A stem cell exosome extraction and storage device

Abstract

The application belongs to the technical field of exosome extraction devices, and discloses a stem cell exosome extraction and storage device, wherein a base frame is provided with a fixing assembly for fixing a test tube; the base frame is provided with a blow and beat assembly, a swing assembly and a driving assembly; the blow and beat assembly comprises a syringe, the outlet of the syringe is connected with an inlet and outlet assembly, the inlet and outlet assembly is provided with a plurality of liquid outlets along the circumference of the test tube, and the liquid outlet direction of the liquid outlets is parallel to the tangent direction of the test tube; the piston rod of the syringe is connected with the driving assembly; the swing assembly comprises a swing piece rotatably connected with the base frame, one end of the swing piece is located in the test tube, and the other end of the swing piece is connected with the driving assembly; the driving assembly drives the swing piece to swing on a vertical plane, and drives the piston rod of the syringe to reciprocate along the axial direction of the syringe. The application solves the problems that the current artificial knocking is easy to cause the rupture and structural damage of exosome vesicles, and the blow and beat of a pipette gun is not uniform and the precipitation is not completely dispersed.

Description

Technical Field

[0001] This invention relates to the field of exosome extraction device technology, specifically a stem cell exosome extraction and storage device. Background Technology

[0002] Stem cells possess the biological characteristics of self-replication and multi-lineage differentiation, and have broad application prospects in regenerative medicine, clinical diagnosis and treatment, and other fields. Their sources mainly include peripheral blood stem cells, bone marrow stem cells, and stem cells from other tissues. After extraction and purification using specialized stem cell separation solutions, the stem cells needed for clinical treatment are obtained. However, stem cells are difficult to preserve and transport, limiting their further research and application. Research has found that vesicles secreted by stem cells, namely exosomes, have similar biological functions to stem cells. Currently, the research and application of stem cell exosomes has become a hot topic in the field of stem cell research. Ultracentrifugation has become one of the mainstream methods for separating stem cell exosomes due to its mature technology, large sample processing capacity, and considerable product purity and yield. This method removes cells, debris, and large particles through multi-stage centrifugation, ultimately enriching the exosome precipitate. After ultracentrifugation, the enriched exosome precipitate gathers at the bottom and corners of the centrifuge tube. Resuspension is necessary to evenly disperse the vesicles in buffer solution before subsequent processing.

[0003] Currently, the common method to loosen the precipitate is to manually tap the bottom of the tube. However, the mechanical impact and vibration from tapping can easily damage the exosome structure, causing exosome vesicle rupture, structural damage, and a significant decrease in activity. Another method is to use a pipette to mix the precipitate along the tube wall, but this is limited by the direction of the liquid flow and the range of action. Areas such as the center of the cavity and the bottom corners are prone to forming mixing blind zones, resulting in uneven mixing and incomplete dispersion of the precipitate. Furthermore, the operator's technique, experience, and force control vary greatly, and the force of blowing or tapping can be inconsistent. This not only leads to uneven mixing effects for a single batch of samples but also causes significant differences in the integrity and uniformity of exosomes between different batches. Summary of the Invention

[0004] The present invention aims to provide a stem cell exosome extraction and storage device, which solves the problems of exosome vesicle rupture and structural damage caused by manual tapping, as well as uneven agitation and incomplete precipitation dispersion caused by pipetting.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] A stem cell exosome extracting and storing device, comprising a centrifugal device and a resuspension auxiliary device, wherein the resuspension auxiliary device comprises a base frame, and the base frame is provided with a fixing assembly for fixing a test tube; a pipetting component, a swinging component and a driving component are arranged on the base frame above the test tube; the pipetting component comprises a syringe, an outlet of the syringe is connected with a liquid inlet and outlet component, the liquid inlet and outlet component is provided with a plurality of liquid outlets along the circumferential direction of the test tube, and the liquid outlet direction of the liquid outlets is parallel to the tangential direction of the test tube; a piston rod of the syringe is connected with the driving component; the swinging component comprises a swinging member rotationally connected with the base frame, one end of the swinging member is located in the test tube, and the other end of the swinging member is connected with the driving component; the driving component drives the swinging member to swing in a vertical plane, and simultaneously drives the piston rod of the syringe to reciprocate along the axial direction of the syringe.

