Traditional Chinese medicine exosome extraction device

Abstract

The utility model belongs to the field of biological medicine, and relates to a traditional Chinese medicine exosome extraction device, which comprises: a container with a hollow interior and a top provided with an opening for placing raw materials; the microporous filtering membrane is arranged in the container; the stirrer is arranged in the container and is positioned above the microporous filtering membrane; the at least two centrifugal chambers are arranged in the container and located below the microporous filtering membrane, and the centrifugal chambers are arranged from small to large according to the diameters from top to bottom; the centrifugal chambers are annular and are arranged around the same rotating shaft; at least two storage chambers; a waste liquid collection chamber; and pumping devices are arranged in the pumping pipelines, and the pumping pipelines are connected with the centrifugal chambers and the storage chamber, the adjacent centrifugal chambers or the centrifugal chambers and the waste liquid collecting chamber. By arranging the stirrer, the microporous filter membrane, the multi-stage centrifugal chamber and the multi-stage pumping pipeline, the purpose of step-by-step separation is achieved, the extraction speed of the traditional Chinese medicine exosome can be increased, wall breaking and centrifugation are integrated, time is saved, efficiency is improved, and large-scale preparation is achieved.

Description

Technical Field

[0001] The present application relates to the field of biomedicine technology, and in particular to a device for extracting exosomes from traditional Chinese medicine. Background Art

[0002] Exosomes are extracellular vesicles with a diameter of about 30-150 nm that are released from the cell after the fusion of intracellular multivesicular bodies with the cell membrane. Almost all cells can secrete them. Plant-derived exosomes, especially Chinese herbal exosomes, have gradually entered the field of vision of researchers due to their wide range of sources, natural pharmacological activity, low immunogenicity, stable physical and chemical properties, and rich circulation information. The study of Chinese herbal exosomes has not only greatly broadened the range of exosomes, but also provided a new idea for Chinese herbal medicine treatment.

[0003] At present, most Chinese herbal medicines use ultracentrifugation to extract exosomes. Although ultracentrifugation has always been recognized as the gold standard in the industry, the pretreatment process (including cell wall breaking and multiple low-speed centrifugation) is cumbersome, time-consuming and labor-intensive. Utility Model Content

[0004] In view of this, the embodiment of the present application provides a Chinese medicine exosome extraction device to solve the problem of complicated operation of Chinese medicine exosome extraction in the background technology.

[0005] In a first aspect, the present application provides a Chinese medicine exosome extraction device, comprising:

[0006] A container, wherein the interior of the container is hollow and the top has an opening for placing the raw material;

[0007] A microporous filtration membrane is disposed in the container;

[0008] an agitator, disposed in the container and above the microporous filtration membrane;

[0009] At least two centrifugal chambers, which are arranged in the container and below the microporous filtration membrane, and are arranged from top to bottom in terms of diameter from small to large; the centrifugal chambers are all annular and are arranged around the same rotation axis;

[0010] at least two storage chambers disposed in the container and below the microporous filtration membrane;

[0011] Waste liquid collection room;

[0012] A plurality of pumping pipelines are provided with pumping devices in the pumping pipelines, and the pumping pipelines connect the centrifugal chamber and the storage chamber, the adjacent centrifugal chamber, or the centrifugal chamber and the waste liquid collection chamber.

[0013] In conjunction with the first aspect of the present application, in an optional embodiment, the centrifugal chamber includes a primary centrifugal chamber, a secondary centrifugal chamber, a tertiary centrifugal chamber and a quaternary centrifugal chamber arranged from small to large in diameter;

[0014] The storage room includes a primary storage room, a secondary storage room, a tertiary storage room and a quaternary storage room;

[0015] The pumping pipeline includes a primary pipeline, a secondary pipeline, a tertiary pipeline, a quaternary pipeline and a quinary pipeline;

[0016] The primary storage chamber is connected to the primary centrifugal chamber through the primary pipeline; the secondary pipeline connects the secondary centrifugal chamber, the secondary storage chamber and the primary centrifugal chamber; the tertiary pipeline connects the tertiary centrifugal chamber, the tertiary storage chamber and the secondary centrifugal chamber; the quaternary pipeline connects the quaternary centrifugal chamber, the quaternary storage chamber and the tertiary centrifugal chamber; the quaternary centrifugal chamber is connected to the waste liquid collection chamber through the fifth pipeline.

