Exosome culture and collection device

Abstract

The utility model discloses an exosome culture and collection device. The exosome culture and collection device comprises a culture assembly, a collection assembly and a purification assembly, according to the exosome collecting device, the culture bottle, the collecting bottle and the centrifugal tube are sequentially connected through the detachable connector, so that the operation of collecting exosomes is changed into semi-closed operation from original open operation, and the pollution risk in the collecting process can be effectively reduced; the third connector is connected with a negative pressure liquid suction pump, the third connector is connected with a negative pressure liquid suction pump, the third connector is connected with a negative pressure liquid suction pump, and the negative pressure liquid suction pump is connected with a negative pressure liquid suction pump. And after the negative pressure liquid suction pump is opened, liquid in the collection bottle can quickly enter the high-speed centrifugal tube through the micro-filtration membrane, so that bacteria can be effectively removed, cell fragments and protein fragments with smaller diameters can be filtered out, and the collection and purification efficiency is improved.

Description

Technical Field

[0001] The utility model relates to the technical field of biological vessels, and in particular to an exosome culture and collection device. Background Art

[0002] Exosomes are small membrane vesicles (30-150 nm) that contain complex RNA and proteins. They contain a variety of biomolecules, including lipids, proteins, nucleic acids (such as RNA and DNA), and carbohydrates. Exosomes are primarily released outside cells through a process called exocytosis and can then be taken up by other cells. They have applications in medical fields such as immune regulation, tumor development, and neurotransmission.

[0003] Currently, when transferring cell fluid containing exosomes from cell culture flasks to collection flasks, laboratories need to manually collect the cell supernatant, transfer it to a centrifuge tube using a pipette, manually filter it through a microfiltration membrane, and then transfer it to a centrifuge tube for ultrahigh-speed centrifugation to collect the exosome suspension. Because the entire process is open, the cell supernatant is exposed to the air during the operation, greatly increasing the risk of sample contamination. Microorganisms, dust, and other contaminants in the air may enter the sample, which not only affects the accuracy of the experimental results but may also cause experimental failure. Utility Model Content

[0004] The purpose of the present invention is to solve the above problems and provide an exosome culture and collection device.

[0005] In order to achieve the above-mentioned purpose, the present invention specifically adopts the following technical solutions, including:

[0006] a culture component, a collection component, and a purification component, wherein both ends of the collection component are detachably connected via a hose;

[0007] The culture component includes a culture bottle, which is used to culture cells; the collection component includes a collection bottle, which is used to collect exosomes; and the purification component includes a centrifuge tube, which is used to centrifuge and purify exosomes;

[0008] The culture bottle and the collection bottle are connected with a connector, and the connector is used to filter impurities.

[0009] As a further description of the above technical solution, the culture flask adopts a multi-layer cell culture device, and the culture flasks are stacked in 1-5 layers.

[0010] As a further description of the above technical solution, connecting tubes are provided at both ends of the top of the collecting bottle, and the connecting tubes are used to connect to the connector.

[0011] As a further description of the above technical solution, the connector includes a first connector, a second connector and a third connector.

[0012] As a further description of the above technical solution, a filter membrane is provided inside the connector.

[0013] As a further description of the above technical solution, the first connector is arranged on one side of the culture bottle body, and the pore size of the filter membrane in the first connector is 90-120 μm, preferably 100 μm.

[0014] As a further description of the above technical solution, the second connector is arranged on one side of the collecting bottle, and the pore size of the filter membrane in the second connector is 30-50 μm, preferably 40 μm.

[0015] As a further description of the above technical solution, the third connector is arranged on the other side of the collecting bottle, and the pore size of the filter membrane in the second connector is 0.20-0.25 μm, preferably 0.22 μm.

[0016] As a further description of the above technical solution, the third connector is connected to a suction pump, which is used to transfer the liquid in the collection bottle to the centrifuge tube.

[0017] As a further description of the above technical solution, a microfiltration membrane is provided in the centrifuge tube, and the pore size of the microfiltration membrane is 100-120 nm, preferably 120 nm.