[0007] Further, the driving component comprises a rotating shaft rotationally connected with the base frame, an eccentric cam and a rocking lever are fixedly connected to the rotating shaft, a free end of the rocking lever is rotationally connected with a connecting rod, and a free end of the connecting rod is rotationally connected with the swinging member; the base frame is slidably connected with a connecting frame along the axial direction of the syringe, an inner wall of the connecting frame is in sliding abutment with the eccentric cam, and the connecting frame is connected with the piston rod of the syringe; the base frame is provided with a motor for driving the rotating shaft to rotate.

[0008] Further, the connecting frame is connected with a convex-shaped sliding rail, the base frame is provided with a convex-shaped sliding groove, and the sliding rail is slidably connected in the sliding groove.

[0009] Further, the connecting frame and the piston rod of the syringe are detachably connected by a buckle, a bolt or a pin.

[0010] Further, the liquid inlet and outlet component comprises a first connecting pipe, one end of the first connecting pipe is communicated with the syringe, the other end of the first connecting pipe is connected with an annular pipeline, the annular pipeline is connected with a plurality of liquid outlet pipes along the circumferential direction, and the liquid outlets are arranged on side walls of the liquid outlet pipes.

[0011] Further, the syringe is connected with a second connecting pipe, and one end of the second connecting pipe away from the syringe is located in the test tube; both the first connecting pipe and the second connecting pipe are provided with one-way valves, the one-way valve of the first connecting pipe controls the unidirectional flow of liquid from the inside of the syringe to the inside of the test tube, and the one-way valve of the second connecting pipe controls the unidirectional flow of liquid from the inside of the test tube to the inside of the syringe.

[0012] Further, the swinging member comprises a swinging rod, and one end of the swinging rod located in the test tube is connected with a small ball.

[0013] Furthermore, the fixing component includes a support component and two sets of clamping components, the clamping components being used to clamp the sidewall of the test tube, and the support component being used to support the bottom of the test tube.

[0014] Furthermore, the clamping assembly includes a sleeve connected to the base frame, a slide rod slidably connected inside the sleeve, and a clamping pad connected to the free end of the slide rod; a spring is provided outside the slide rod, one end of the spring is connected to the support pad, and the other end of the spring is connected to the sleeve.

[0015] Furthermore, the support assembly includes a screw rod that is threadedly connected to the base frame, one end of the screw rod being connected to a support plate, and the other end of the screw rod being connected to a knob handle.

[0016] The principles and beneficial effects of the technical solution are as follows:

[0017] 1. This invention provides a stem cell exosome extraction and storage device. A centrifuge device is used to remove cells, debris, and large particles in stages. After centrifugation, the enriched exosome precipitate gathers at the bottom and corner of the sidewall of the centrifuge tube. A resuspension operation is then performed to uniformly disperse the vesicles in a buffer solution before further processing. A resuspension aid device ensures uniform dispersion of the vesicles in the buffer solution. Specifically, the resuspension aid device includes a base frame with a fixing component to support and fix the test tube, preventing it from shaking or falling during the mixing and dispersion of exosomes. A blowing component, a oscillating component, and a driving component are located above the test tube on the base frame. The blowing component includes a syringe connected to an inlet / outlet component, which has several outlets along the circumference of the test tube, with the outlet direction parallel to the tangent of the test tube. The oscillating component includes an oscillating element rotatably connected to the base frame, with one end of the oscillating element located inside the test tube. The driving component is connected to a piston rod and the oscillating element.

[0018] When using this device, after ultracentrifugation and the accumulation of exosome precipitate at the bottom of the tube, add an appropriate volume of sterile buffer solution to the test tube. Then place the test tube on the base frame and fix it in place with the fixing component to firmly support the test tube and prevent it from shaking or slipping during subsequent mixing. After the test tube is positioned, start the drive component. The drive component synchronously links the piston rod and the swinging component of the syringe to simultaneously perform tangential fluid circulation and central swinging disturbance. The two actions work together to achieve gentle and comprehensive dispersion of exosome precipitate.

[0019] Specifically, when the piston rod slides upward, it draws the buffer solution inside the test tube into the syringe cavity. When the piston rod slides downward, it pushes the liquid from the syringe to the outlets arranged around the circumference of the test tube. The liquid is sprayed into the test tube along a tangential direction parallel to the test tube wall, forming a tangential circulation. Unlike traditional pipettes that directly blow liquid onto the precipitate below the surface, this device uses tangential fluid to spread along the tube wall. Relying on the drag force of the liquid wall and boundary layer penetration, it gradually wets and loosens the exosome precipitate at the bottom of the tube, avoiding concentrated high-pressure impacts. The overall shear force of the fluid is significantly reduced, effectively preventing direct, strong flow from tearing the exosome's lipid membrane and damaging vesicle bioactivity. Simultaneously, multiple outlets are evenly distributed around the circumference, allowing the circulating fluid to cover the entire sidewall of the test tube, overcoming the limitation of manual single-point blowing which only affects a localized area of ​​the tube wall and reducing mixing dead zones in the sidewall region. Furthermore, compared to manual vibration and tapping of the test tube, this tangential fluid mixing method avoids instantaneous, violent mechanical impacts and high-frequency vibrations, effectively preventing damage to the fragile exosome lipid membrane caused by tube wall compression and vibration stress, thus reducing vesicle breakage and loss at the source.

