Efficient preparation device for exosome production

Abstract

The invention belongs to the technical field of exosome preparation, and discloses an efficient preparation device for exosome production, the efficient preparation device comprises an assembly plate, a cultivation cylinder, a filter part and a peristaltic conveying pump, a temporary storage cylinder, an enrichment collection assembly, a tangential ultrafiltration mechanism and a return pipe are arranged above the assembly plate; and the temporary storage cylinder, the enriching and collecting assembly, the tangential ultrafiltration mechanism and the return pipe are communicated in sequence. Through the elastic reset effect of the first spring, shaking of the ultrafiltration membrane installed in the assembly ring is achieved, it is guaranteed that the ultrafiltration membrane is in a dynamic state in the exosome ultrafiltration interception process, it is avoided that a large number of exosomes are accumulated in the local area of the front face of the ultrafiltration membrane, hydraulic pressure is prevented from acting on the exosomes accumulated in the ultrafiltration membrane fixing area, and the effect of the hydraulic pressure on the exosomes is avoided. According to the present invention, with the dynamic ultrafiltration process, the exosome accumulation in the fixed area is reduced, the hydraulic force and the exosome filtration interception acting force are dispersed at all positions of the ultrafiltration membrane, the stress uniformity in the ultrafiltration process is provided, and the service life is prolonged.

Description

Technical Field

[0001] The present invention belongs to the technical field of exosome preparation, and specifically relates to a high-efficiency preparation device for exosome production. Background Art

[0002] Exosomes are nanoscale vesicles secreted by cells, which have important biological functions and are widely used in fields such as disease diagnosis, treatment, and regenerative medicine. Through enrichment and purification means, a purified exosome solution can be prepared.

[0003] In the existing high-efficiency preparation device for exosome production, during use, exosomes are cultivated in a cultivation cylinder, and the cultivated mixed liquid is transported for rough cell filtration and tangential flow ultrafiltration interception treatment to achieve the centralized enrichment of exosomes. However, during the actual purification and preparation process, the conveying hydraulic pressure and the enriched exosomes accumulate on one side of the ultrafiltration membrane. When a certain amount of exosomes accumulates in a fixed area on one side of the ultrafiltration membrane, combined with the hydraulic pressure, the ultrafiltration membrane at the stacking and pressing position is damaged, greatly reducing the actual service life of the ultrafiltration membrane, and there is currently no effective means to protect the ultrafiltration membrane during use.

[0004] In addition, a large amount of liquid remains in the pipeline of the preparation system after shutdown. The liquid containing exosomes accumulates in the pipeline and is difficult to separate, and it contaminates the inside of the pipeline after long-term retention. The open liquid discharge method causes air pollution in the pipeline and the treatment effect is not good. Summary of the Invention

[0005] The purpose of the present invention is to provide a high-efficiency preparation device for exosome production to solve the problems raised in the above background art.

[0006] To achieve the above purpose, the present invention provides the following technical solution: A high-efficiency preparation device for exosome production, including an assembly plate, a cultivation cylinder, a filtering component, and a peristaltic pump. Above the assembly plate, there are respectively a temporary storage cylinder, an enrichment and collection component, a tangential ultrafiltration mechanism, and a return pipe. The temporary storage cylinder, the enrichment and collection component, the tangential ultrafiltration mechanism, and the return pipe are connected in sequence. The lower end of the return pipe is connected to the cultivation cylinder. The peristaltic pump sucks the mixed liquid in the cultivation cylinder through the filtering component and introduces it into the temporary storage cylinder through a conveying pipeline component. The tangential ultrafiltration mechanism includes a filter cylinder body, a filter cylinder cover, and a tangential flow ultrafiltration component. An elastic support component is fixedly arranged on the outer side of the tangential flow ultrafiltration component. The filter cylinder body and the filter cylinder cover are both fixedly sleeved on the outer side of the elastic support component. A rotation adjustment part is rotatably sleeved in the annular gap between the filter cylinder body and the filter cylinder cover.

[0007] The elastic support assembly includes a mounting ring, an inner ring cavity, a fitting ring, a first magnet block, and a first spring. The fitting ring is rotatably sleeved in the inner ring cavity of the mounting ring. One end of the first spring is fixed in the inner ring cavity, and the other end is fixedly connected to the fitting ring. The first magnet blocks are evenly fixed on the outer side surface of the fitting ring.

[0008] The rotation adjustment part includes a rotating ring, a second magnet block, and a first driven gear. The rotating ring rotates in the annular gap between the filter cylinder body and the filter cylinder cover. The second magnet block is fixedly nested inside the rotating ring and magnetically pushes each first magnet block on the fitting ring in sequence during rotation. The fitting ring in the elastic support assembly is intermittently magnetically pushed during the rotation of the rotation adjustment part.

[0009] Preferably, a power assembly is provided above the filter cylinder 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 driving gear and the second driving gear are both fixedly sleeved on the rotating shaft. The first driving gear is meshed and connected to the first driven gear on the outer side of the rotating ring.

[0010] Preferably, the tangential flow ultrafiltration assembly includes a connecting ring and an ultrafiltration membrane. The ultrafiltration membrane is fixedly connected in the connecting ring. The connecting ring is fixedly sleeved inside the fitting ring. The filter cylinder body is fixedly communicated with the reflux pipe.

