left end of the pipe body (51).

9. The efficient preparation device for exosome production according to claim 1, characterized in that: The filter cylinder body (8) and the temporary storage cylinder (7) are both fixed on the top of the assembling plate (1) through bottom support rods, and the cultivation cylinder (2) and the peristaltic pump (4) are both fixedly installed on the top of the assembling plate (1).

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