[0020] Simultaneously, the drive component synchronously drives the oscillating component to swing back and forth in the vertical plane inside the test tube, using the hinge point as a fulcrum. The end of the oscillating component extends into the center of the test tube cavity, continuously agitating the top layer of liquid. The oscillating component directly acts on the middle, and the reciprocating swing can drive the central static liquid to generate a weak up-down and oblique turbulent flow, which carries the exosome aggregates deposited at the bottom center and deep corners of the test tube that have not been entrained by the circulation flow into the circulating liquid flow. This effectively avoids the mixing blind zone that is common in traditional resuspension methods, allowing the liquid in the entire tube to participate in mixing synchronously, resulting in more uniform and thorough precipitation dispersion.

[0021] The drive component synchronously links the syringe piston rod and the oscillating component. The tangential fluid blowing speed generated by the reciprocating push and pull of the piston rod matches the reciprocating oscillation speed of the oscillating component, avoiding speed mismatches such as rapid fluid circulation and insufficient oscillation, or excessively high oscillation frequency and lagging fluid circulation. The two mixing forces are superimposed in real time, resulting in a stable and uniform composite mixing effect. Furthermore, it eliminates the need for manual pipetting and repeated blowing, or manual tapping of the tube bottom, avoiding inconsistent mixing effects within the same batch of samples and poor stability between different batches caused by manual operation. The disturbance intensity, circulation flow rate, and oscillation amplitude are uniformly controllable throughout the process, significantly improving the experimental repeatability and batch consistency of exosome preparation. Simultaneously, the combination of low-impact tangential fluid and reciprocating oscillation, without violent vibration, high-pressure direct jets, or high-shear turbulence, allows for thorough dispersion and precipitation of exosomes in a gentle physical environment, maximizing the protection of the structural integrity and biological function of the fragile nanoscale vesicles, effectively improving the exosome extraction recovery rate and product quality.

[0022] 2. The present invention provides a stem cell exosome extraction and storage device, the driving component of which includes a rotating shaft, an eccentric cam and a rocker arm connected to the rotating shaft, a connecting rod rotatably connected to the rocker arm, and a rotatably connected oscillating component; a connecting frame is slidably connected to the base frame, the upper and lower inner walls of the connecting frame slidingly abutting against the eccentric cam, and the connecting frame is connected to the piston rod of the syringe; in use, the motor drives the rotating shaft to rotate, and the eccentric cam and rocker arm synchronously move in a circular motion with the rotating shaft; during the rotation of the eccentric cam, it uses its own eccentric contour to alternately push against the upper and lower inner walls of the connecting frame, driving the connecting frame to slide back and forth along the vertical height direction, thereby driving the piston rod to push and pull, realizing the circulation of liquid along the tangent direction of the test tube; the same rotating shaft synchronously drives the rocker arm to rotate, and the rocker arm is transmitted through the connecting rod to pull the oscillating component located on the upper layer of liquid in the test tube to swing back and forth in the vertical plane inside the test tube.

[0023] In other words, the eccentric cam and rocker arm are coaxially arranged, allowing the tangential fluid blowing action and the upper oscillating component's stirring action to start and stop synchronously. The tangential circulation downwards along the pipe wall draws in the bottom sediment and transports it upwards to the upper liquid layer. The lateral vortex generated by the oscillation of the oscillating component extends downwards, enhancing the vertical convection of the liquid inside the pipe and indirectly driving the bottom sediment to continuously participate in the circulation and mixing. The rotating shaft is a unified power source, with the rotation cycle of the eccentric cam synchronized with the oscillation cycle of the rocker arm. The motor speed determines the piston rod liquid circulation flow rate and the oscillation frequency of the oscillating component, ensuring that the two speeds are always matched and preventing misalignment of fluid delivery and upper layer stirring rhythm. In addition, the connecting frame bidirectionally cancels the lateral component force of the eccentric cam, and the piston rod only bears the axial force, resulting in smooth sliding and preventing wear and leakage. The height of the connecting frame allows for flexible limitation of the piston rod's stroke, adapting to different syringe ranges. At the same time, the connecting frame acts as a guide and limiter, preventing the piston rod from deviating and jamming. The "convex" shaped slide rail and "convex" shaped groove guide and limit the movement of the connecting frame. In addition, the swinging component includes a swing rod, one end of which is connected to a small ball inside the test tube. The small ball can be made of medical-grade silicone to reduce motion resistance and transmission load.