[0011] Preferably, the enrichment and collection assembly includes an enrichment cylinder, a mounting sleeve, a conduction curved pipe, and a mounting ring. The enrichment cylinder is threadedly mounted at the bottom of the mounting sleeve. The conduction curved pipes are symmetrically fixed on the top of the mounting sleeve and are both communicated with the mounting sleeve. The mounting ring is sleeved at the outer end of the conduction curved pipe. A sealing clamping ring is fixedly sleeved at the outer end of the conduction curved pipe, and the sealing clamping ring is rotatably sleeved in the inner wall of the mounting ring. One of the mounting rings is fixedly communicated with the end face of the filter cylinder cover, and the other mounting ring is fixedly communicated with the side surface of the temporary storage cylinder.

[0012] Preferably, a composite treatment part is fixedly sleeved on the outer side surface of one of the conduction curved pipes. The power assembly drives the engaged composite treatment part to rotate. The composite treatment part drives the enrichment and collection assembly to rotate. The composite treatment part includes a second driven gear and a protruding ring. The second driven gear is fixedly sleeved on the outer side of the conduction curved pipe. The second driven gear is meshed and connected to the second driving gear. The protruding ring is fixedly connected to the side surface of the second driven gear.

[0013] Preferably, the filter element is communicated with the outlet end on the side of the cultivation cylinder, the suction end of the peristaltic pump is communicated with the filter element, the conveying pipeline assembly includes an elastic diversion pipe fitting and a conduction pipe, the upper end of the conduction pipe is fixedly communicated with the temporary storage cylinder, the lower end of the conduction pipe is communicated with the elastic diversion pipe fitting, the outlet end of the peristaltic pump is communicated with the elastic diversion pipe fitting, a liquid accumulation recovery part is arranged below the left end of the elastic diversion pipe fitting, a reciprocating pushing assembly is arranged above the elastic diversion pipe fitting, and the composite treatment part intermittently pushes the reciprocating pushing assembly.

[0014] Preferably, the elastic diversion pipe fitting includes a pipe body and an elastic connecting pipe, the elastic connecting pipes are fixedly connected to the upper and lower ends of the pipe body, one elastic connecting pipe is fixedly connected to the conduction pipe, and the other elastic connecting pipe is fixedly connected to the outlet end of the peristaltic pump.

[0015] Preferably, the reciprocating pushing assembly includes a linkage part, a movable rod and a second spring. The upper end of the movable rod is fixedly connected to the linkage part. The lower end of the movable rod is movably sleeved in the fitting groove at 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 fitting groove at the top of the assembly plate. One end of the linkage part is located above the left end of the pipe body, and the other end is located at the bottom of the convex ring and contacts the convex ring.

[0016] Preferably, the liquid accumulation recovery part includes a storage cylinder, a connecting sleeve, a sealing plug, a first pipe and a second pipe. The first pipe and the second pipe are symmetrically fixed on the upper and lower surfaces of the connecting sleeve and are both communicated with the connecting sleeve. The sealing plug is threadedly sleeved in the connecting sleeve. The storage cylinder is threadedly sleeved at the lower end of the second pipe. The first pipe is fixedly communicated with the bottom of the elastic connecting pipe near the left end of the pipe body.

[0017] Preferably, both the filter cylinder body and the temporary storage cylinder are fixed to the top of the assembly plate through bottom support rods, and the cultivation cylinder and the peristaltic pump are both fixedly installed on the top of the assembly plate.

[0018] The beneficial effects of the present invention are as follows:

[0019] 1. By utilizing the combined action of the power component, the rotational adjustment part, the elastic support component, and the tangential flow ultrafiltration component, when ultrafiltration and retention of exosomes are carried out, through the continuous rotation of the rotational adjustment part, the magnetic repulsion effect is utilized, and during the rotation process, the assembly ring in the elastic support component is continuously pushed in different directions, and in cooperation with the elastic reset effect of the first spring, the shaking of the ultrafiltration membrane installed in the assembly ring is realized, ensuring that the ultrafiltration membrane is in a dynamic state during the ultrafiltration and retention of exosomes, avoiding the accumulation of a large number of exosomes in a local area on the front side of the ultrafiltration membrane, avoiding the hydraulic pressure acting on the exosomes accumulated in the fixed area of the ultrafiltration membrane, and avoiding damage to the ultrafiltration membrane caused by compression. In cooperation with the dynamic ultrafiltration process, the accumulation of exosomes in the fixed area is reduced, ensuring that the hydraulic pressure and the acting force of exosome filtration and retention are dispersed throughout the ultrafiltration membrane, providing the force uniformity during the ultrafiltration process, and improving the service life.

[0020] 2. By once again utilizing the effect of the rotation of the power component and cooperating with the rotationally installed enrichment and collection component, when the preparation system stops transporting liquid, as the excess mixed liquid in the temporary storage cylinder and the enrichment and collection component continues to permeate through the ultrafiltration membrane under the density difference, the enriched liquid gradually concentrates in the enrichment cylinder. At this time, by realizing the rotation of the enrichment cylinder, when the enrichment cylinder rotates to the upper side, the enriched liquid in the enrichment cylinder is drained out under the gravity after flipping, and further, the enriched liquid is introduced to one side of the ultrafiltration membrane through the conduction curved pipe, realizing the secondary permeation treatment of the enriched liquid after static permeation enrichment. After static permeation enrichment during shutdown, the enriched liquid can also be subjected to secondary permeation treatment, ensuring that the purity of the exosomes in the finally obtained enrichment cylinder is higher, and greatly improving the purity of the prepared and enriched exosomes.

