CN214552580U - Cell vesicle purification device - Google Patents
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- CN214552580U CN214552580U CN202120316035.1U CN202120316035U CN214552580U CN 214552580 U CN214552580 U CN 214552580U CN 202120316035 U CN202120316035 U CN 202120316035U CN 214552580 U CN214552580 U CN 214552580U
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Abstract
The utility model provides a cell vesicle purification device, including container lid, first container, second container, third container, fourth container and the filtrating collection portion that from top to bottom connects gradually, through first microfiltration membrane intercommunication between first container and the second container, through second microfiltration membrane intercommunication between second container and the third container, through third microfiltration membrane intercommunication between third container and the fourth container, fourth container and filtrating collection portion intercommunication, first microfiltration membrane's aperture is 1-5 mu m, and second microfiltration membrane's aperture is 500-. The utility model discloses a cell vesicle purification device simple structure, easily operation, it is high to the extraction efficiency of cell vesicle.
Description
Technical Field
The utility model relates to a cell vesicle purification field, concretely relates to cell vesicle purification device.
Background
The cytoskeleton is an important structure by which the eukaryotic cell maintains its basic morphology, and the cell can cause the cytoskeleton to be rearranged after being stimulated exogenously or endogenously, so that the cell membrane is locally stressed unevenly, and the abnormally stressed cytoplasm expands outwards, and then the abnormally stressed cytoplasm is randomly coated with part of the cell contents and then released to the outside of the cell in the form of vesicles, so that a special subcellular structure with the diameter of about 0.1-1 μm is formed, and the structure is called as a 'cell vesicle'. The generation method and application of cell vesicles have been reported, for example, chinese patent document CN102302784B discloses that ultraviolet rays are used to irradiate tumor cells to obtain cell vesicles, or chemotherapy drugs serving as active ingredients are applied to tumor cells to cause apoptosis, so as to obtain cell vesicles released by tumor cells, and then the cell vesicles and the chemotherapy drugs are incubated at room temperature to obtain a tumor chemotherapy drug preparation. Has important significance for the extraction and purification treatment of the cell vesicles for practical application.
At present, vesicle extraction and purification methods mainly include differential centrifugation, density gradient centrifugation, SEC size exclusion chromatography and the like, wherein the differential centrifugation is the most common method, and generally, cell suspension is centrifuged at 3000rpm for 10min, supernatant solution (usually about 14000g) is taken and centrifuged for 2min, and the supernatant solution is centrifuged for 60min to obtain precipitate, namely the cell vesicles. However, the existing methods use equipment with high operation complexity and high cost, and the purification efficiency of the cell vesicles needs to be further improved.
SUMMERY OF THE UTILITY MODEL
The utility model provides a cell vesicle purification device, the device simple structure, easily operation, it is high to the extraction efficiency of cell vesicle.
An aspect of the utility model provides a cell vesicle purification device, include: the container comprises a container cover, a first container, a second container, a third container, a fourth container and a filtrate collecting part which are sequentially connected from top to bottom, wherein the first container is communicated with the second container through a first microporous filter membrane, the second container is communicated with the third container through a second microporous filter membrane, the third container is communicated with the fourth container through a third microporous filter membrane, the fourth container is communicated with the filtrate collecting part, the aperture of the first microporous filter membrane is 1-5 mu m, the aperture of the second microporous filter membrane is 500-1000nm, and the aperture of the third microporous filter membrane is 100-200 nm.
According to the utility model discloses an embodiment, install rabbling mechanism on the vessel cover, the rabbling mechanism extends to in the first container.
According to an embodiment of the utility model, be equipped with the filling opening that communicates with first container and the sealed lid that is used for the closing cap filling opening on the container lid.
According to an embodiment of the present invention, the ratio between the maximum radial dimension of the cross section of the first microporous membrane, the maximum radial dimension of the cross section of the second microporous membrane, and the maximum radial dimension of the cross section of the third microporous membrane is (8 ± 1): (5. + -. 1): 3.
according to the utility model discloses an embodiment, above-mentioned device still is equipped with the vacuum interface of bleeding with first container, second container, third container, fourth container intercommunication, and the position of vacuum interface of bleeding is less than third millipore filtration.