[0024] 3. The present invention provides a stem cell exosome extraction and storage device. The inlet and outlet assembly includes a connecting tube, one end of which is connected to a syringe, and the other end of which is connected to an annular pipe. Several outlet pipes are connected circumferentially along the annular pipe, with the outlet located on the side wall of the outlet pipe. In use, the liquid inside the test tube can be discharged and circulated back through the outlet, thereby achieving tangential mixing of the exosome precipitate at the bottom. The annular pipe, combined with the multiple circumferentially arranged outlet pipes, enables synchronous discharge from the entire inner wall of the test tube, resulting in more uniform loosening of the exosome precipitate deposited at corners. The annular pipe also functions as a liquid storage and pressure stabilizer, and provides uniform flow distribution. In addition, a second connecting tube is added to the syringe, with its end extending into the middle of the test tube as a reflux channel. One-way valves are installed on both connecting tubes. The one-way valve on connecting tube one restricts liquid flow only from the syringe to the test tube, while the one-way valve on connecting tube two restricts liquid flow only from the test tube back to the syringe. This creates a dual-channel structure with a central reflux inlet and a circumferential outlet. This central reflux and circumferential outlet layout creates a stable, directional circulating flow field inside the test tube. The fluid around the perimeter flows downwards to trap sediment, while the central reflux transports particles upwards, significantly improving liquid replacement efficiency.

[0025] 4. The present invention provides a stem cell exosome extraction and storage device, wherein the fixing component includes a support component and two sets of clamping components. The clamping component includes a sleeve, a sliding rod slidably connected inside the sleeve, a clamping pad connected to the sliding rod, and a spring sleeved on the sliding rod. The support component includes a screw rod threadedly connected to a base frame, and a support plate connected to the screw rod. In use, first rotate the bottom screw to move the circular support plate downward, leaving sufficient space for placement. At this time, the two sets of clamping pads are in an inward-folding state under the spring's rebound. Spread the two sets of clamping pads, and the slide bar retracts into the sleeve to compress the spring. Place the test tube between the two clamping pads and adjust the height. The spring applies a counter-force, and the clamping pads on both sides automatically adhere to and hug the side wall of the test tube, completing the lateral pre-clamping. Then, rotate the screw forward to drive the support plate to rise vertically until the top surface of the support plate is in contact with the bottom of the test tube, supporting the test tube from below, completing the dual positioning of lateral clamping and bottom lifting. At the same time, the height of the support plate can be finely adjusted by the screw to accurately match the working position of the upper annular liquid outlet pipe and the upper swing ball.

[0026] In other words, the sleeve provides horizontal guidance and constraint to the slide rod, preventing it from tilting or jamming during extension and retraction, ensuring that the clamping pads on both sides are always aligned, and allowing the slide rod to smoothly transmit the spring force, resulting in uniform force on the clamping pads without bias. The spring sleeved on the outside of the slide rod acts as a flexible clamping power source. When the test tube is placed in, it is squeezed and compressed, and the spring continuously holds the test tube with its gentle rebound force. This allows it to adapt to test tubes of different outer diameters without the need to change clamps. The screw continuously drives the support plate to move vertically up and down using a self-locking threaded pair. Lowering the support plate can reserve space for placing the tube, share the force on the lateral clamping pads, prevent the test tube from tilting or swaying, and ensure that the relative position of the test tube, the upper pipeline, and the oscillating ball remains constant, resulting in a stable mixing flow field. After adjustment, the screw has a self-locking capability, preventing the support plate from sliding down on its own during operation. This adapts to test tubes of different lengths and liquid levels, and the adjustment operation is simple and effortless. In addition, the design of the support components and the two sets of clamping components does not obstruct the sides of the test tube, making it easy to observe the dispersion and mixing process. Attached Figure Description

[0027] Figure 1 This is a top view of a stem cell exosome extraction and storage device according to the present invention;

[0028] Figure 2 for Figure 1 Sectional view of AA;

[0029] Figure 3 for Figure 2 Enlarged view of point D;

[0030] Figure 4 for Figure 2 Sectional view of BB;

[0031] Figure 5 This is a schematic diagram of the structure of a stem cell exosome extraction and storage device according to the present invention;

[0032] Figure 6 This is a front view of a stem cell exosome extraction and storage device according to the present invention;

[0033] Figure 7 for Figure 6 A sectional view of CC.

