Stem cell culture supernatant exosome real-time interception culture bottle

Abstract

The utility model discloses a stem cell culture supernatant exosome real-time interception culture bottle which comprises an interception culture bottle body and a bottle cap, the bottle cap is buckled at the position of an upper port of the interception culture bottle body, a liquid inlet pipe and an exosome sampling pipe are inserted into the bottle cap, one side of the exosome sampling pipe is connected with a negative pressure collecting assembly, and the other side of the exosome sampling pipe is connected with a liquid outlet pipe. The lower end of the bottle body of the interception culture bottle is connected with a liquid outlet pipe, and one end of the liquid outlet pipe is communicated with the culture cavity. The utility model relates to the technical field of stem cell culture, a detachable split type interception assembly is integrally installed in the bottle body, and after the bottle cap is removed, the split type interception assembly can be integrally taken out; the inner supporting net rack and the filter membrane cylinder are detached, so that the filter membrane cylinder is quickly replaced, the filter membrane replacement convenience is greatly improved, the exposed end of the exosome sampling pipe is connected with the negative pressure collecting assembly, real-time closure of the exosome can be achieved, the structure is simple, and operation is convenient.

Description

Technical Field

[0001] This utility model relates to the field of stem cell culture technology, specifically to a culture flask for real-time retention of exosomes in stem cell culture supernatant. Background Technology

[0002] Stem cell-derived exosomes, present in stem cell supernatants, are nanovesicles secreted by stem cells, carrying various functional proteins and genetic information, such as nucleic acids, proteins, and lipids. The real-time exosome trapping culture flask is a novel experimental device designed to optimize exosome collection processes. Its core function is to directly capture secreted exosomes during cell culture, avoiding damage to the vesicle structure caused by traditional separation methods. In existing technologies, trapping culture flasks typically use a filter membrane to control the directional flow of exosome-containing supernatant for continuous interception and separation. However, current structures rely heavily on the pre-installed filter membrane's lifespan (approximately 10 cycles), which is prone to clogging and requires frequent replacement, making replacement inconvenient. Therefore, this invention addresses these issues through in-depth research. Utility Model Content

[0003] To address the shortcomings of existing technologies, this invention provides a culture flask for real-time retention of exosomes in stem cell culture supernatant, thus solving the problems mentioned in the background art.

[0004] To achieve the above objectives, this utility model provides the following technical solution: a stem cell culture supernatant exosome real-time interception culture bottle, comprising an interception culture bottle body and a cap, wherein the cap is fastened to the port end of the interception culture bottle body, an inlet tube and an exosome sampling tube are inserted into the cap, a negative pressure collection component is connected to one side of the exosome sampling tube, an outlet tube is connected to the lower end of the interception culture bottle body, one end of the outlet tube is connected to the culture chamber, one end of the culture chamber is connected to the inlet tube via a peristaltic pump, and an assembly ring is provided at the opening at the upper end of the interception culture bottle body, wherein a split-type interception component is inserted into the assembly ring;

[0005] The split-type interception assembly includes a fixing ring, a basket-type filter cylinder, a filter membrane cylinder, and an inner support frame. The upper end face of the mounting ring has several limiting notches. The basket-type filter cylinder is disposed on the lower end of the fixing ring. The outer end face of the fixing ring is provided with limiting protrusions that match the limiting notches. The filter membrane cylinder is disposed inside the basket-type filter cylinder. The inner support frame is inserted into the filter membrane cylinder. A limiting screw sleeve is screwed into the fixing ring. The lower end of the limiting screw sleeve is in contact with the upper end face of the inner support frame.

[0006] A rubber sealing gasket is provided on the top of the aforementioned assembly ring.

[0007] The inner ring of the aforementioned fixed ring is provided with a threaded groove, and the outer ring of the limiting screw sleeve is provided with an external thread that matches the threaded groove. Two fixed crossbars are symmetrically arranged inside the limiting screw sleeve.

[0008] The aforementioned negative pressure collection assembly includes a support, a collection chamber, a piston, a pull rod, and a drive control component. The collection chamber is snapped onto the support, and one end of the collection chamber is connected to an exosome sampling tube via a connecting pipe. The piston is inserted into the collection chamber and slides in a seal with the inner annular surface of the collection chamber. The pull rod is fixedly connected to one end of the piston, and the drive control component is mounted on the support with its moving end connected to the pull rod.

[0009] The aforementioned drive control components include a lead screw module and a connecting seat. The lead screw module is mounted on a support, and the connecting seat is installed on the moving end of the lead screw module and engages with the pull rod.