[0021] 3. By once again utilizing the rotation of the power component to drive the rotation of the composite treatment part and cooperating with the rotation of the convex ring, the reciprocating action of the reciprocating push component is realized, and in cooperation with the elastic diversion pipe fitting and the liquid accumulation recovery part, after shutdown, the liquid accumulation in the pipe body is collected into the storage cylinder through the liquid accumulation recovery part, and in cooperation with the opening and closing of the sealing plug, the liquid accumulation is first stored in the storage cylinder, and the storage cylinder is disassembled and separated under the sealing of the top of the pipe body to complete the treatment of the liquid accumulation, effectively cleaning the internal liquid accumulation while maintaining the closed state of the pipe body, avoiding the accumulation of substances such as exosomes in the pipe body. Utilizing the elastic action of the elastic connecting pipes at both ends of the pipe body and cooperating with the vibration treatment of the pipe body by the reciprocating push component, when the left end of the pipe body vibrates obliquely, the extrusion in the pipe body is concentrated and converged at the left end, and separation is realized by the liquid accumulation recovery part, having the effect of thoroughly separating the residual exosome liquid accumulation in the pipeline, and at the same time having pollution-free separation under the sealed state of the pipeline, with good separation effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 is a schematic structural diagram of the present invention;

[0023] Figure 2Cross-sectional schematic diagram of the present invention;

[0024] Figure 3 Cross-sectional schematic diagram of the enrichment and collection component and the filter cylinder body of the present invention;

[0025] Figure 4 Schematic diagram of the power component of the present invention;

[0026] Figure 5 Explosion schematic diagram of the elastic support component and the tangential flow ultrafiltration component of the present invention;

[0027] Figure 6 Explosion schematic diagram of the rotation adjustment part, the filter cylinder body and the filter cylinder cover of the present invention;

[0028] Figure 7 Schematic diagram of the composite treatment part of the present invention;

[0029] Figure 8 Explosion schematic diagram of the enrichment and collection component of the present invention;

[0030] Figure 9 Schematic diagram of the reciprocating push component of the present invention;

[0031] Figure 10 Connection schematic diagram of the elastic diversion pipe fitting and the liquid accumulation recovery part of the present invention;

[0032] Figure 11 Cross-sectional schematic diagram of the liquid accumulation recovery part of the present invention.

[0033] In the figure: 1. Assembly plate; 2. Cultivation cylinder; 3. Filter element; 4. Peristaltic pump; 5. Elastic diversion pipe fitting; 51. Pipe body; 52. Elastic connecting pipe; 6. Conducting pipe; 7. Temporary storage cylinder; 8. Filter cylinder body; 9. Return pipe; 10. Filter cylinder cover; 11. Enrichment and collection component; 111. Enrichment cylinder; 112. Installation sleeve; 113. Conducting curved pipe; 114. Installation ring; 12. Tangential flow ultrafiltration component; 121. Connecting ring; 122. Ultrafiltration membrane; 13. Elastic support component; 131. Installation ring; 132. Inner cavity; 133. Assembly ring; 134. Magnet 1; 135. Spring 1; 14. Power component; 141. Motor; 142. Rotating shaft; 143. Driving gear 1; 144. Driving gear 2; 15. Composite treatment part; 151. Driven gear 2; 152. Protruding ring; 16. Rotation adjustment part; 161. Rotating ring; 162. Magnet 2; 163. Driven gear 1; 17. Reciprocating push component; 171. Linkage part; 172. Moving rod; 173. Spring 2; 18. Liquid accumulation recovery part; 181. Storage cylinder; 182. Connecting sleeve; 183. Sealing plug; 184. No. 1 pipe; 185. No. 2 pipe. Detailed implementation manners

[0034] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0035] As Figures 1 to 11 shown, an efficient preparation device for exosome production provided by an embodiment of the present invention includes an assembly plate 1, a cultivation cylinder 2, a filter member 3 (the filter member 3 is a conventional structure, specifically a TFF ultrafiltration membrane with a pore size greater than or equal to 0.18 μm, which filters cells to the outside and allows exosomes to pass through), and a peristaltic pump 4. Above the assembly plate 1, there are respectively provided a temporary storage cylinder 7, an enrichment and collection assembly 11, a tangential ultrafiltration mechanism, and a return pipe 9. The temporary storage cylinder 7, the enrichment and collection assembly 11, the tangential ultrafiltration mechanism, and the return pipe 9 are connected in sequence. The lower end of the return pipe 9 is connected to the cultivation cylinder 2. The peristaltic pump 4 sucks the mixed liquid in the cultivation cylinder 2 through the filter member 3 and introduces it into the temporary storage cylinder 7 through a 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. Both the filter cylinder body 8 and the filter cylinder cover 10 are fixedly sleeved on the outside of the elastic support assembly 13. A rotation adjustment part 16 is rotatably sleeved in the annular gap between the filter cylinder body 8 and the filter cylinder cover 10. The elastic support assembly 13 includes a mounting ring 131, an inner ring cavity 132, an assembly ring 133, a first magnetic block 134, and a first spring 135. The assembly ring 133 is rotatably sleeved in the inner ring cavity 132 of the mounting ring 131. One end of the first spring 135 is fixed in the inner ring cavity 132, and the other end is fixedly connected to the assembly ring 133. The first magnetic blocks 134 are equidistantly fixed on the outer side of the assembly ring 133. The rotation adjustment part 16 includes a rotation ring 161, a second magnetic block 162, and a first driven gear 163. The rotation ring 161 rotates in the annular gap between the filter cylinder body 8 and the filter cylinder cover 10. The second magnetic block 162 is fixedly nested inside the rotation ring 161 and sequentially magnetically pushes each of the first magnetic blocks 134 on the assembly ring 133 during rotation. During the rotation of the rotation adjustment part 16, the assembly ring 133 in the elastic support assembly 13 is intermittently magnetically pushed.