[0017] In combination with the first aspect of the present application, in an optional embodiment, a guide plate is provided in the container, and the guide plate is located below the microporous filtration membrane, and is used to guide the traditional Chinese medicine homogenate processed by the microporous filtration membrane into the primary storage chamber.

[0018] In combination with the first aspect of the present application, in an optional embodiment, the diameter of the first-stage centrifugal chamber is D, the diameter of the second-stage centrifugal chamber is 2D, the diameter of the third-stage centrifugal chamber is 5D, and the diameter of the fourth-stage centrifugal chamber is 50D.

[0019] In conjunction with the first aspect of the present application, in an optional embodiment, the pore size of the microporous filtration membrane is 0.75 μm to 0.85 μm.

[0020] In conjunction with the first aspect of the present application, in an optional implementation manner, the primary pipeline includes a first hose and a first peristaltic pump connected to the first hose;

[0021] The secondary pipeline includes a second hose, a third hose, a second peristaltic pump and a third peristaltic pump; the second hose connects the secondary storage chamber and the primary centrifugal chamber, and the second peristaltic pump is connected to the second hose; the third hose connects the secondary storage chamber and the secondary centrifugal chamber, and the third peristaltic pump is connected to the third hose;

[0022] The tertiary pipeline includes a fourth hose, a fifth hose, a fourth peristaltic pump and a fifth peristaltic pump; the fourth hose connects the tertiary storage chamber and the secondary centrifugal chamber, and the fourth peristaltic pump is connected to the fourth hose; the fifth hose connects the tertiary storage chamber and the tertiary centrifugal chamber, and the fifth peristaltic pump is connected to the fifth hose;

[0023] The four-stage pipeline includes a sixth hose, a seventh hose, a sixth peristaltic pump and a seventh peristaltic pump; the sixth hose connects the four-stage storage chamber and the three-stage centrifugal chamber, and the sixth peristaltic pump is connected to the sixth hose; the seventh hose connects the four-stage storage chamber and the four-stage centrifugal chamber, and the seventh peristaltic pump is connected to the seventh hose;

[0024] The five-stage pipeline includes an eighth hose and an eighth peristaltic pump connected to the eighth hose.

[0025] In combination with the first aspect of the present application, in an optional embodiment, a stirring chamber is formed between the top of the container and the microporous filtration membrane, the stirrer is located in the stirring chamber, and a vacuum pump is also provided in the stirring chamber.

[0026] In combination with the first aspect of the present application, in an optional implementation manner, a sealing cover is installed at the opening.

[0027] In combination with the first aspect of the present application, in an optional implementation manner, the primary pipeline, the secondary pipeline, the tertiary pipeline, the quaternary pipeline and the quintuple pipeline are all arranged in two groups with the rotation axis as the axis of symmetry.

[0028] In combination with the first aspect of the present application, in an optional embodiment, it also includes a temperature controller, wherein the temperature controller is used to adjust the temperature inside the container.

[0029] The Chinese medicine exosome extraction device provided in the embodiment of the present application achieves the purpose of step-by-step separation by providing an agitator, a microporous filtration membrane, a multi-stage centrifugal chamber and a multi-stage pumping pipeline, which can accelerate the speed of Chinese medicine exosome extraction, integrate wall breaking and centrifugation, save time, improve efficiency, and realize large-scale preparation.

[0030] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become apparent from the description below, or will be learned through the practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0032] Figure 1 This is a schematic diagram of the structure of the Chinese medicine exosome extraction device provided in Example 1 of the present application;

[0033] Figure 2 A schematic diagram of the cross-sectional structure (with the symmetrical half omitted) of the Chinese medicine exosome extraction device provided in Example 1 of the present application;

[0034] Figure 3 This is a schematic diagram of the cross-sectional structure (the symmetrical half is omitted) of the Chinese medicine exosome extraction device provided in Example 2 of the present application.