According to the utility model discloses an embodiment, the second container is including the first cavity portion that is located third millipore filtration membrane top and the second cavity portion that is located third millipore filtration membrane below, and the maximum radial dimension of cross section of first cavity portion is greater than the maximum radial dimension of cross section of second cavity portion, and third container, fourth container are located second cavity portion, and second cavity portion is equipped with the vacuum interface of bleeding.
According to the utility model discloses an embodiment, the lower extreme of fourth container is equipped with the liquid outlet, and the fourth container passes through liquid outlet and filtrating collecting part intercommunication, and the position of liquid outlet is less than the vacuum interface of bleeding.
According to an embodiment of the present invention, the liquid outlet is a bevel opening, and the opening of the bevel opening faces away from the vacuum pumping port.
According to the utility model discloses an embodiment, the fourth container includes the third cavity portion of being connected with the third container, is located the fourth cavity portion of third cavity portion below, is used for connecting the connecting portion of third cavity portion and fourth cavity portion, and the maximum radial dimension of the cross section of third cavity portion is greater than the maximum radial interface size of the cross section of fourth cavity portion, and the lower extreme of fourth cavity portion is equipped with the liquid outlet.
According to the utility model discloses an embodiment, can dismantle and sealing connection between container lid, first container, second container, third container, fourth container, the filtrating collection portion.
The utility model provides a cell vesicle purification device, based on the particle size of cell vesicle, utilize the millipore filtration of multiple different apertures to carry out level purification and handle, can effectively filter out the impurity of treating in the purification feed liquid, obtain the cell vesicle of high purity, and have draw advantages such as convenient and fast, device simple structure, with low costs.
Drawings
Fig. 1 is a schematic structural view of a cell vesicle purification apparatus according to an embodiment of the present invention;
fig. 2 is a schematic structural view of a filter unit according to an embodiment of the present invention;
fig. 3 is a schematic structural view of a container lid according to an embodiment of the present invention;
the reference numbers illustrate:
1. a first container; 2. a second container; 3. a third container; 4. a fourth container; 5. a filtrate collection section; 6. a container cover; 7. a vacuum air extraction interface; 11. a first microporous filtration membrane; 12. a second microporous filtration membrane; 13. a third microporous filtration membrane; 21. a first cavity portion; 22. a second chamber portion; 41. a third chamber portion; 42. a fourth cavity portion; 61. a stirring mechanism; 62. a liquid filling port; 421. and a liquid outlet.
Detailed Description
In order to make the technical solution of the present invention better understood, the present invention will be further described in detail with reference to the accompanying drawings.
In one aspect of the present invention, there is provided a cell vesicle purification apparatus, as shown in fig. 1 to 3, the apparatus comprising: the container comprises a container cover 6, a first container 1, a second container 2, a third container 3, a fourth container 4 and a filtrate collecting part 5 which are sequentially connected from top to bottom, wherein the first container 1 is communicated with the second container 2 through a first microporous filter membrane 11, the second container 2 is communicated with the third container 3 through a second microporous filter membrane 12, the third container 3 is communicated with the fourth container 4 through a third microporous filter membrane 13, the fourth container 4 is communicated with the filtrate collecting part 5, the aperture of the first microporous filter membrane 11 is 1-5 mu m, the aperture of the second microporous filter membrane 12 is 500-ion-exchange-1000 nm, and the aperture of the third microporous filter membrane 13 is 100-ion-exchange-200 nm.
The utility model discloses a cell vesicle purification device is to cell vesicle purification design, and its use is as follows: opening the container cover 6, injecting feed liquid to be purified containing cell vesicles into the first container 1, and filtering cell fragments with larger sizes from the feed liquid to be purified through the first microporous filter membrane 11 to obtain first filtrate; the first filtrate enters the second container 2 (for example, enters a second cavity part 21 of the second container 2), and then passes through a microporous filter membrane (ultrafiltration membrane) 12 to filter out cell debris with smaller size, so as to obtain a second filtrate; the second filtrate enters a third container 3, then passes through a third microporous filter membrane (ultrafiltration membrane) 13, so that cell vesicles (the minimum size is at least more than 150nm and most of the cell vesicles are more than 200 nm) are enriched on the third microporous filter membrane 13, and small-size impurities enter a fourth container 4 along with the obtained third filtrate and enter a filtrate collection part 3 after passing through the fourth container 4; and collecting the cell vesicles on the third microfiltration membrane 13, and flushing the third microfiltration membrane 13 with the trapped cell vesicles by using a flushing fluid (buffer solution) to obtain a cell vesicle suspension, so as to purify the cell vesicles.