[0036] Sealing gaskets (not shown in the figure) are provided on both sides of the assembly ring 133 in the inner ring cavity 132 to ensure dynamic sealing of the contact surface between the side surface of the assembly ring 133 and the inner wall of the inner ring cavity 132, and to ensure that the dynamic sealing effect is maintained when the assembly ring 133 slides, so as to avoid leakage along the contact surface between the assembly ring 133 and the inner ring cavity 132.

[0037] Among them, a power assembly 14 is provided above the filter cylinder body 8. The power assembly 14 includes a motor 141, a rotating shaft 142, a first driving gear 143 and a second driving gear 144. The rotating shaft 142 is fixedly connected to the output shaft of the motor 141. Both the first driving gear 143 and the second driving gear 144 are fixedly sleeved on the rotating shaft 142. The first driving gear 143 is meshed and connected with a first driven gear 163 on the outer side 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 is an existing structure, and the pore diameter of the TFF ultrafiltration membrane is less than or equal to 0.03 μm to prevent exosomes from passing through and ensure the retention of exosomes 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 cylinder body 8 is fixedly communicated with the reflux pipe 9.

[0038] By using the power assembly 14 to provide rotational power, an inner ring cavity 132 is opened inside the mounting ring 131, and an annular space for the movement of the outer side of the assembly ring 133 is reserved to ensure that the assembly ring 133 can offset in different directions in the inner ring cavity 132.

[0039] Example 1: During preparation, as exosomes are produced in the cultivation cylinder 2 and mixed in the cultivation cylinder 2, start the peristaltic pump 4, suck out the mixed liquid in the cultivation cylinder 2, filter out the cells through the filter element 3, and input the filtered exosome mixture into the temporary storage cylinder 7 through the conveying pipeline assembly. The mixed liquid entering the temporary storage cylinder 7 is guided into the enrichment and collection assembly 11 and then introduced into the inside of the filter cylinder cover 10 and the filter cylinder body 8. The exosomes are intercepted by the ultrafiltration membrane 122, and the filtered liquid flows back to the cultivation cylinder 2 along the reflux pipe 9. During ultrafiltration, start the power assembly 14. The first driving gear 143 rotates and drives the meshed first driven gear 163 to rotate, driving the rotating ring 161 to rotate along the annular gap between the filter cylinder cover 10 and the filter cylinder body 8, and driving the second magnetic block 162 to rotate. When the second magnetic block 162 rotates to the corresponding position of the first magnetic block 134 inside the mounting ring 131, with the same magnetic pole of the second magnetic block 162 and the first magnetic block 134 facing each other, the repulsive force causes the second magnetic block 162 to push 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 first spring 135. As the second magnetic block 162 rotates around, it continuously magnetically pushes each first magnetic block 134 to move, causing the assembly ring 133 to offset in different directions in space. With the elasticity of the first spring 135, the assembly ring 133 drives the internally installed ultrafiltration membrane 122 to vibrate continuously, and the exosomes attached to the ultrafiltration membrane 122 are detached. During the actual preparation process, start and perform vibration treatment intermittently according to requirements. During long-term preparation, start and perform vibration for a period of time to complete the disturbance during filtration and avoid the negative impacts caused by continuous long-term vibration treatment.

[0040] First, by utilizing the cooperative action of the power component 14, the rotation adjustment part 16, the elastic support component 13, and the tangential flow ultrafiltration component 12, when performing ultrafiltration interception of exosomes, through the continuous rotation of the rotation adjustment part 16, using the magnetic repulsion force, and continuously switching the pushing direction of the assembly ring 133 in the elastic support component 13 during the rotation process, and cooperating with the elastic reset effect of the first spring 135, the shaking of the ultrafiltration membrane 122 installed in the assembly ring 133 is realized, ensuring that the ultrafiltration membrane 122 is in a dynamic state during the ultrafiltration interception of exosomes, avoiding the accumulation of a large number of exosomes in the local area on the front side of the ultrafiltration membrane 122, avoiding the hydraulic pressure acting on the exosomes accumulated in the fixed area of the ultrafiltration membrane 122, avoiding damage to the ultrafiltration membrane 122 caused by compression, cooperating with the dynamic ultrafiltration process, reducing the accumulation of exosomes in the fixed area, ensuring that the liquid pressure and the acting force of exosome filtration and interception are dispersed everywhere on the ultrafiltration membrane 122, providing the force uniformity during the ultrafiltration process, and improving the service life.