[0035] The reference numerals in the figure are:

[0036] 1. Mixing chamber;

[0037] 2. Agitator;

[0038] 3. Microporous filtration membrane;

[0039] 4. Container;

[0040] 5. Guide plate;

[0041] 10. Primary centrifugal chamber;

[0042] 11. Primary pipeline;

[0043] 12. Primary storage room;

[0044] 20. Secondary centrifugal chamber;

[0045] 21. Secondary pipeline;

[0046] 22. Secondary storage room;

[0047] 30. Three-stage centrifugal chamber;

[0048] 31. Tertiary pipeline;

[0049] 32. Level 3 storage room;

[0050] 40. Four-stage centrifugal chamber;

[0051] 41. Four-stage pipeline;

[0052] 42. Level 4 storage room;

[0053] 51. Five-stage pipeline;

[0054] 52. Waste liquid collection room. DETAILED DESCRIPTION

[0055] The exemplary embodiments disclosed in the present application will be described in more detail below with reference to the accompanying drawings. Although the exemplary embodiments of the present application are shown in the accompanying drawings, it should be understood that the present application can be implemented in various forms and should not be limited by the specific embodiments described herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present application and to fully convey the scope disclosed in the present application to those skilled in the art.

[0056] In the following description, a large number of specific details are given to provide a more thorough understanding of the present application. However, it is obvious to those skilled in the art that the present application can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present application, some technical features known in the art are not described; that is, all features of the actual embodiments are not described here, and well-known functions and structures are not described in detail.

[0057] In the drawings, the sizes of layers, regions, elements and their relative sizes may be exaggerated for clarity. The same reference numerals denote the same elements throughout.

[0058] Spatial relationship terms such as "under", "below", "below", "under", "above", "above", etc., may be used here for convenience of description to describe the relationship between an element or feature shown in the figure and other elements or features. It should be understood that in addition to the orientation shown in the figure, the spatial relationship terms are intended to include different orientations of the device in use and operation. For example, if the device in the accompanying drawings is turned over, then the elements or features described as "under other elements" or "under it" or "under it" will be oriented as "on" other elements or features. Therefore, the exemplary terms "under" and "under" may include both upper and lower orientations. The device can be oriented otherwise (rotated 90 degrees or other orientations) and the spatial descriptors used herein are interpreted accordingly.

[0059] The purpose of the terms used herein is only to describe specific embodiments and is not intended to be limiting of the present application. When used herein, the singular forms "one", "an" and "said / the" are also intended to include plural forms, unless the context clearly indicates otherwise. It should also be understood that the terms "consisting of" and / or "comprising", when used in this specification, determine the presence of the features, integers, steps, operations, elements and / or parts, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, parts and / or groups. When used herein, the term "and / or" includes any and all combinations of the relevant listed items.

[0060] In order to thoroughly understand the present application, detailed steps and detailed structures will be presented in the following description to illustrate the technical solution of the present application. The preferred embodiments of the present application are described in detail below, but in addition to these detailed descriptions, the present application may also have other implementation methods.

[0061] In the prior art, the method of extracting exosomes from traditional Chinese medicine generally uses several test tubes and a high-speed rotating test tube rack. The test tubes filled with raw materials are placed on both sides of the test tube rack, and the driving mechanism is started to drive the test tube ultracentrifugation. In this process, in order to ensure that the test tube rack can run smoothly, each test tube must be strictly weighed to ensure that the weight of the test tubes in the symmetrical position of the test tube rack is consistent to prevent rotation imbalance. In addition, the pretreatment process (including wall breaking and multiple low-speed centrifugation) is also time-consuming, so the preparation of the test tube is relatively cumbersome, the loading capacity is not very high, and the separation efficiency is average. Example 1

[0062] This embodiment provides a Chinese medicine exosome extraction device, such as Figure 1-Figure 2 As shown, including:

[0063] A container 4, wherein the container 4 is hollow inside and has an opening at the top for placing the raw materials;