[0018] The beneficial effects of the utility model are as follows:

[0019] 1. This utility model uses a detachable connector to sequentially connect the culture bottle, collection bottle, and centrifuge tube, so that the operation of collecting exosomes is changed from the original open operation to a semi-closed operation, which can effectively reduce the risk of contamination during the collection process;

[0020] 2. The utility model is provided with three groups of connectors. The pore size of the internal filter membrane gradually decreases. By gradually filtering, the cell supernatant passes through the filter membranes of different pore sizes, and the cell debris and large impurities are filtered out in turn, and then ultra-high-speed centrifugation is performed to purify the exosomes. The third connector is also connected to a negative pressure suction pump. After turning on the negative pressure suction pump, the liquid in the collection bottle can quickly pass through the microfiltration membrane into the high-speed centrifuge tube, which can effectively sterilize and filter out cell fragments and protein fragments with smaller diameters, thereby improving the collection and purification efficiency.

[0021] In order to more clearly illustrate the structural features and functions of the present invention, the present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a structural schematic diagram of the exosome culture and collection device of the present invention.

[0023] Reference numerals:

[0024] 1. Culture component; 11. Culture bottle; 2. Collection component; 21. Collection bottle; 3. Purification component; 31. Centrifuge tube; 4. Hose; 5. Connector; 51. First connector; 52. Second connector; 53. Third connector; 6. Connecting tube; 7. Microfiltration membrane. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention.

[0026] like Figure 1 As shown, in one embodiment, an exosome culture and collection device includes: a culture component 1, a collection component 2 and a purification component 3, and the two ends of the collection component 2 are detachably connected to the collection component 2 and the purification component 3 through a hose 4.

[0027] like Figure 1 As shown, in this embodiment, the culture component 1 includes a culture bottle 11 for culturing cells to secrete exosomes; the collection component 2 includes a collection bottle 21 for collecting cell supernatant containing exosomes; and the purification component 3 includes a centrifuge tube 31, which is specifically used for the centrifugal purification process of exosomes.

[0028] In addition, a connector 5 is provided between the culture bottle 11 and the collection bottle 21 to gradually filter out impurities and ensure the purity of the collected exosome sample.

[0029] Specifically, the culture flask 11 adopts a multi-layer cell culture apparatus, and the culture flask 11 is stacked in 1-5 layers. It is equipped with a full set of accessories, including liquid transfer caps, small-mouth conversion caps, adapters, three-way pipes, silicone tubes / hot-melt tubes, quick connectors, etc. Through the mutual cooperation of different pipes and accessories, aseptic transfer of liquid is achieved, which effectively reduces the risk of cell contamination. To expand the culture scale, it is only necessary to increase the number of stacked layers without taking up too much space; and connecting tubes 6 are provided at both ends of the top of the collection bottle 21, and the connecting tubes 6 are used to connect to the connector 5.

[0030] like Figure 1 As shown, in this embodiment, the connector 5 includes a first connector 51, a second connector 52, and a third connector 53. The first connector 51 is arranged on one side of the culture bottle 11, the second connector 52 is arranged on one side of the collection bottle 21, and the third connector 53 is arranged on the other side of the collection bottle 21. A filter membrane is arranged inside the connector 5.

[0031] Specifically, the pore size of the filter membrane in the first connector 51 is 90-120 μm, and the laboratory uses 100 μm. The pore size of the filter membrane in the second connector 52 is 30-50 μm, and the laboratory uses 40 μm. The pore size of the filter membrane in the second connector 52 is 0.20-0.25 μm, and the laboratory uses 0.22 μm.

[0032] Furthermore, the third connector 53 is connected to a liquid suction pump, which is used to transfer the liquid in the collection bottle 21 into the centrifuge tube 31 .

[0033] like Figure 1 As shown, in this embodiment, a microfiltration membrane 7 is further provided in the centrifuge tube 31. Specifically, the pore size of the microfiltration membrane 7 is 100-120 nm, and 120 nm is used in the laboratory.

[0034] Working principle:

[0035] First, cells are cultured using the culture component 1 to secrete exosomes. The main component is the culture bottle 11. Cells can be cultured in the culture bottle 11. After the cells secrete exosomes, they are collected in a timely manner. The culture bottle 11 is detachably connected to the first connector 51 and contains a 100μm filter membrane.