[0041] Among them, the enrichment and collection component 11 includes an enrichment cylinder 111, a mounting sleeve 112, a conduction curved pipe 113, and a mounting ring 114. The enrichment cylinder 111 is threadedly installed at the bottom of the mounting sleeve 112. The conduction curved pipes 113 are symmetrically fixed at the top of the mounting sleeve 112 and are both communicated with the mounting sleeve 112. The mounting ring 114 is sleeved on the outer ends of the conduction curved pipes 113. A sealing clamping ring is fixedly sleeved on the outer ends of the conduction curved pipes 113, and the sealing clamping ring is rotatably sleeved in the inner wall of the mounting ring 114. One mounting ring 114 is fixedly communicated with the end face of the filter cylinder cover 10, and the other mounting ring 114 is fixedly communicated with the side surface of the temporary storage cylinder 7. A composite treatment part 15 is fixedly sleeved on the outer side surface of one conduction curved pipe 113. The power component 14 drives the engaged composite treatment part 15 to rotate, and the composite treatment part 15 drives the enrichment and collection component 11 to rotate. The composite treatment part 15 includes a driven gear two 151 and a convex ring 152. The driven gear two 151 is fixedly sleeved on the outer side of the conduction curved pipe 113, the driven gear two 151 is meshed and connected with the driving gear two 144, and the convex ring 152 is fixedly connected to the side surface of the driven gear two 151.

[0042] The enrichment and collection component 11 realizes the enrichment and collection of exosomes, and after the system stops, it cooperates with the density difference on both sides of the ultrafiltration membrane 122 to realize the static penetration of the excess liquid on one side, ensuring that the enrichment liquid is finally concentrated in the enrichment cylinder 111, and obtaining the prepared and purified exosomes after removing the enrichment cylinder 111. The conduction curved pipe 113 realizes rotation under dynamic sealing in the two mounting rings 114 through the sealing clamping ring, and through the control of the gear ratio, the actual enrichment cylinder 111 maintains slow rotation to realize buffering under slow flipping. Specifically, according to requirements, 4 - 5 rotation treatments are performed during the secondary penetration after the dynamic ultrafiltration is completed. At this time, the best secondary penetration effect is obtained, and the impact on the enriched exosomes is small.

[0043] Embodiment 2: After the preliminary ultrafiltration enrichment is completed, as the peristaltic pump 4 stops working, the liquid in the system stops flowing and transporting. Keep the power assembly 14 started, the second driving gear 144 rotates synchronously, and drives the engaged composite processing part 15 to rotate. The engaged second driven gear 151 rotates, and drives the conducting curved pipe 113 to rotate inside the mounting ring 114, and drives the enrichment cylinder 111 to rotate. When the enrichment cylinder 111 rotates to the upper side, under the action of gravity of the mixed liquid stored in the enrichment cylinder 111, it is input into the filter cartridge cover 10 again through the conducting curved pipe 113 on one side. Under the action of gravity diversion after flipping, the mixed liquid flows and acts on the front side of the ultrafiltration membrane 122, completing the further filtration and interception of the mixed liquid. As the enrichment cylinder 111 rotates continuously, the mixed liquid in the enrichment collection assembly 11 impacts the ultrafiltration membrane 122 intermittently under the guidance of gravity, and completes the supplementary interception and filtration treatment after shutdown.

[0044] First, by reusing the rotation effect of the power assembly 14 and cooperating with the rotationally mounted enrichment collection assembly 11, when the liquid transportation in the preparation system stops, as the excess mixed liquid in the temporary storage cylinder 7 and the enrichment collection assembly 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 realizing the rotation of the enrichment cylinder 111, when the enrichment cylinder 111 rotates to the upper side, the enriched liquid in the enrichment cylinder 111 is drained out under the gravity after flipping, and further the enriched liquid is introduced to one side of the ultrafiltration membrane 122 through the conducting curved pipe 113, realizing the secondary permeation treatment of the enriched liquid after static permeation enrichment. After the static permeation enrichment during shutdown, the secondary permeation treatment of the enriched liquid can still be carried out, ensuring that the purity of the exosomes in the final obtained enrichment cylinder 111 is higher, and greatly improving the purity of the prepared and enriched exosomes.

[0045] Among them, the filter element 3 is communicated with the outlet end on the side of the cultivation cylinder 2. The suction end of the peristaltic pump 4 is communicated with the filter element 3. The transportation pipeline assembly includes an elastic diversion pipe fitting 5 and a conducting pipe 6. The upper end of the conducting pipe 6 is fixedly communicated with the temporary storage cylinder 7. The lower end of the conducting pipe 6 is communicated with the elastic diversion pipe fitting 5. The outlet end of the peristaltic pump 4 is communicated with the elastic diversion pipe fitting 5. A liquid accumulation recovery part 18 is arranged below the left end of the elastic diversion pipe fitting 5. A reciprocating pushing assembly 17 is arranged above the elastic diversion pipe fitting 5. The composite processing part 15 intermittently pushes the reciprocating pushing assembly 17.

[0046] The filter element 3 is an existing component, with a filter membrane inside, realizing rough filtration, isolating cells, ensuring the output of exosomes, and the elastic diversion pipe fitting 5 realizes the output of the liquid after rough filtration.