The feed liquid to be purified containing cell vesicles can be obtained by a method that is conventional in the art, and for example, cell vesicles can be released from tumor cells by irradiating tumor cells with ultraviolet light or administering a chemotherapeutic drug to tumor cells as described in patent document CN102302784B, thereby obtaining a cell fluid containing cell vesicles (i.e., the feed liquid to be purified), but the feed liquid to be purified can also be obtained by other suitable methods. In specific implementation, the purification process may be performed by removing in advance impurities such as large cell debris in the cell fluid containing the cell vesicles in combination with centrifugation, and then using the supernatant obtained after centrifugation as the feed liquid to be purified.
The utility model discloses a cell vesicle purification device especially can carry out small-scale purification processing, for example be applied to the relevant research of cell vesicle in the laboratory, can reach high purification precision, satisfy the demand of experimental study to cell vesicle purity. Specifically, the throughput of the device of the present invention may be 1 × 10 total cells8-2*108Each time, namely, when one purification treatment is carried out, the total amount of the cell vesicles in the feed liquid to be purified, which is added into the device, is 1 x 10 times the total amount of the cells8-2*108The total amount of cell vesicles in the suspension obtained after the cell sap is treated by ultraviolet irradiation and the like.
In the present invention, the fixing/mounting manner of the first microporous filter membrane 11, the second microporous filter membrane 12 and the third microporous filter membrane 13 is not particularly limited, and conventional manners in the art can be adopted as long as the above-mentioned functions can be achieved, for example, in some embodiments, a first upper clamping plate is present at the bottom edge of the first container 1, a first lower clamping plate cooperating with the first upper clamping plate is present at the top edge of the second container 2, the edge of the first microporous filter membrane 11 is clamped in the interlayer formed by the first upper clamping plate and the first lower clamping plate, and the communicating portion between the first container 1 and the second container 2 is covered, so that the first container 1 and the second container 2 are communicated through the first microporous filter membrane 11 (specifically, the communicating portion is communicated through the rest portion except the edge portion where the first microporous filter membrane 11 is clamped by the interlayer); a second upper clamping plate is arranged at the bottom edge of the second container 2, a second lower clamping plate matched with the second upper clamping plate is arranged at the top edge of the third container 3, the edge of the second microporous filter membrane 12 is clamped in a sandwich layer formed by the second upper clamping plate and the second lower clamping plate, and a communication part between the second container 2 and the third container 3 is covered, so that the second container 2 and the third container 3 are communicated through the second microporous filter membrane 12 (specifically, the communication part is communicated through the rest part except the edge part of the second microporous filter membrane 12 clamped by the sandwich layer); the bottom edge of the third container 3 is provided with a third upper clamping plate, the top edge of the fourth container 4 is provided with a third lower clamping plate matched with the third upper clamping plate, the edge of the third microporous filter membrane 13 is clamped in a sandwich formed by the third upper clamping plate and the third lower clamping plate, and the communication part between the third container 3 and the fourth container 4 is covered, so that the third container 3 and the fourth container 4 are communicated through the third microporous filter membrane 13 (specifically, the communication part is communicated through the rest part except the edge part of the third microporous filter membrane 13 clamped by the sandwich).
In some embodiments, the pore size of the first microfiltration membrane 11 may be 1 μm, 2 μm, 3 μm, 4 μm, 5 μm or a range consisting of any two of these values; the pore size of the second microfiltration membrane 12 may be 500nm, 550nm, 600nm, 650nm, 700nm, 750nm, 800nm, 850nm, 900nm, 950nm, 1000nm or a range consisting of any two of these values; the aperture of the third microfiltration membrane 13 can be 100nm, 120nm, 140nm, 160nm, 180nm, 200nm or a range consisting of any two of the numerical values, which is beneficial to improving the purification efficiency.
