CN210700184U - Exosome filters extraction element - Google Patents
Exosome filters extraction element Download PDFInfo
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- CN210700184U CN210700184U CN201921178794.5U CN201921178794U CN210700184U CN 210700184 U CN210700184 U CN 210700184U CN 201921178794 U CN201921178794 U CN 201921178794U CN 210700184 U CN210700184 U CN 210700184U
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Abstract
The utility model provides an exosome filtering and extracting device, which comprises an outer centrifuge tube, an inner centrifuge tube, a first filter membrane and a second filter membrane; the inner centrifugal tube is arranged in the inner cavity of the outer centrifugal tube; an injection pipe is inserted into the upper end surface of the inner centrifugal pipe and penetrates through the upper end surface of the outer centrifugal pipe, so that the inner cavity of the inner centrifugal pipe is communicated with the outside of the device; the lower end surface of the inner centrifugal tube is provided with a discharge port; the first filter membranes with different pore sizes are arranged in the inner cavity of the inner centrifuge tube at equal intervals, and the first filter membranes are sequentially arranged in the inner cavity of the inner centrifuge tube in the direction from the injection tube to the discharge port according to the order of the pore sizes from large to small; the second filter membrane is erected between the outer side wall of the inner centrifugal tube and the inner side wall of the outer centrifugal tube and arranged around the outer side wall of the inner centrifugal tube for a circle; the bottom of outer centrifuging tube is equipped with gets the material mouth to provide one kind and guarantee aseptic environment and make exosome filter and draw simple exosome of easy operation and filter extraction device and exosome extraction method.
Description
Technical Field
The utility model relates to an exosome draws technical field, especially relates to an exosome filters extraction element.
Background
Extracellular Vesicles (EVs) are a highly conserved intercellular communication system through which cells exchange information in the form of lipids, proteins or nucleic acids. EVs were initially found to be involved in bone mineralization as well as platelet function, known as "platelet dust". Given their extremely broad biological function and their ability to shuttle macromolecules between cells, EVs provide a unique platform for developing a new class of therapeutics.
Exosomes (Exos) are a subset of EVs, present in all body fluids, and are vesicles (30nm-150nm) released from all intracellular parts of the human body via the multivesicular endosomal pathway. Exosomes are the most promising type of EVs for therapeutic prospects.
Sources of exosomes are commonly cultured cells in vitro and human-derived body fluids. The biggest problem with exosomes derived from cells is that the use of cells from others is ethical; the culture period of the cells using the cells is too long, and the uncertainty factor is too large. The exosome derived from human body fluid can well avoid the problems, and the exosome derived from the body fluid of the patient is separated, processed and extracted and then used for the patient, so that the problem in the aspect of ethics does not exist.
The most common of the body fluids is blood. The serum exosome not only contains exosome in blood, but also contains exosome from platelet source, and has good clinical prospect. Various exosomes of humoral origin, including serum exosomes and exosomes from platelet rich plasma, have been shown in considerable literature to promote tissue repair and tissue protection functions. The early researches also find that the exosomes from the body fluid sources have good functions of promoting tissue repair and preventing tissue and organ damage caused by various reasons, and have good clinical application prospects.
However, the traditional method for extracting exosome has disadvantages in clinical use. Most exosome extraction methods are now used mainly in the laboratory. Large instruments are required, a sterile operating room is required, and operation is performed by specially trained workers. If the exosome is put into clinical use, the flow needs to be simplified, and the admission threshold is reduced, so that the exosome cannot be extracted by matching with so many specially trained talents for clinical use.
In general, the products we need to be clinically useful must have two features: easy to handle and maintain sterile.