[0047] Among them, the elastic diversion pipe fitting 5 includes 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 to the conduction pipe 6, and the other elastic connecting pipe 52 is fixedly connected to the outlet end of the peristaltic pump 4.

[0048] The elastic connecting pipe 52 allows its elastic deformation to ensure that the pipe body 51 vibrates reciprocally up and down at the top. Under the action force above the multi-end, it realizes asymmetric elastic deformation to ensure that the left end of the pipe body 51 is lower when vibrating, guiding the accumulated liquid in the pipe body 51 to converge to the left end.

[0049] Among them, the reciprocating pushing assembly 17 includes a linkage member 171, a movable rod 172, and a second spring 173. The upper end of the movable rod 172 is fixedly connected to the linkage member 171. The lower end of the movable rod 172 is movably sleeved in the fitting groove at 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 fitting groove at the top of the assembly plate 1. One end of the linkage member 171 is located above the left end of the pipe body 51, and the other end is located at the bottom of the raised ring 152 and contacts the raised ring 152.

[0050] The reciprocating pushing assembly 17 bears the push of the raised ring 152 to realize reciprocating up and down movement, and the elastic cooperation of the second spring 173 realizes elastic reset.

[0051] Among them, the accumulated liquid recovery part 18 includes a storage cylinder 181, a communication 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 communication sleeve 182 and are both communicated 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 second pipe 185. The first pipe 184 is fixedly communicated with the bottom of the elastic connecting pipe 52 near the left end of the pipe body 51.

[0052] The accumulated liquid recovery part 18 realizes the sealing and opening of the left-end elastic connecting pipe 52 through the action of the middle sealing plug 183. Cooperating with the bottom storage cylinder 181, it ensures that the pipe body 51 is also isolated from the ambient air when opened, and ensures the sealing of the pipe body 51 when the storage cylinder 181 is taken out, avoiding air pollution during the cleaning of the accumulated liquid.

[0053] Among them, both the filter cylinder body 8 and the temporary storage cylinder 7 are fixed to the top of the assembly plate 1 through bottom support rods. The cultivation cylinder 2 and the peristaltic pump 4 are both fixedly installed on the top of the assembly plate 1.

[0054] Embodiment 3: After the exosome enrichment, purification, and separation are completed, the power component 14 is kept running to drive the convex ring 152 in the composite processing component 15 to rotate. During the rotation process, the reciprocating pushing component 17 is intermittently squeezed. When the convex part squeezes the top end of the linkage 171, it moves downward, driving the movable rod 172 to compress the second spring 173, and causing the lower end of the linkage 171 to move downward and squeeze the left end of the push tube 51. With the reciprocating push of the convex ring 152, the linkage 171 moves up and down reciprocally, causing the tube 51 to vibrate reciprocally. When the left end of the tube 51 is pressed and moves downward, both elastic connecting tubes 52 at both ends elastically deform. And under the push near the left end of the tube 51, the elastic deformation amount of the left end of the tube 51 is larger, making the overall state of the tube 51 lower on the left and higher on the right. With the cooperation of the reciprocating vibration, the residual liquid in the tube 51 is gradually guided and converged into the elastic connecting tube 52 at the left end. The operator opens the sealing plug 183 in the liquid recovery part 18, and the liquid is introduced into the storage cylinder 181 along the first tube 184 and the second tube 185. Then, the sealing plug 183 is rotated and reset, and the storage cylinder 181 is rotated and taken out from the bottom of the second tube 185 to achieve liquid separation.

[0055] First, by using the power component 14 to drive the rotation of the composite processing component 15 again, with the rotation of the convex ring 152, the reciprocating movement of the reciprocating pushing component 17 is realized. And with the cooperation of the elastic diversion pipe fitting 5 and the liquid recovery part 18, after the machine stops, the liquid in the tube 51 is collected into the storage cylinder 181 through the liquid recovery part 18. With the opening and closing of the sealing plug 183, the liquid is stored in the storage cylinder 181 first, and the storage cylinder 181 is disassembled and separated under the sealing of the top of the tube 51 to complete the treatment of the liquid. The effective cleaning of the internal liquid in the tube 51 is completed while keeping the tube 51 in a closed state, avoiding the accumulation of substances such as exosomes in the tube 51. By using the elastic effect of the elastic connecting tubes 52 at both ends of the tube 51 and cooperating with the vibration treatment of the tube 51 by the reciprocating pushing component 17, when the left end of the tube 51 vibrates obliquely, the liquid squeezed in the tube 51 is concentrated and converged at the left end, and the separation is achieved by using the liquid recovery part 18. It has the effect of completely separating the residual exosome liquid in the pipeline, and at the same time, it also has pollution-free separation under the sealed state of the pipeline, and the separation effect is good.