[0064] A microporous filtration membrane 3 is arranged in the container 4;

[0065] A stirrer 2 is disposed in the container 4 and located above the microporous filtration membrane 3;

[0066] At least two centrifugal chambers, which are arranged in the container 4 and below the microporous filtration membrane 3, and are arranged from small to large in diameter from top to bottom; the centrifugal chambers are all annular and are arranged around the same rotation axis;

[0067] at least two storage chambers, disposed in the container 4 and below the microporous filtration membrane 3;

[0068] Waste liquid collection chamber 52;

[0069] A plurality of pumping pipelines, wherein a pumping device is arranged in the pumping pipeline, and the pumping pipeline connects the centrifugal chamber and the storage chamber, the adjacent centrifugal chamber, or the centrifugal chamber and the waste liquid collection chamber 52.

[0070] The Chinese medicine exosome extraction device provided in this embodiment, the storage chamber is used to store the Chinese medicine homogenate treated by the microporous filter membrane 3 or the centrifugal chamber, and the waste liquid collection chamber 52 is used to collect the waste liquid in the Chinese medicine homogenate treated by the centrifugal chamber at the bottom. There are at least two centrifugal chambers, and they are divided into multiple levels from top to bottom. The function of the pumping pipeline is to pump the extracted liquid processed in the upper centrifugal chamber to the corresponding storage chamber, and then pump it from the storage chamber to the lower centrifugal chamber, and so on, the multi-stage centrifugation process is completed in sequence one by one until the final Chinese medicine exosomes are obtained. Among them, the function of the storage chamber is to cache the extracted liquid processed by the upper centrifugal chamber. Because the centrifugation time of each layer of the centrifugal chamber is generally different, there is a need for a waiting time in the process, so a storage chamber corresponding to each centrifugal chamber is set. The function of the waste liquid collection chamber 52 is to collect the waste liquid separated by the last centrifugal process. The function of the microporous filter membrane 3 is to remove the larger Chinese medicine particles from the stirred Chinese medicine raw material to achieve preliminary filtration.

[0071] The working principle of different centrifugal chambers of the Chinese medicine exosome extraction device provided in this embodiment is: since all the centrifugal chambers are arranged around the rotation axis, all the centrifugal chambers can rotate around the rotation axis at the same angular velocity. According to the centrifugal force formula, when the centrifugal force is at the same angular velocity, the diameter and the centrifugal force are proportional. Therefore, when the centrifugal chambers of the extraction device rotate synchronously, multiple centrifugal chambers with different centrifugal forces are provided. There is no mutual interference between the centrifugal chambers, and the multi-stage centrifugal processes can be carried out simultaneously with high efficiency.

[0072] Optionally, the Chinese medicine exosome extraction device of this embodiment is as follows Figure 2 As shown, the centrifugal chamber includes a first-stage centrifugal chamber 10, a second-stage centrifugal chamber 20, a third-stage centrifugal chamber 30 and a fourth-stage centrifugal chamber 40 arranged from small to large in diameter;

[0073] The storage chamber includes a primary storage chamber 12, a secondary storage chamber 22, a tertiary storage chamber 32 and a quaternary storage chamber 42;

[0074] The pumping pipeline includes a primary pipeline 11, a secondary pipeline 21, a tertiary pipeline 31, a quaternary pipeline 41 and a quinary pipeline 51;

[0075] The primary storage chamber 12 is connected to the primary centrifugal chamber 10 through the primary pipeline 11; the secondary pipeline 21 connects the secondary centrifugal chamber 20, the secondary storage chamber 22 and the primary centrifugal chamber 10; the tertiary pipeline 31 connects the tertiary centrifugal chamber 30, the tertiary storage chamber 32 and the secondary centrifugal chamber 20; the quaternary pipeline 41 connects the quaternary centrifugal chamber 40, the quaternary storage chamber 42 and the tertiary centrifugal chamber 30; the quaternary centrifugal chamber 40 is connected to the waste liquid collection chamber through the fifth pipeline 51.