In some embodiments, the container cover 6 is provided with a stirring mechanism 61, the stirring mechanism 61 extends into the first container 1, and when the purification treatment is performed, the stirring mechanism 61 can be opened to stir the feed liquid in the first container 1, so as to prevent the larger cell debris in the feed liquid from settling and accumulating to block the pores of the first microporous filtering membrane 11, thereby further improving the purification efficiency. Specifically, the upper end of the stirring structure 61 is mounted on the container cover 6, and the lower end thereof is provided with a stirring member (such as a propeller) positioned in the first container 1 to stir the feed liquid in the first container 1. The stirring structure may be a stirrer conventional in the art as long as the above-described stirring function can be achieved.
In some embodiments, the container cover 6 is provided with a filling opening 62 communicated with the first container 1 and a sealing cover for sealing the filling opening 62, a washing liquid (such as a buffer solution) can be added into the first container 1 through the filling opening 62 to facilitate washing of filter residues on the first microporous filter membrane 11, so that more cell vesicles can pass through the first microporous filter membrane 11, the purification efficiency is improved, of course, other required liquid can also be added through the filling opening 62 as required, for example, to-be-purified liquid is added or supplemented into the first container 1, and after the charging is completed, the filling opening 62 can be sealed through the sealing cover to facilitate the purification process.
In some embodiments, the maximum radial dimensions of the cross sections of the first microporous filter membrane 11, the second microporous filter membrane 12 and the third microporous filter membrane 13 are sequentially reduced, which is beneficial to the hierarchical separation and purification of the feed liquid to be purified, so that the cell vesicles are finally enriched/concentrated on the smaller third microporous filter membrane 13, and the separated cell vesicles are convenient to collect.
Specifically, in some preferred embodiments, the ratio between the cross-sectional maximum radial dimension of the first microfiltration membrane 11, the cross-sectional maximum radial dimension of the second microfiltration membrane 12 and the cross-sectional maximum radial dimension of the third microfiltration membrane 13 is (8 ± 1): (5. + -. 1): 3, for example, the maximum radial dimension of the cross section of the first microfiltration membrane 11 is 8. + -.1 cm, the maximum radial dimension of the cross section of the second microfiltration membrane 12 is 5. + -.1 cm, and the maximum radial dimension of the cross section of the third microfiltration membrane 13 is 3 cm.
Typically, the first microfiltration membrane 11, the second microfiltration membrane 12 and the third microfiltration membrane 13 are arranged coaxially and have substantially the same cross-sectional shape, for example, a circular shape, the largest radial dimension of the cross-section being the diameter of the cross-section.
In some embodiments, the utility model discloses a still be equipped with on the device with first container 1, second container 2, third container 3, the vacuum interface of bleeding (or called the mouth of bleeding) 7 of fourth container 4 intercommunication, the position of vacuum interface of bleeding 7 is less than third microfiltration membrane 13 (vacuum interface of bleeding 7 is located the below of third microfiltration membrane 13 promptly), vacuum interface of bleeding 7 is used for linking to each other with the pump system (or called the vacuum pump) that has the evacuation function, provide the negative pressure through the pump system, to carrying out evacuation processing in the container, do benefit to the purification in-process and wait to purify the feed liquid and pass through first microfiltration membrane 11 in proper order, second microfiltration membrane 12 and second microfiltration membrane 13, further improve purification efficiency. The pump system may be manual or electric, and the present invention is not limited thereto.
Specifically, in some embodiments, the second container 2 includes a first cavity portion 21 located above the third microfiltration membrane 13 and a second cavity portion 22 located below the third microfiltration membrane 13, the maximum radial dimension of the cross section of the first cavity portion 21 is larger than the maximum radial dimension of the cross section of the second cavity portion 22, the third container 3 and the fourth container 4 are located in the second cavity portion 22, and the second cavity portion 22 is provided with the vacuum pumping interface 7, so that the purification efficiency can be further improved, and the device structure can be simplified. The first cavity portion 21 and the second cavity portion 21 have substantially the same cross-sectional shape, for example, circular.