[0034] The names of the corresponding labels in the attached diagram are:

[0035] 1. Base frame; 2. Screw; 3. Knob handle; 4. Support plate; 5. Test tube; 6. Sleeve; 7. Slide rod; 8. Spring; 9. Clamping pad; 10. Guide block; 11. Swinging component; 12. Swinging rod; 13. Small ball; 14. Syringe; 15. Piston rod; 16. Rotating shaft; 17. Eccentric cam; 18. Connecting frame; 19. Slide rail; 20. Slide groove; 21. Rocker arm; 22. Connecting rod; 23. Annular pipe; 24. Connecting pipe one; 25. Connecting pipe two; 26. Discharge pipe; 27. Motor. Detailed Implementation

[0036] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments:

[0037] like Figures 1 to 7 As shown, a stem cell exosome extraction and storage device includes a centrifugation device for removing cells, debris, and large particles in stages; a resuspension auxiliary device includes a base frame 1, which can be fixed to a platform by bolts or welding. The base frame 1 is provided with a fixing component for fixing test tubes 5; the fixing component includes a support component and two sets of clamping components. The clamping components are used to clamp the sidewalls of the test tubes 5, and the support component is used to support the bottom of the test tubes 5. In some embodiments, the clamping component includes a sleeve 6 connected to the base frame 1. A slide rod 7 is slidably connected inside the sleeve 6. Specifically, a guide block 10 is connected inside the sleeve 6. The slide rod 7 is provided with a guide groove, which is slidably connected to the outside of the guide block 10. The free end of the slide rod 7 is connected to a clamping pad 9, which can be made of rubber. A spring 8 is sleeved on the slide rod 7. One end of the spring 8 is connected to the support pad, and the other end of the spring 8 is connected to the sleeve 6. In addition, the support component includes a screw 2 connected to the base frame 1 by threads. One end of the screw 2 is connected to a support plate 4, and the other end of the screw 2 is connected to a knob handle 3.

[0038] A blowing assembly, a swinging assembly, and a driving assembly are provided on the base frame 1 above the test tube 5. The blowing assembly includes a syringe 14, the piston rod 15 of which is connected to the driving assembly. The outlet of the syringe 14 is connected to an inlet / outlet assembly, which has several outlets along the circumference of the test tube 5, and the outlet direction is parallel to the tangential direction of the test tube 5. In some embodiments, the inlet / outlet assembly includes a connecting tube 24, one end of which is connected to the syringe 14, and the other end of which is connected to an annular pipe 2. 3. The annular pipe 23 is connected to several outlet pipes 26 along the circumference, and the outlet is located on the side wall of the outlet pipe 26. In addition, in order to separate the inlet channel and the outlet channel, the syringe 14 is also connected to a connecting pipe 25. The end of the connecting pipe 25 away from the syringe 14 is located in the test tube 5. Both the connecting pipe 1 24 and the connecting pipe 2 25 are equipped with one-way valves. The one-way valve of the connecting pipe 1 24 controls the liquid to flow unidirectionally from the syringe 14 to the test tube 5, and the one-way valve of the connecting pipe 2 25 controls the liquid to flow unidirectionally from the test tube 5 to the syringe 14.

[0039] The swing assembly includes a swing member 11 rotatably connected to the base frame 1. One end of the swing member 11 is located inside the test tube 5, and the other end of the swing member 11 is connected to the drive assembly. In some embodiments, the swing member 11 includes a swing rod 12. One end of the swing rod 12 located inside the test tube 5 is connected to a small ball 13. The small ball 13 may be made of medical silicone.