[0076] In this optional solution, a total of four stages of centrifugation are used, and four corresponding storage chambers are set up, which provide four levels of centrifugal force in sequence, so that the raw materials can be added at one time to directly separate the Chinese medicine exosomes.

[0077] Optionally, the Chinese medicine exosome extraction device of this embodiment is as follows Figure 2 As shown, a guide plate 5 is provided in the container 4, and the guide plate 5 is located below the microporous filtration membrane 3, and is used to guide the Chinese medicine homogenate treated by the microporous filtration membrane 3 to the primary storage chamber 12. Specifically, in this embodiment, the primary storage chamber 12 is located outside the secondary storage chamber 22, and the primary centrifugal chamber 10 is located inside the secondary storage chamber 22. In order to better utilize the space, the Chinese medicine homogenate treated by the microporous filtration membrane 3 is guided to the relatively outer space through the guide plate 5, so as to facilitate the collection of the Chinese medicine homogenate in the primary storage chamber 12.

[0078] Optionally, in the Chinese medicine exosome extraction device of this embodiment, the diameter of the first-stage centrifugal chamber 10 is D, the diameter of the second-stage centrifugal chamber 20 is 2D, the diameter of the third-stage centrifugal chamber 30 is 5D, and the diameter of the fourth-stage centrifugal chamber 40 is 50D. Under this setting, the centrifugal force of the first three centrifugal chambers is gradually increased, and the centrifugal force of the last stage is greatly increased to separate the Chinese medicine exosomes. Specifically, the centrifugal force of the first-stage centrifugal chamber 10 is controlled to be 2000g, the centrifugal force of the second-stage centrifugal chamber 20 is 4000g, the centrifugal force of the third-stage centrifugal chamber 30 is 10000g, and the centrifugal force of the fourth-stage centrifugal chamber 40 is 100000g, D represents a constant, and g is the acceleration of gravity.

[0079] Optionally, in the Chinese medicine exosome extraction device of this embodiment, the pore size of the microporous filter membrane 3 is 0.75 μm-0.85 μm, which is mainly used to remove larger Chinese medicine particles.

[0080] Optionally, the Chinese medicine exosome extraction device of this embodiment is as follows Figure 2 As shown, the primary pipeline 11 includes a first hose and a first peristaltic pump connected to the first hose;

[0081] The secondary pipeline 21 includes a second hose, a third hose, a second peristaltic pump and a third peristaltic pump; the second hose connects the secondary storage chamber 22 and the primary centrifugal chamber 10, and the second peristaltic pump is connected to the second hose; the third hose connects the secondary storage chamber 22 and the secondary centrifugal chamber 20, and the third peristaltic pump is connected to the third hose;

[0082] The tertiary pipeline 31 includes a fourth hose, a fifth hose, a fourth peristaltic pump and a fifth peristaltic pump; the fourth hose connects the tertiary storage chamber 32 and the secondary centrifugal chamber 20, and the fourth peristaltic pump is connected to the fourth hose; the fifth hose connects the tertiary storage chamber 32 and the tertiary centrifugal chamber 30, and the fifth peristaltic pump is connected to the fifth hose;

[0083] The quaternary pipeline 41 includes a sixth hose, a seventh hose, a sixth peristaltic pump and a seventh peristaltic pump; the sixth hose connects the quaternary storage chamber 42 and the tertiary centrifugal chamber 30, and the sixth peristaltic pump is connected to the sixth hose; the seventh hose connects the quaternary storage chamber 42 and the quaternary centrifugal chamber 40, and the seventh peristaltic pump is connected to the seventh hose;

[0084] The five-stage pipeline 51 includes an eighth hose and an eighth peristaltic pump connected to the eighth hose.

[0085] In this optional solution, when the upper centrifugal chamber transports the extract to the lower centrifugal chamber, the extract must first be transported to the storage chamber, and then transported from the storage chamber to the lower centrifugal chamber. This setting can empty the extract processed by the upper centrifugal chamber as quickly as possible without waiting for the lower centrifugal chamber to complete the current centrifugation task. Usually, the lower centrifugation process takes longer, so it is very necessary to set up a storage chamber. In this solution, since the secondary pipeline 21, the tertiary pipeline 31, and the quaternary pipeline 41 all include two independent hoses and two independent peristaltic pumps, the extraction liquid is highly independent in transportation, will not cause conflicts in transportation, and is easy to operate.