[0010] The aforementioned pull rod has a retaining plate at one end, and the connecting seat has a retaining groove corresponding to the retaining plate position. Beneficial effects

[0011] This invention provides a real-time exosome retention culture bottle for stem cell culture supernatant. It offers the following advantages: This real-time exosome retention culture bottle improves upon existing retention culture bottles by integrating a detachable, split-type retention component within the bottle body. After removing the cap, the split-type retention component can be removed entirely, allowing for the disassembly of the internal support frame and filter membrane cartridge, thus enabling rapid replacement of the filter membrane cartridge and significantly improving the convenience of filter replacement. The exposed end of the exosome sampling tube is connected to a negative pressure collection component, enabling real-time retention of exosomes. The structure is simple and the operation is convenient. The stem cell supernatant inside the retention culture bottle and in the culture chamber circulates using gravity and a peristaltic pump, effectively ensuring the continuity of exosome retention operations. Attached Figure Description

[0012] Figure 1 This is a schematic diagram of the main structure of the stem cell culture supernatant exosome real-time retention culture flask described in this utility model.

[0013] Figure 2 This utility model Figure 1 A partially enlarged structural diagram.

[0014] Figure 3 This is an isometric structural diagram of the basket-type filter cylinder described in this utility model.

[0015] Figure 4 This is an isometric structural diagram of the internal support grid frame described in this utility model.

[0016] Figure 5 This is an isometric structural diagram of the limiting screw sleeve described in this utility model.

[0017] In the diagram: 1. Body of the culture flask; 2. Cap; 3. Inlet tube; 4. Exosome sampling tube; 5. Outlet tube; 6. Culture chamber; 7. Peristaltic pump; 8. Assembly ring; 9. Fixing ring; 10. Basket-type filter cartridge; 11. Filter membrane cartridge; 12. Internal support frame; 13. Limiting screw sleeve; 14. Rubber sealing gasket; 15. Fixing crossbar; 16. Support; 17. Collection chamber; 18. Piston; 19. Pull rod; 20. Lead screw module; 21. Connecting seat; 22. Clamping plate. Detailed Implementation

[0018] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0019] Example: Refer to the appendix of the instruction manual Figure 1-5As can be seen, this application specifically designs a stem cell culture supernatant exosome real-time interception culture bottle, including an interception culture bottle body 1 and a bottle cap 2. The bottle cap 2 is fastened to the upper port of the interception culture bottle body 1. An inlet tube 3 and an exosome sampling tube 4 are inserted into the bottle cap 2. A negative pressure collection component is connected to one side of the exosome sampling tube 4. An outlet tube 5 is connected to the lower end of the interception culture bottle body 1. One end of the outlet tube 5 is connected to a culture chamber 6. One end of the culture chamber 6 is connected to the inlet tube 3 through a peristaltic pump 7. An assembly ring 8 is provided at the upper opening of the interception culture bottle body 1. A split interception component is inserted into the assembly ring 8. The split interception component includes a fixing The assembly ring 8 comprises a fixed ring 9, a basket-type filter cartridge 10, a filter membrane cartridge 11, and an inner support mesh frame 12. The upper surface of the assembly ring 8 has several limiting notches. The basket-type filter cartridge 10 is mounted on the lower end of the fixed ring 9. The outer end face of the fixed ring 9 has limiting protrusions that match the limiting notches. The filter membrane cartridge 11 is placed inside the basket-type filter cartridge 10. The inner support mesh frame 12 is inserted into the filter membrane cartridge 11. A limiting screw sleeve 13 is screwed into the fixed ring 9, with the lower end of the limiting screw sleeve 13 abutting against the upper surface of the inner support mesh frame 12. A rubber sealing gasket 14 is provided at the top of the assembly ring 8. The inner surface of the fixed ring 9 has a threaded groove, and the outer surface of the limiting screw sleeve 13 has a matching threaded groove. The external thread of the limiting screw sleeve 13 has two symmetrically arranged fixing crossbars 15. This improves upon the existing intercepting culture bottle body 1 and cap 2 by integrating a detachable, split-type interception component within the bottle body. The exosome-containing supernatant (diameter 30–150 nm) secreted by stem cells passes through the filter membrane cartridge 11 (pore size 30–150 nm). Exosomes are trapped on the filter membrane surface, while small molecule metabolites and culture medium components freely pass through the filter membrane and return to the culture chamber 6 for recycling. When the filter membrane cartridge 11 needs to be replaced, first remove the cap 2, then pull up the crossbar inside the limiting screw sleeve 13 to remove and disassemble the split-type interception component as a whole. The limiting screw sleeve 13 is used to pull out the inner support mesh frame 12 and the filter membrane tube 11. The new filter membrane tube 11 is then fitted onto the inner support mesh frame 12 and reinstalled into the basket-type filter mesh tube. The limiting screw sleeve 13 is tightened, and the split-type interception component is reinstalled into the bottle body, thereby realizing the rapid replacement of the filter membrane tube 11 and greatly improving the convenience of filter membrane replacement. The exposed end of the exosome sampling tube 4 is connected to a negative pressure collection component, which can realize the real-time interception of exosomes. The structure is simple and the operation is convenient. The stem cell supernatant inside the culture bottle body 1 and the culture chamber 6 is circulated by gravity and peristaltic pump 7, which can effectively ensure the continuity of exosome interception operation.