[0040] While the drive assembly drives the oscillating member 11 to oscillate in the vertical plane, it also drives the piston rod 15 of the syringe 14 to slide back and forth along the axial direction of the syringe 14. The drive assembly includes a rotating shaft 16 rotatably connected to the base frame 1. The base frame 1 is provided with a motor 27 for driving the rotating shaft 16 to rotate. The rotating shaft 16 is fixedly connected to an eccentric cam 17 and a rocker arm 21. The free end of the rocker arm 21 is rotatably connected to a connecting rod 22, and the free end of the connecting rod 22 is rotatably connected to the oscillating member 11. The base frame 1 is slidably connected to a connecting frame 18 along the axial direction of the syringe 14. The upper and lower inner walls of the connecting frame 18 slide against the eccentric cam 17. Specifically, the connecting frame 18 is connected to a U-shaped slide rail 19, and the base frame 1 is provided with a U-shaped slide groove 20. The slide rail 19 is slidably connected in the slide groove 20. The connecting frame 18 is connected to the piston rod 15 of the syringe 14. In some embodiments, the connecting frame 18 and the piston rod 15 of the syringe 14 are detachably connected by a buckle, bolt, or pin. Furthermore, centrifuge devices are existing technology and will not be described in detail here.

[0041] The specific implementation process is as follows:

[0042] First, adjust the height of test tube 5 relative to the swinging component 11 and the liquid outlet tube 26, and then fix it with the fixing component; start the motor 27, and the rotating shaft 16 synchronously drives the eccentric cam 17 and the rocker arm 21 to rotate. The eccentric cam 17, in conjunction with the connecting frame 18, converts the circular motion into vertical reciprocating linear motion, continuously pushing and pulling the piston rod 15 of the syringe 14 to complete the reciprocating suction and discharge action; during the liquid pushing stage of the piston rod 15, the liquid is synchronously sprayed out from the liquid outlets on the side walls of the multiple circumferential liquid outlet tubes 26 along the inner wall of the test tube 5, relying on the drag force of the fluid adhering to the wall to gently flush the exosomes deposited at the bottom of the tube layer by layer; during the liquid suction stage of the piston rod 15, the liquid flows back to the syringe 14. The dual-channel structure of liquid outlet at all four sides and liquid return in the middle forms a stable directional circulating flow field, continuously transporting the bottom exosomes to the upper layer of liquid. Simultaneously, the rocker arm 21 drives the upper swing arm 12 to swing back and forth, causing the central static liquid to generate a weak up-and-down and oblique turbulent flow, which carries the exosome aggregates deposited at the bottom center and deep corners of the test tube 5 that have not been entrained by the circulation flow into the circulating liquid flow. After continuous circulation and mixing until the exosomes inside and outside the tube are completely and evenly dispersed, the motor 27 is turned off, the support plate 4 is lowered to release the bottom support, and the test tube 5 can be removed.

[0043] When the piston rod 15 slides upward, it draws the buffer solution inside the test tube 5 into the cavity of the syringe 14. When the piston rod 15 slides downward, it pushes the liquid from the syringe 14 to the outlet arranged around the test tube 5. The liquid is sprayed into the test tube 5 along a tangential direction parallel to the wall of the test tube 5, forming a tangential circulation. Unlike traditional pipettes that insert directly under the liquid surface to blow away sediment, this device uses tangential fluid to spread along the tube wall. Relying on the drag force of the liquid wall and boundary layer penetration, it gradually wets and loosens the exosome sediment at the bottom of the tube, avoiding concentrated high-pressure impacts. The overall shear force of the fluid is significantly reduced, effectively preventing direct, strong flow from tearing the exosome's lipid membrane and damaging vesicle bioactivity. Simultaneously, multiple outlets are evenly distributed around the circumference, allowing the circulating fluid to cover the entire sidewall of the tube 5, overcoming the limitation of manual single-point blowing which only affects a localized area of ​​the tube wall and reducing mixing dead zones in the sidewall region. Furthermore, compared to manual vibration and tapping of the tube 5, this tangential fluid mixing method avoids instantaneous, violent mechanical impacts and high-frequency vibrations, effectively preventing damage to the fragile exosome lipid membrane due to tube wall compression and vibration stress, thus reducing vesicle breakage and loss at the source.

[0044] Simultaneously, the drive component synchronously drives the oscillating element 11 to swing back and forth in the vertical plane inside the test tube 5, using the hinge point as a fulcrum. During this process, the test tube 5 can also be rotated, allowing the oscillating element 11 to agitate in each vertical plane. The end of the oscillating element 11 extends into the center of the cavity of the test tube 5, continuously agitating the top layer of liquid. The oscillating element 11 directly acts on the middle, and the reciprocating swing can drive the central static liquid to generate a weak up-down and oblique turbulent flow, which carries the exosome aggregates deposited at the bottom center and deep corners of the test tube 5 that have not been entrained by the circulation flow into the circulating liquid flow. This effectively avoids the mixing blind zone that is common in traditional resuspension methods, allowing the liquid in the entire tube to participate in mixing synchronously, resulting in more uniform and thorough precipitation dispersion.