[0086] Optionally, in the Chinese medicine exosome extraction device of this embodiment, a stirring chamber 1 is formed between the top of the container 4 and the microporous filter membrane 3, the stirrer 2 is located in the stirring chamber 1, and a vacuum pump is also provided in the stirring chamber 1. Under the action of the vacuum pump, the Chinese medicine homogenate produced in the stirring chamber 1 can pass through the microporous filter membrane 3 and enter the primary storage chamber 12.

[0087] Optionally, in the Chinese medicine exosome extraction device of this embodiment, a sealing cover is installed at the opening to seal the stirring chamber 1.

[0088] Optionally, the Chinese medicine exosome extraction device of this embodiment is as follows Figure 1 As shown, the primary pipeline 11, the secondary pipeline 21, the tertiary pipeline 31, the quaternary pipeline 41 and the quintuple pipeline 51 are all arranged in two groups with the rotation axis as the symmetry axis. This arrangement can improve the efficiency of liquid transportation, or make the two groups of pipelines serve as backup for each other.

[0089] Optionally, the Chinese medicine exosome extraction device of this embodiment further includes a temperature controller, which is used to adjust the temperature inside the container 4 to achieve low-temperature extraction and avoid exosome degradation. The temperature controller can be a compressor or other device.

[0090] The Chinese medicine exosome extraction device of this embodiment takes the white peony root exosome extraction as an example, and the operation steps when using it are as follows:

[0091] Step 1: Take fresh white peony root slices, mix PBS buffer with white peony root and pour into the top opening of container 4, start the stirrer 2, start juicing, and the stirring process is 5 minutes;

[0092] Step 2: After the stirring is completed, the white peony root homogenate is filtered through a microporous filter membrane 3 under the action of a vacuum pump, and the filtering process is 5 min;

[0093] Step 3: After the filtration is completed, all white peony samples are transferred to the primary storage chamber 12 and buffered for 3 minutes; at this time, the stirring and filtration of the next batch of samples can be continued;

[0094] Step 4: The valve of the primary pipeline 11 is opened, and the white peony root sample in the primary storage chamber 12 is transferred to the primary centrifugal chamber 10 by the action of the first peristaltic pump, and the buffering procedure is 3 min;

[0095] Step 5: The valve of the primary pipeline 11 is closed, and the primary centrifugal chamber 10 starts the centrifugal program, and the primary centrifugal program lasts for 20 minutes;

[0096] Step 6: After the first-stage centrifugation is completed, the valve at the second storage chamber 22 end of the second pipeline 21 is opened, and the white peony supernatant in the first centrifugal chamber 10 is transferred to the second storage chamber 22 under the action of the second peristaltic pump, and the valve at the second centrifugal chamber 20 end of the second pipeline 21 is opened, and the white peony supernatant in the second storage chamber 22 is transferred to the second centrifugal chamber 20 under the action of the third peristaltic pump. After the transfer is completed, the valve is closed, and the entire buffering procedure is 3 min;

[0097] Step 7: The secondary centrifugal chamber 20 starts the centrifugation process, and the secondary centrifugation process is 30 minutes; at this time, the primary centrifugation of the next batch of samples can be continued;

[0098] Step 8: After the secondary centrifugation is completed, the valve at the end of the tertiary storage chamber 32 of the tertiary pipeline 31 is opened, and the white peony supernatant in the secondary centrifugal chamber 20 is transferred to the tertiary storage chamber 32 under the action of the fourth peristaltic pump, and the valve at the end of the tertiary centrifugal chamber 30 of the tertiary pipeline 31 is opened, and the white peony supernatant in the tertiary storage chamber 32 is transferred to the tertiary centrifugal chamber 30 under the action of the fifth peristaltic pump. After the transfer is completed, the valve is closed, and the entire buffering procedure is 3 min;

[0099] Step 9: Start the centrifugation process in the three-stage centrifuge chamber. The three-stage centrifugation process lasts for 60 minutes. At this time, the second-stage centrifugation of the next batch of samples can be continued.