[0020] In a preferred embodiment, the negative pressure collection assembly includes a support 16, a collection chamber 17, a piston 18, a pull rod 19, and a drive control component. The collection chamber 17 is snapped onto the support 16, and one end of the collection chamber 17 is connected to the exosome sampling tube 4 via a connecting pipe. The piston 18 is inserted into the collection chamber 17 and slides and seals against the inner annular surface of the collection chamber 17. The pull rod 19 is fixedly connected to one end of the piston 18. The drive control component is mounted on the support 16, and its moving end is connected to the pull rod 19. The drive control component includes a lead screw module 20 and a connecting seat 21. The lead screw module 20 is mounted on the support 16, and the connecting seat 21 is mounted on the lead screw. The movable end of module 20 is engaged with pull rod 19. One end of pull rod 19 is provided with a locking plate 22. The connecting seat 21 is provided with a locking groove corresponding to the locking plate 22. Using screw module 20 as power source, the locking plate 22 is pulled by support 16, thereby adjusting the position of pull rod 19. The movement of pull rod 19 drives piston 18 to move. The piston 18 moves away from the bottle body, thereby generating negative pressure in collection chamber 17. Under the action of negative pressure, the exosomes intercepted in the culture bottle are sucked into collection chamber 17 through exosome sampling tube 4. The operation is simple, continuous and can be automatically controlled, thus realizing the real-time interception of exosomes.

[0021] It should be noted that, in this document, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, the phrase "comprising an element defined as..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0022] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A stem cell culture supernatant exosome real-time retention culture flask, comprising a flask body and a cap, wherein the cap is fastened to the port of the flask body, characterized in that, The bottle cap is fitted with an inlet tube and an exosome sampling tube. A negative pressure collection assembly is connected to one side of the exosome sampling tube. An outlet tube is connected to the lower end of the throttling culture bottle. One end of the outlet tube is connected to the culture chamber. One end of the culture chamber is connected to the inlet tube via a peristaltic pump. An assembly ring is provided at the opening at the upper end of the throttling culture bottle. A split-type interception assembly is inserted into the assembly ring. The split-type interception assembly includes a fixing ring, a basket-type filter cylinder, a filter membrane cylinder, and an inner support frame. The upper end face of the mounting ring has several limiting notches. The basket-type filter cylinder is disposed on the lower end of the fixing ring. The outer end face of the fixing ring is provided with limiting protrusions that match the limiting notches. The filter membrane cylinder is disposed inside the basket-type filter cylinder. The inner support frame is inserted into the filter membrane cylinder. A limiting screw sleeve is screwed into the fixing ring. The lower end of the limiting screw sleeve is in contact with the upper end face of the inner support frame.

2. The stem cell culture supernatant exosome real-time retention culture flask according to claim 1, characterized in that, A rubber sealing gasket is provided on the top of the assembly ring.

3. The stem cell culture supernatant exosome real-time interception culture flask of claim 1, wherein, The inner ring of the fixed ring is provided with a threaded groove, and the outer ring of the limiting screw sleeve is provided with an external thread that matches the threaded groove. Two fixed crossbars are symmetrically arranged inside the limiting screw sleeve.

4. The stem cell culture supernatant exosome real-time interception culture flask of claim 1, wherein, The negative pressure collection assembly includes a support, a collection chamber, a piston, a pull rod, and a drive control component. The collection chamber is snapped onto the support, and one end of the collection chamber is connected to an exosome sampling tube via a connecting pipe. The piston is inserted into the collection chamber and slides in a seal with the inner annular surface of the collection chamber. The pull rod is fixedly connected to one end of the piston. The drive control component is mounted on the support, and its moving end is connected to the pull rod.

5. The stem cell culture supernatant exosome real-time interception culture flask of claim 4, wherein, The drive control unit includes a lead screw module and a connecting seat. The lead screw module is mounted on a support, and the connecting seat is mounted on the moving end of the lead screw module and engages with the pull rod.

6. The stem cell culture supernatant exosome real-time interception culture flask of claim 5, wherein, One end of the pull rod is provided with a retaining plate, and the connecting seat is provided with a retaining groove corresponding to the position of the retaining plate.

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