[0056] Working principle and usage process of the present invention: During preparation, as exosomes are produced in the culture cylinder 2 and mixed in the culture cylinder 2, the peristaltic pump 4 is started, and the mixed liquid in the culture cylinder 2 is sucked out and filtered by the filter element 3 to remove cells. The filtered exosome mixture is input into the temporary storage cylinder 7 through the conveying pipeline assembly. The mixed liquid entering the temporary storage cylinder 7 is guided to the enrichment and collection assembly 11 and introduced into the interior of the filter cylinder cover 10 and the filter cylinder body 8. The exosomes are intercepted by the ultrafiltration membrane 122, and the filtered liquid flows back to the culture cylinder 2 along the return pipe 9. During ultrafiltration, the power assembly 14 is started, the first driving gear 143 rotates and drives the engaged first driven gear 163 to rotate, driving the rotating ring 161 to rotate along the annular gap between the filter cylinder cover 10 and the filter cylinder body 8, and driving the second magnetic block 162 to rotate. When the second magnetic block 162 rotates to the corresponding position of the first magnetic block 134 inside the mounting ring 131, with the same-pole setting of the second magnetic block 162 and the first magnetic block 134, the repulsive force causes the second magnetic block 162 to push the first magnetic block 134 to move. The mounting ring 133 where the first magnetic block 134 is located moves in the inner ring cavity 132 and pulls the corresponding first spring 135. As the second magnetic block 162 rotates around, each first magnetic block 134 is continuously magnetically pushed to move, causing the mounting ring 133 to offset in different directions in space. With the elasticity of the first spring 135, the mounting ring 133 drives the internally installed ultrafiltration membrane 122 to vibrate continuously, and the exosomes attached to the ultrafiltration membrane 122 are detached;

[0057] After the preliminary ultrafiltration and enrichment are completed, as the peristaltic pump 4 stops working, the liquid in the system stops flowing and transporting. The power assembly 14 remains started, the second driving gear 144 rotates synchronously, driving the engaged composite processing part 15 to rotate, the engaged second driven gear 151 rotates, driving the conduction curved pipe 113 to rotate around the inside of the mounting ring 114, and driving the enrichment cylinder 111 to rotate. When the enrichment cylinder 111 rotates to the upper position, under the action of gravity of the mixed liquid stored in the enrichment cylinder 111, it is input into the filter cylinder cover 10 again through the conduction curved pipe 113 on one side. Under the action of gravity diversion after flipping, the mixed liquid flows and acts on the front surface of the ultrafiltration membrane 122, completing the further filtration and interception of the mixed liquid. As the enrichment cylinder 111 rotates continuously, the mixed liquid in the enrichment and collection assembly 11 impacts the ultrafiltration membrane 122 intermittently under the guidance of gravity, and the supplementary interception and filtration treatment after shutdown are completed;

[0058] After the enrichment, purification and separation of exosomes are completed, keep the power component 14 activated to drive the rotation of the convex ring 152 in the composite processing component 15. During the rotation process, intermittently squeeze and reciprocally push the reciprocating component 17. The top end of the linkage 171 moves downward when squeezed by the convex part, drives the movable rod 172 to compress the second spring 173, and makes the lower end of the linkage 171 move downward to squeeze the left end of the push tube body 51. With the reciprocating push of the convex ring 152, the linkage 171 moves up and down reciprocally, and the tube body 51 vibrates reciprocally. When the left end of the tube body 51 is pressed and moves downward, both elastic connecting tubes 52 at both ends elastically deform, and the elastic deformation amount of the left end of the tube body 51 is larger under the push near the left end of the tube body 51, making the overall state of the tube body 51 lower on the left and higher on the right. With the cooperation of the reciprocating vibration effect, the residual liquid in the tube body 51 is gradually guided and gathered into the elastic connecting tube 52 at the left end. Operate to open the sealing plug 183 in the liquid collection and recovery part 18, and the liquid flows along the first tube 184 and the second tube 185 into the storage cylinder 181. Rotate and reset the sealing plug 183, and rotate and remove the storage cylinder 181 from the bottom of the second tube 185 to achieve liquid separation.

[0059] Although the embodiments of the present invention have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principle and spirit of the present invention. The scope of the present invention 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 element (3) and a peristaltic pump (4). Above the assembly plate (1), there are respectively a temporary storage cylinder (7), an enrichment and collection assembly (11), a tangential ultrafiltration mechanism and a return pipe (9). The temporary storage cylinder (7), the enrichment and collection assembly (11), the tangential ultrafiltration mechanism and the return pipe (9) are connected in sequence. The lower end of the return pipe (9) is connected to the cultivation cylinder (2). The peristaltic pump (4) sucks the mixed liquid in the cultivation cylinder (2) through the filter element (3) and imports it into the temporary storage cylinder (7) through the conveying pipeline assembly. It is characterized in that: The tangential ultrafiltration mechanism includes a filter cylinder body (8), a filter cylinder cover (10), and a tangential flow ultrafiltration component (12). An elastic support component (13) is fixedly arranged on the outer side surface of the tangential flow ultrafiltration component (12). The filter cylinder body (8) and the filter cylinder cover (10) are both fixedly sleeved on the outer side of the elastic support component (13). A rotating adjustment part (16) is rotatably sleeved in the annular gap between the filter cylinder body (8) and the filter cylinder cover (10). The elastic support component (13) includes a mounting ring (131), an inner ring cavity (132), an assembly ring (133), a first magnetic block (134), and a first spring (135). The assembly ring (133) is rotatably sleeved in the inner ring cavity (132) of the mounting ring (131). One end of the first spring (135) is fixed in the inner ring cavity (132), and the other end is fixedly connected to the assembly ring (133). The first magnetic blocks (134) are fixedly arranged at equal intervals on the outer side surface of the assembly ring (133). The rotating adjustment part (16) includes a rotating ring (161), a second magnetic block (162), and a first driven gear (163). The rotating ring (161) rotates in the annular gap between the filter cylinder body (8) and the filter cylinder cover (10). The second magnetic block (162) is fixedly nested inside the rotating ring (161), and sequentially magnetically pushes each of the first magnetic blocks (134) on the assembly ring (133) during rotation. During the rotation of the rotating adjustment part (16), the assembly ring (133) in the elastic support component (13) is intermittently magnetically pushed.