[0036] Secondly, use the collection component 2 to collect the cell supernatant: open the first connector 51 of the culture bottle 11, and connect the plastic hose 4, the second connector 52 and the receiving bottle in sequence; invert the culture bottle 11, so that the cell supernatant (containing exosomes) first passes through a 100μm filter membrane to filter out cell debris and large impurities, and then passes through the plastic hose 4 through a 40μm filter membrane to filter again, and finally flows into the receiving bottle. If the filter is found to be clogged during filtration, it can be disassembled and replaced with a new connector 5.

[0037] Finally, to remove more impurities and enrich the exosomes for a higher purity, a centrifuge assembly is used for centrifugation. When using the centrifuge assembly, the culture assembly 1 can be removed first, leaving only the receiving assembly: the receiving bottle is connected to the ultracentrifuge tube 31 via the third connector 53. One end of the third connector 53 is connected to a negative pressure liquid pump. Turning on the negative pressure liquid pump allows the liquid in the collection bottle 21 to quickly pass through the 0.22μm filter membrane and enter the high-speed centrifuge tube 31, effectively sterilizing and filtering out smaller cell fragments and protein fragments. After removing the third connector 53, the ultracentrifuge tube 31 containing the 120nm microporous filter membrane is now filled with relatively pure liquid. The high-speed centrifuge tube 31 is then centrifuged to obtain pure exosomes.

[0038] Through the above technical solution, the present application uses a detachable connector 5 to sequentially connect the culture bottle 11, the collection bottle 21 and the centrifuge tube 31, so that the exosome collection operation is changed from the original open operation to a semi-closed operation, significantly reducing the risk of contamination; the pore size of the filter membrane gradually decreases, and through step-by-step filtration, the cell supernatant passes through filter membranes of different pore sizes in sequence to remove cell debris and large impurities, and then ultra-high-speed centrifugation is performed to purify the exosomes; and the second connector 52 is also connected to a negative pressure suction pump. After it is turned on, the liquid in the collection bottle 21 can be quickly transported to the high-speed centrifuge tube 31 through the microfiltration membrane 7, which can effectively sterilize and filter out cell fragments and protein fragments with smaller diameters, thereby improving the collection and purification efficiency.

[0039] The above description of the disclosed embodiments will enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. An exosome culture and collection device, characterized in that: include: A culture component (1), a collection component (2) and a purification component (3), wherein both ends of the collection component (2) are detachably connected to the collection component (2) and the purification component (3) via a hose (4); The culture component (1) includes a culture bottle (11), the culture bottle (11) is used to culture cells, the collection component (2) includes a collection bottle (21), the collection bottle (21) is used to collect exosomes, and the purification component (3) includes a centrifuge tube (31), the centrifuge tube (31) is used to centrifuge and purify the exosomes; The culture bottle (11) and the collection bottle (21) are connected with a connector (5), and the connector (5) is used to filter impurities.

2. The exosome culture and collection device according to claim 1, characterized in that: The culture flask (11) adopts a multi-layer cell culture device, and the culture flask (11) is stacked in 1-5 layers.

3. The exosome culture and collection device according to claim 1, characterized in that: Connecting tubes (6) are provided at both ends of the top of the collecting bottle (21), and the connecting tubes (6) are used to connect to the connector (5).

4. The exosome culture and collection device according to claim 1, characterized in that: The connector (5) comprises a first connector (51), a second connector (52) and a third connector (53).

5. The exosome culture and collection device according to claim 1, characterized in that: A filter membrane is provided inside the connector (5).

6. The exosome culture and collection device according to claim 4, characterized in that: The first connector (51) is arranged on one side of the body of the culture bottle (11), and the pore size of the filter membrane in the first connector (51) is 90-120 μm.

7. The exosome culture and collection device according to claim 4, characterized in that: The second connector (52) is arranged on one side of the collecting bottle (21), and the pore size of the filter membrane in the second connector (52) is 30-50 μm.

8. The exosome culture and collection device according to claim 4, characterized in that: The third connector (53) is arranged on the other side of the collecting bottle (21), and the pore size of the filter membrane in the second connector (52) is 0.20-0.25 μm.

9. The exosome culture and collection device according to claim 4, characterized in that: The third connector (53) is connected to a liquid suction pump, and the liquid suction pump is used to transfer the liquid in the collecting bottle (21) into the centrifuge tube (31).

10. The exosome culture and collection device according to claim 1, characterized in that: A microfiltration membrane (7) is provided in the centrifuge tube (31), and the pore size of the microfiltration membrane (7) is 100-120 nm.

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