[0045] The driving component synchronously links the piston rod 15 and the oscillating component 11 of the syringe 14. The tangential fluid blowing speed generated by the reciprocating push and pull of the piston rod 15 matches the reciprocating oscillation speed of the oscillating component 11, avoiding speed misalignment issues such as fast fluid circulation and sluggish oscillation, or excessively high oscillation frequency and lagging fluid circulation. The two mixing forces are superimposed in real time, resulting in a stable and uniform composite mixing effect. Furthermore, it eliminates the need for manual hand-held pipette repeated blowing and tapping of the tube bottom, avoiding the inconsistent mixing effects of the same batch of samples and poor stability of different batches of finished products caused by manual operation. The disturbance intensity, circulation flow rate, and oscillation amplitude are uniformly controllable throughout the process, significantly improving the experimental repeatability and batch consistency of exosome preparation. Simultaneously, the combination of tangential low-impact fluid and reciprocating oscillation, without violent vibration, high-pressure direct jets, or high-shear turbulence, allows for thorough dispersion and precipitation of exosomes in a gentle physical environment, maximizing the protection of the structural integrity and biological function of the nanoscale fragile vesicles, effectively improving the exosome extraction recovery rate and product quality.

[0046] The eccentric cam 17 and rocker arm 21 are coaxially arranged, allowing the tangential fluid blowing action and the stirring action of the upper oscillating component 11 to start and stop synchronously. The tangential circulating flow downward along the pipe wall, sucking up the bottom sediment and conveying it upward to the upper liquid layer. The lateral vortex generated by the oscillation of the oscillating component 11 extends downward, enhancing the vertical convection of the liquid inside the pipe and indirectly driving the bottom sediment to continuously participate in the circulation and mixing. The rotating shaft 16 is a unified power source, and the rotation period of the eccentric cam 17 is synchronized with the oscillation period of the rocker arm 21. The speed of the motor 27 determines the liquid circulation flow rate of the piston rod 15 and the oscillation component. The oscillation frequency of the 11 swing rods ensures that the speeds of the two components are always matched, preventing any misalignment in fluid delivery or upper agitation rhythm. Furthermore, the connecting frame 18 bidirectionally cancels the lateral force of the eccentric cam 17, and the piston rod 15 only bears axial force, resulting in smooth sliding and minimal wear and leakage. The height of the connecting frame 18 allows for flexible limitation of the piston rod 15's stroke, adapting to different syringe 14 ranges. One end of the swing rod 12 located inside the test tube 5 is rotatably connected to a small ball 13, which can be made of medical-grade silicone to reduce movement resistance and transmission load.

[0047] The annular pipe 23, combined with multiple circumferentially arranged outlet pipes 26, enables synchronous liquid discharge throughout the inner wall of the test tube 5, resulting in more uniform loosening of exosomes deposited at corners. The annular pipe 23 also functions as a liquid storage and pressure stabilizer, and a uniform flow distribution system. In addition, the syringe 14 is equipped with a second connecting pipe 25 connected to it. Both the first connecting pipe 24 and the second connecting pipe 25 are fitted with one-way valves. The layout of central reflux and peripheral liquid discharge creates a stable directional circulating flow field inside the test tube 5. The fluid around the perimeter draws down the sediment and flows back up in the center, significantly improving the liquid replacement efficiency.

[0048] Sleeve 6 forms a horizontal guiding constraint on slide rod 7, preventing slide rod 7 from tilting or jamming during extension and retraction, ensuring that the clamping pads 9 on both sides are always centered, and slide rod 7 smoothly transmits the elastic force of spring 8, so that the clamping pads 9 are evenly stressed without bias pressure; spring 8, sleeved on the outside of slide rod 7, serves as a flexible clamping power source. When test tube 5 is placed, it is squeezed and compressed, and the spring force continuously holds test tube 5 tightly, which can adapt to test tubes 5 with different outer diameters without the need to change clamps. Screw 2 continuously drives support plate 4 to move vertically up and down by means of self-locking thread pair. Lowering support plate 4 can reserve space for tube placement, share the force on the lateral clamping pads 9, prevent test tube 5 from tilting and shaking, and ensure that the relative position of test tube 5 with the upper pipeline and swing ball 13 is constant, and the mixed flow field is stable; after adjustment, screw 2 has self-locking capability, and support plate 4 will not slide down on its own during operation, adapting to test tubes 5 of different lengths and liquid levels, and the adjustment operation is simple and labor-saving.