In some embodiments, the volume of the first container 1 is greater than the volume of the first cavity portion 21 of the second container 2, and the volume of the second cavity portion 21 is greater than the volume of the third container 3, which may be set as desired, for example, the volume of the first container may be 250 ± 50 mL.
In some embodiments, a liquid outlet 421 is disposed at a lower end of the fourth container 4, the fourth container 4 is communicated with the filtrate collecting portion 5 through the liquid outlet 421, and the position of the liquid outlet 421 is lower than the vacuum pumping port 7, so that the filtrate in the container can be prevented from being pumped out during the vacuum pumping process, and the purification process can be performed more conveniently.
Further, the liquid outlet 421 is an oblique opening, and an opening of the oblique opening faces away from the vacuum pumping port 7 (i.e. the liquid outlet 421 faces to the other side opposite to the side where the vacuum pumping port 7 is located), so that the filtrate in the container can be further prevented from being pumped out during the vacuum pumping process. Specifically, the inclination of the liquid outlet 421 may be 30 ° to 60 ° (i.e., the angle between the cross-section of the liquid outlet 421 and the horizontal direction is 30 ° to 60 °), for example, 45 °.
Specifically, in some embodiments, the fourth container 4 includes a third cavity portion 41 connected to the third container 3, a fourth cavity portion 42 located below the third cavity portion 41, and a connecting portion for connecting the third cavity portion 41 and the fourth cavity portion 42, a maximum radial dimension of a cross section of the third cavity portion 41 is larger than a maximum radial dimension of a cross section of the fourth cavity portion 42, and a lower end of the fourth cavity portion 42 is provided with the liquid outlet 421. The third cavity portion 41, the fourth cavity portion 42 and the connecting portion are coaxially arranged, and the cross-sectional shapes thereof are substantially the same, for example, circular. As shown in fig. 1 and 2, in some embodiments, the fourth container 4 is funnel-shaped.
In some embodiments, the container cover 6, the first container 1, the second container 2, the third container 3, the fourth container 4, and the filtrate collecting part 5 are detachably and hermetically connected, and these components may be coaxially disposed, and the cross-sectional shapes of the mutual connection portions are substantially the same, such as circular. For example, in some embodiments, the bottom end of the third container 3 is connected to the upper end of the fourth container 4, the maximum radial dimension of the cross section of the third container 3 decreases from top to bottom, the maximum radial dimension of the cross section of the bottom end is the same as the maximum radial dimension of the cross section of the upper end of the fourth container, and the cross section has substantially the same shape, such as a circle.
In some embodiments, the container cover 6 may be snapped onto the first container 1, the second container 2, the third container 3, and the fourth container 4 may be connected by threads, and the fourth container 4 and the filtrate collecting portion 5 may be connected by a frosted opening, specifically, the upper portion of the filtrate collecting portion 5 has a frosted opening, the second container 2 has the first cavity portion 21 and the second cavity portion 22, the second cavity portion 22 has a frosted portion matching with the frosted opening, and the frosted portion is inserted into the frosted opening to realize detachable connection of the fourth container 4 and the filtrate collecting portion 5, and through the above connection manner, sealed connection of the components is realized, and a cell vesicle purification device is formed.
In some embodiments, the first container 1, the second container 2, the third container 3, the fourth container 4, and the filtrate collecting unit 5 may be made of glass and/or plastic, and in particular, transparent materials may be selected as needed to facilitate observation of the purification process of the cell vesicles.
In the present invention, if there is no specific description, the components such as the first container 1, the second container 2, the third container 3, the fourth container 4, the filtrate collecting portion 5 can be conventional instruments/components/structures used in the field as long as the corresponding functions can be realized, for example, the filtrate collecting portion can be a conventional filtrate collecting bottle such as a conical bottle, and the device of the present invention can be assembled by these components according to the conventional method in the field.