2. The high-efficiency preparation device for exosome production according to claim 1, wherein: A power component (14) is arranged above the filter cylinder body (8). The power component (14) includes a motor (141), a rotating shaft (142), a first driving gear (143), and a second driving gear (144). The rotating shaft (142) is fixedly connected to the output shaft of the motor (141). The first driving gear (143) and the second driving gear (144) are both fixedly sleeved on the rotating shaft (142). The first driving gear (143) is meshed and connected with the first driven gear (163) on the outer side of the rotating ring (161).

3. The highly efficient preparation device for exosome production according to claim 2, wherein: The tangential flow ultrafiltration component (12) includes 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 inside the assembly ring (133). The filter cylinder body (8) is fixedly communicated with a reflux pipe (9).

4. The high-efficiency preparation device for exosome production according to claim 1, wherein: The enrichment and collection component (11) includes an enrichment cylinder (111), a mounting sleeve (112), a conducting curved pipe (113) and a mounting ring (114). The enrichment cylinder (111) is threadedly mounted at the bottom of the mounting sleeve (112). The conducting curved pipes (113) are symmetrically fixed at the top of the mounting sleeve (112) and are both communicated with the mounting sleeve (112). The mounting ring (114) is sleeved at the outer ends of the conducting curved pipes (113). A sealing clamping ring is fixedly sleeved at the outer ends of the conducting curved pipes (113), and the sealing clamping ring is rotatably sleeved in the inner wall of the mounting ring (114). One of the mounting rings (114) is fixedly communicated with the end face of the filter cartridge cover (10), and the other mounting ring (114) is fixedly communicated with the side of the temporary storage cylinder (7).

5. An efficient preparation device for exosome production according to claim 4, characterized in that: A composite processing part (15) is fixedly sleeved on the outer side face of one of the conducting curved pipes (113). The power component (14) drives the engaged composite processing part (15) to rotate. The composite processing part (15) drives the enrichment and collection component (11) to rotate. The composite processing part (15) includes a driven gear II (151) and a convex ring (152). The driven gear II (151) is fixedly sleeved on the outer side of the conducting curved pipe (113). The driven gear II (151) is meshed and connected with the driving gear II (144). The convex ring (152) is fixedly connected to the side face of the driven gear II (151).

6. The highly efficient preparation device for exosome production according to claim 1, characterized in that: The filter element (3) is communicated with the outlet end on the side of the cultivation cylinder (2). The suction end of the peristaltic pump (4) is communicated with the filter element (3). The conveying pipeline assembly includes an elastic diversion pipe fitting (5) and a conducting pipe (6). The upper end of the conducting pipe (6) is fixedly communicated with the temporary storage cylinder (7). The lower end of the conducting pipe (6) is communicated with the elastic diversion pipe fitting (5). The outlet end of the peristaltic pump (4) is communicated with the elastic diversion pipe fitting (5). A liquid accumulation recovery part (18) is arranged below the left end of the elastic diversion pipe fitting (5). A reciprocating pushing component (17) is arranged above the elastic diversion pipe fitting (5). The composite processing part (15) intermittently pushes the reciprocating pushing component (17).

7. The high-efficiency preparation device for exosome production according to claim 6, wherein: The elastic diversion pipe fitting (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 of the elastic connecting pipes (52) is fixedly connected to the conducting pipe (6), and the other elastic connecting pipe (52) is fixedly connected to the outlet end of the peristaltic pump (4).

8. An efficient preparation device for exosome production according to claim 7, characterized in that: The reciprocating pushing assembly (17) includes a linkage member (171), a movable rod (172), and a second spring (173). The upper end of the movable rod (172) is fixedly connected to the linkage member (171). The lower end of the movable rod (172) is movably sleeved in the fitting groove at 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 fitting groove at the top of the assembly plate (1). One end of the linkage member (171) is located above the left end of the pipe body (51), and the other end is located at the bottom of the convex ring (152) and contacts the convex ring (152).

9. The high-efficiency preparation device for exosome production according to claim 8, wherein: The liquid accumulation recovery part (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 communicated with the connecting sleeve (182). The sealing plug (183) is threadedly sleeved in the connecting sleeve (182). The storage cylinder (181) is threadedly sleeved at the lower end of the second pipe (185). The first pipe (184) is fixedly communicated with the bottom of the elastic connecting pipe (52) near the left end of the pipe body (51).

10. The highly efficient preparation device for exosome production according to claim 1, wherein: Both the filter cylinder body (8) and the temporary storage cylinder (7) are fixed to the top of the assembly plate (1) by bottom support rods. The cultivation cylinder (2) and the peristaltic pump (4) are both fixedly installed on the top of the assembly plate (1).

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