[0100] Step 10: After the third-stage centrifugation is completed, the valve at the end of the fourth-stage storage chamber 42 of the fourth-stage pipeline 41 is opened, and the white peony supernatant in the third-stage centrifugal chamber 30 is transferred to the fourth-stage storage chamber 42 under the action of the sixth peristaltic pump, and the valve at the end of the fourth-stage centrifugal chamber 40 of the fourth-stage pipeline 41 is opened, and the white peony supernatant in the fourth-stage storage chamber 42 is transferred to the fourth-stage centrifugal chamber 40 under the action of the seventh peristaltic pump. After the transfer is completed, the valve is closed, and the entire buffering procedure takes 3 minutes;

[0101] Step 11: The four-stage centrifugal chamber 40 starts the centrifugation process, and the four-stage centrifugation process is 70 minutes; at this time, the three-stage centrifugation of the next batch of samples can be continued;

[0102] Step 12: After the fourth-stage centrifugation is completed, the valve of the fifth-stage pipeline 51 is opened, and the supernatant of the fourth-stage centrifugal chamber 40 is transferred to the waste liquid collection chamber 52 under the action of the eighth peristaltic pump. The precipitate obtained in the fourth-stage centrifugal chamber 40 is the white peony exosomes. Example 2

[0103] like Figure 3 As shown, this embodiment provides a pipeline structure different from the solution of embodiment 1. In this solution, since the secondary pipeline 21, the tertiary pipeline 31, and the quaternary pipeline 41 are all three-way hoses, the three pipe openings of the three-way hose are respectively connected to the centrifugal chamber located relatively in the upper layer, the centrifugal chamber located relatively in the lower layer, and the corresponding storage chamber. Valves and peristaltic pumps are set on each branch of the three-way hose, and the transfer of the extract between the centrifugal chamber and the storage chamber can also be achieved. Compared with the solution of embodiment 1, the occupied volume of the pipeline is small, but the control of the valve and the peristaltic pump is more complicated.

[0104] It should be understood that the above embodiments are exemplary and are not intended to include all possible implementations included in the claims. Various modifications and changes may be made on the basis of the above embodiments without departing from the scope of the present disclosure. Similarly, the various technical features of the above embodiments may be arbitrarily combined to form other embodiments of the present application that may not be explicitly described. Therefore, the above embodiments only express several implementations of the present application and do not limit the scope of protection of the patent of this application.

Claims

1. A Chinese medicine exosome extraction device, characterized in that: include: A container (4), wherein the interior of the container (4) is hollow and the top has an opening for placing the raw materials; A microporous filtration membrane (3) is arranged in the container (4); A stirrer (2) is arranged in the container (4) and located above the microporous filtration membrane (3); at least two centrifugal chambers, the centrifugal chambers being arranged in the container (4) and below the microporous filtration membrane (3), and arranged from top to bottom in terms of diameter from small to large; the centrifugal chambers are all annular and are arranged around the same rotation axis; At least two storage chambers, arranged in the container (4) and located below the microporous filtration membrane (3); a waste liquid collection chamber (52); A plurality of pumping pipelines, each of which is provided with a pumping device, and each of which connects the centrifugal chamber and the storage chamber, an adjacent centrifugal chamber, or the centrifugal chamber and the waste liquid collection chamber (52).