[0049] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific technical solutions or characteristics is not elaborated upon here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the technical solutions of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A stem cell exosome extraction and storage device, comprising a centrifugation device and a resuspension auxiliary device, characterized in that, The resuspension auxiliary device comprises a base frame (1), wherein the base frame (1) is provided with a fixing component for fixing a test tube (5); a blowing and beating component, a swinging component and a driving component are arranged on the base frame (1) above the test tube (5); the blowing and beating component comprises a syringe (14), an inlet and outlet liquid component is connected to an outlet of the syringe (14), the inlet and outlet liquid component is provided with a plurality of liquid outlets along the circumferential direction of the test tube (5), and a liquid outlet direction of the liquid outlets is parallel to a tangential direction of the test tube (5); a piston rod (15) of the syringe (14) is connected with the driving component; the swinging component comprises a swinging member (11) rotationally connected with the base frame (1), one end of the swinging member (11) is located in the test tube (5), and the other end of the swinging member (11) is connected with the driving component; the driving component drives the swinging member (11) to swing in a vertical plane and simultaneously drives the piston rod (15) of the syringe (14) to slide back and forth along the axial direction of the syringe (14).

2. The stem cell exosome extraction and storage device according to claim 1, characterized in that, The driving component comprises a rotating shaft (16) rotationally connected with the base frame (1), an eccentric cam (17) and a rocker (21) are fixedly connected to the rotating shaft (16), a free end of the rocker (21) is rotationally connected with a connecting rod (22), and a free end of the connecting rod (22) is rotationally connected with the swinging member (11); a connecting frame (18) is slidably connected to the base frame (1) along the axial direction of the syringe (14), an inner wall of the connecting frame (18) is in sliding abutting contact with the eccentric cam (17), and the connecting frame (18) is connected with the piston rod (15) of the syringe (14); the base frame (1) is provided with a motor (27) for driving the rotating shaft (16) to rotate.

3. The stem cell exosome extraction and storage device according to claim 2, characterized in that, The connecting frame (18) is connected with a convex-shaped sliding rail (19), the base frame (1) is provided with a convex-shaped sliding chute (20), and the sliding rail (19) is slidably connected in the sliding chute (20).

4. The stem cell exosome extraction and storage device according to claim 2, characterized in that, The connecting frame (18) and the piston rod (15) of the syringe (14) are detachably connected through a buckle, a bolt or a pin.

5. The stem cell exosome extraction and storage device according to claim 1, characterized in that, The inlet and outlet liquid component comprises a first connecting pipe (24), one end of the first connecting pipe (24) communicates with the syringe (14), the other end of the first connecting pipe (24) is connected with an annular pipeline (23), a plurality of liquid outlet pipes (26) are connected to the annular pipeline (23) along the circumferential direction, and the liquid outlets are arranged on side walls of the liquid outlet pipes (26).

6. The stem cell exosome extraction and storage device according to claim 5, characterized in that, The syringe (14) is connected with a second connecting pipe (25), one end of the second connecting pipe (25) away from the syringe (14) is located in the test tube (5); both the first connecting pipe (24) and the second connecting pipe (25) are provided with one-way valves, the one-way valve of the first connecting pipe (24) controls liquid to flow unidirectionally from the inside of the syringe (14) into the test tube (5), and the one-way valve of the second connecting pipe (25) controls liquid to flow unidirectionally from the inside of the test tube (5) into the syringe (14).

7. The stem cell exosome extraction and storage device according to claim 1, characterized in that, The swinging component (11) includes a swinging rod (12), one end of which is located inside the test tube (5) and is connected to a small ball (13).

8. The stem cell exosome extraction and storage device according to claim 1, characterized in that, The fixing component includes a support component and two sets of clamping components. The clamping components are used to clamp the side wall of the test tube (5), and the support component is used to support the bottom of the test tube (5).

9. The stem cell exosome extraction and storage device according to claim 8, characterized in that, The clamping assembly includes a sleeve (6) connected to the base frame (1), a slide rod (7) slidably connected inside the sleeve (6), and a clamping pad (9) connected to the free end of the slide rod (7); a spring (8) is provided on the outer sleeve of the slide rod (7), one end of the spring (8) is connected to the support pad, and the other end of the spring (8) is connected to the sleeve (6).

10. The stem cell exosome extraction and storage device according to claim 8, characterized in that, The support assembly includes a screw (2) that is threadedly connected to the base frame (1), one end of the screw (2) is connected to a support plate (4), and the other end of the screw (2) is connected to a knob handle (3).

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