To make the objects, technical solutions and advantages of the present invention clearer, embodiments of the present invention are combined to clearly and completely describe the technical solutions in the embodiments of the present invention, and obviously, the described embodiments are some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
Example 1
The apparatus used in this embodiment is shown in fig. 1, and includes a container body and a container lid 1 for closing the container body, the container body including: the device comprises a container cover 6, a first container 1, a second container 2, a third container 3, a fourth container 4 and a filtrate collecting part 5 which are sequentially connected from top to bottom, wherein the first container 1 is communicated with the second container 2 through a first microporous filter membrane 11, the second container 2 is communicated with the third container 3 through a second microporous filter membrane 12, the third container 3 is communicated with the fourth container 4 through a third microporous filter membrane 13, the fourth container 4 is communicated with the filtrate collecting part 5, the pore diameter of the first microporous filter membrane 11 is 3 mu m, the pore diameter of the second microporous filter membrane 12 is 500nm, and the pore diameter of the third microporous filter membrane 13 is 150 nm; the first microporous filter membrane 11, the second microporous filter membrane 12 and the third microporous filter membrane 13 are coaxially arranged, the cross section shapes of the first microporous filter membrane 11, the second microporous filter membrane 12 and the third microporous filter membrane 13 are all circular, the cross section diameter of the first microporous filter membrane 11 is 8cm, the cross section diameter of the second microporous filter membrane 12 is 5cm, and the cross section diameter of the third microporous filter membrane 13 is 3 cm;
the container cover 6 is provided with a stirring mechanism 61, and the stirring mechanism 61 extends into the first container 1 and is used for stirring the feed liquid in the first container 1; the container cover 6 is provided with a filling port 62 communicated with the first container 1 and a sealing cover for sealing the filling port 62, and the filling port 62 is used for adding flushing liquid (such as buffer liquid) into the first container 1;
the second container 2 comprises a first cavity part 21 positioned above the third microporous filter membrane 13 and a second cavity part 22 positioned below the third microporous filter membrane 13, the cross sections of the first cavity part 21 and the second cavity part 22 are both circular, the diameter of the cross section of the first cavity part 21 is larger than that of the cross section of the second cavity part 22, the third container 3 and the fourth container 4 are positioned in the second cavity part 22, and the second cavity part 22 is provided with a vacuum pumping interface 7; the volume of the first container is 250mL, the volume of the first container 1 is larger than that of the first cavity 21 of the second container 2, and the volume of the second cavity 21 is larger than that of the third container 3;
the fourth container 4 is funnel-shaped, and comprises a third cavity part 41 connected with the third container 3, a fourth cavity part 42 positioned below the third cavity part 41, and a connecting part for connecting the third cavity part 41 and the fourth cavity part 42, wherein the third cavity part 41, the fourth cavity part 42 and the connecting part are coaxially arranged, the cross section of the third cavity part 41 is circular, the cross section diameter of the third cavity part 41 is larger than that of the fourth cavity part 42, a liquid outlet 421 is arranged at the lower end of the fourth cavity part 42, the fourth container 4 is communicated with the filtrate collecting part 5 through the liquid outlet 421, the position of the liquid outlet 421 is lower than the vacuum air suction interface 7, the liquid outlet 421 is an oblique opening, the inclination is 45 degrees, and the opening faces away from the vacuum air suction interface 7;
the container cover 6, the first container 1, the second container 2, the third container 3, the fourth container 4 and the filtrate collecting part 5 are detachably and hermetically connected, the components are coaxially arranged, and the cross section of the connected parts is circular.