2. The Chinese medicine exosome extraction device according to claim 1, characterized in that: The centrifugal chamber comprises a first-stage centrifugal chamber (10), a second-stage centrifugal chamber (20), a third-stage centrifugal chamber (30) and a fourth-stage centrifugal chamber (40) arranged in ascending order of diameter; The storage chamber comprises a primary storage chamber (12), a secondary storage chamber (22), a tertiary storage chamber (32) and a quaternary storage chamber (42); The pumping pipeline comprises a primary pipeline (11), a secondary pipeline (21), a tertiary pipeline (31), a quaternary pipeline (41) and a quinary pipeline (51); The primary storage chamber (12) is connected to the primary centrifugal chamber (10) via the primary pipeline (11); the secondary pipeline (21) connects the secondary centrifugal chamber (20), the secondary storage chamber (22) and the primary centrifugal chamber (10); the tertiary pipeline (31) connects the tertiary centrifugal chamber (30), the tertiary storage chamber (32) and the secondary centrifugal chamber (20); the quaternary pipeline (41) connects the quaternary centrifugal chamber (40), the quaternary storage chamber (42) and the tertiary centrifugal chamber (30); and the quaternary centrifugal chamber (40) is connected to the waste liquid collection chamber via the quintuple pipeline (51).

3. The Chinese medicine exosome extraction device according to claim 2, characterized in that: A guide plate (5) is provided in the container (4), and the guide plate (5) is located below the microporous filtration membrane (3) and is used to guide the Chinese medicine homogenate liquid processed by the microporous filtration membrane (3) into the primary storage chamber (12).

4. The Chinese medicine exosome extraction device according to claim 2, characterized in that: The diameter of the first-stage centrifugal chamber (10) is D, the diameter of the second-stage centrifugal chamber (20) is 2D, the diameter of the third-stage centrifugal chamber (30) is 5D, and the diameter of the fourth-stage centrifugal chamber (40) is 50D.

5. The Chinese medicine exosome extraction device according to claim 4, characterized in that: The pore size of the microporous filtration membrane (3) is 0.75 μm to 0.85 μm.

6. The Chinese medicine exosome extraction device according to any one of claims 2 to 5, characterized in that: The primary pipeline (11) comprises a first hose and a first peristaltic pump connected to the first hose; The secondary pipeline (21) comprises a second hose, a third hose, a second peristaltic pump and a third peristaltic pump; the second hose connects the secondary storage chamber (22) and the primary centrifugal chamber (10), and the second peristaltic pump is connected to the second hose; the third hose connects the secondary storage chamber (22) and the secondary centrifugal chamber (20), and the third peristaltic pump is connected to the third hose; The tertiary pipeline (31) comprises a fourth hose, a fifth hose, a fourth peristaltic pump and a fifth peristaltic pump; the fourth hose connects the tertiary storage chamber (32) and the secondary centrifugal chamber (20), and the fourth peristaltic pump is connected to the fourth hose; the fifth hose connects the tertiary storage chamber (32) and the tertiary centrifugal chamber (30), and the fifth peristaltic pump is connected to the fifth hose; The quaternary pipeline (41) comprises a sixth hose, a seventh hose, a sixth peristaltic pump and a seventh peristaltic pump; the sixth hose connects the quaternary storage chamber (42) and the tertiary centrifugal chamber (30), and the sixth peristaltic pump is connected to the sixth hose; the seventh hose connects the quaternary storage chamber (42) and the quaternary centrifugal chamber (40), and the seventh peristaltic pump is connected to the seventh hose; The five-stage pipeline (51) comprises an eighth hose and an eighth peristaltic pump connected to the eighth hose.

7. The Chinese medicine exosome extraction device according to any one of claims 1 to 5, characterized in that: A stirring chamber (1) is formed between the top of the container (4) and the microporous filtration membrane (3), the stirrer (2) is located in the stirring chamber (1), and a vacuum pump is also provided in the stirring chamber (1).

8. The Chinese medicine exosome extraction device according to claim 7, characterized in that: A sealing cover is installed at the opening.

9. The Chinese medicine exosome extraction device according to any one of claims 2 to 5, characterized in that: The primary pipeline (11), the secondary pipeline (21), the tertiary pipeline (31), the quaternary pipeline (41) and the quintuple pipeline (51) are all arranged in two groups with the rotation axis as a symmetry axis.

10. The Chinese medicine exosome extraction device according to any one of claims 1 to 5, characterized in that: It also includes a temperature controller, which is used to adjust the temperature inside the container (4).

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