Application example 1
1 x 10 of8Suspending A549 cells in 20mL 1640 culture medium, mixing, treating with ultraviolet radiation for 60min, and placing in CO2Incubating in an incubator for 16 hours to obtain a cell suspension; placing the cell suspension in a 50mL centrifuge tube, rinsing the culture dish with 10mL Phosphate Buffered Saline (PBS), combining the obtained washing solution into the centrifuge tube, centrifuging at 3000rpm for 10min to obtain a supernatant, taking the supernatant as a feed liquid to be purified, and purifying the cell vesicles by adopting the device in example 1;
adding the supernatant into a first container 1, adding 100mL of PBS into the first container, covering a container cover 6, opening a stirring mechanism 61, opening a vacuum pump communicated with a vacuum pumping interface 7 to start pumping filtration, and adding 200mL of PBS from a liquid adding port 62 in batches during the pumping filtration; when the liquid level of the feed liquid in the first container 1 is reduced to be below a propeller of the stirring mechanism 61, closing the stirring mechanism 61;
after all the supernatant is filtered to the filtrate collecting part 5, the vacuum pump is closed, and the filtration is finished; disassembling the device, taking the third microporous filter membrane 13, taking about 1mL of PBS by using a pipette, repeatedly washing the surface of the third microporous filter membrane adsorbing the cell vesicles, and collecting eluent to obtain the cell vesicle suspension.
The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A cell vesicle purification apparatus, comprising: the container comprises a container cover, a first container, a second container, a third container, a fourth container and a filtrate collecting part which are sequentially connected from top to bottom, wherein the first container is communicated with the second container through a first microporous filter membrane, the second container is communicated with the third container through a second microporous filter membrane, the third container is communicated with the fourth container through a third microporous filter membrane, the fourth container is communicated with the filtrate collecting part, the aperture of the first microporous filter membrane is 1-5 mu m, the aperture of the second microporous filter membrane is 500-1000nm, and the aperture of the third microporous filter membrane is 100-200 nm.
2. The apparatus for purifying cell vesicles according to claim 1, wherein a stirring mechanism is attached to the container lid, and the stirring mechanism extends into the first container.
3. The apparatus for purifying cell vesicles according to claim 1, wherein the container cover is provided with a filling port communicating with the first container and a sealing cover for sealing the filling port.
4. The cellular vesicle purification apparatus according to claim 1, wherein the ratio of the maximum radial dimension of the cross section of the first microfiltration membrane to the maximum radial dimension of the cross section of the second microfiltration membrane to the maximum radial dimension of the cross section of the third microfiltration membrane is (8 ± 1): (5. + -. 1): 3.
5. the apparatus for purifying cell vesicles according to claim 1, further comprising a vacuum pumping port communicating with the first container, the second container, the third container, and the fourth container, wherein the vacuum pumping port is located lower than a third microporous filter membrane.
6. The apparatus of claim 5, wherein the second container comprises a first chamber portion located above the third microfiltration membrane and a second chamber portion located below the third microfiltration membrane, the first chamber portion having a larger maximum radial dimension in cross section than the second chamber portion, the third and fourth containers being located in the second chamber portion, and the second chamber portion being provided with the vacuum pumping port.
7. The cell vesicle purification apparatus according to claim 4, wherein a liquid outlet is provided at a lower end of the fourth container, the fourth container is communicated with the filtrate collection portion through the liquid outlet, and the liquid outlet is positioned lower than the vacuum pumping port.
8. The apparatus of claim 7, wherein the liquid outlet is a beveled opening with an opening facing away from the vacuum pumping port.
9. The cellular vesicle purification apparatus according to claim 7, wherein the fourth container comprises a third cavity portion connected to the third container, a fourth cavity portion located below the third cavity portion, and a connecting portion for connecting the third cavity portion and the fourth cavity portion, wherein the third cavity portion has a maximum radial cross-sectional dimension larger than a maximum radial cross-sectional dimension of the fourth cavity portion, and the lower end of the fourth cavity portion is provided with the liquid outlet.
10. The apparatus for purifying cell vesicles according to any one of claims 1 to 9, wherein the container cover, the first container, the second container, the third container, the fourth container, and the filtrate collection unit are detachably and hermetically connected.
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| CN202120316035.1U CN214552580U (en) | 2021-02-04 | 2021-02-04 | Cell vesicle purification device |
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| CN202120316035.1U CN214552580U (en) | 2021-02-04 | 2021-02-04 | Cell vesicle purification device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2024023501A3 (en) * | 2022-07-25 | 2024-02-29 | Micropore Technologies Ltd. | Continuous process |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2024023501A3 (en) * | 2022-07-25 | 2024-02-29 | Micropore Technologies Ltd. | Continuous process |
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