In laboratory studies, ultrafiltration is well-established and relatively simple to operate. However, even with ultrafiltration, sterility is achieved in the laboratory by professionals in the clean bench using pipettes, sterile tips and various sterile consumables, which not only impose requirements on hardware conditions, but also on the skills of the relevant operators. In order to be popularized clinically, devices such as pipettors are avoided as much as possible, operation is reduced, most of operation is completed in one tube, sterility in clinical application can be guaranteed, and learning cost can be reduced. The skill of the relevant operator for obtaining the exosome for treatment aseptically can be mastered only by short-term training.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide a guarantee sterile environment and make exosome filter and draw simple easy operation's exosome filtration extraction element.
In order to achieve the above purpose, the utility model provides an exosome filtration and extraction device, which comprises an outer centrifuge tube, an inner centrifuge tube, a first filter membrane and a second filter membrane; the inner centrifugal tube is arranged in the inner cavity of the outer centrifugal tube; an injection pipe is inserted into the upper end surface of the inner centrifugal pipe and penetrates through the upper end surface of the outer centrifugal pipe, so that the inner cavity of the inner centrifugal pipe is communicated with the outside of the device; the lower end surface of the inner centrifugal tube is provided with a discharge port;
the first filter membranes with different pore sizes are arranged in the inner cavity of the inner centrifuge tube at equal intervals, and the first filter membranes are sequentially arranged in the inner cavity of the inner centrifuge tube in the direction from the injection tube to the discharge port in the order from the large pore size to the small pore size;
the second filter membrane is erected between the outer side wall of the inner centrifugal tube and the inner side wall of the outer centrifugal tube, and is arranged around the outer side wall of the inner centrifugal tube for a circle; and a material taking port is arranged at the bottom end of the outer centrifuge tube.
Preferably, the first filter membrane is four, and the pore diameters are respectively 10 μm, 5 μm, 1 μm and 200 nm.
Preferably, the periphery of first filter membrane with a week of the inside wall of interior centrifuging tube is contradicted each other, avoids first filter membrane with appear the clearance between the inside wall of interior centrifuging tube.
Preferably, the vertical section of the second filter membrane is in a groove shape; the groove of the second filter membrane is arranged facing the material taking port; the side of slot both sides respectively with the lateral wall of interior centrifuging tube with the inside wall of outer centrifuging tube is contradicted each other, avoids the second filter membrane with the lateral wall of interior centrifuging tube with appear the clearance between the inside wall of outer centrifuging tube.
Preferably, the device further comprises a capillary tube, wherein the capillary tube extends into the inner cavity of the outer centrifugal tube through the material taking port to suck the exosomes on the second filter membrane out.
Preferably, the device also comprises a conical base; the bottom surface of the base is connected with the upper end face and the lower end face of the outer centrifugal tube in a matched mode.
Preferably, the injection pipe and the material taking port are both provided with detachable sealing plugs.
Preferably, the outer centrifuge tube and the inner centrifuge tube are both cylinders.
Compared with the prior art, the utility model has the advantages that: the utility model discloses an exosome filters extraction element makes the body fluid carry out twice filtration, thereby once only can obtain the exosome of appointed specification in exosome filters extraction element, the adoption multiple apparatus filters operation steps such as drawing in having abandoned traditional ultrafiltration method, aseptic environment has been guaranteed, the exosome filters the quality of drawing has been improved, the reduction of concurrent operation step and use equipment, the operation degree of difficulty that the exosome drawed has also been reduced, the simple easy operation of whole set of flow, easily grasp, do not need the professional to go to operate, the degree of difficulty of personnel's configuration has been reduced, be convenient for at clinical use and popularization.
Drawings
Fig. 1 is a schematic structural diagram of an exosome filtration-extraction device in an embodiment of the present invention;
fig. 2 is a schematic structural view of the lower end surface of the exosome filtering and extracting device combined with a conical base in an embodiment of the present invention;
fig. 3 is a schematic structural view of the upper end surface of the exosome filtering and extracting device combined with the conical base in an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be further described below.
As shown in fig. 1, the present invention aims to provide an exosome filtering and extracting device and an exosome extracting method which ensure sterile environment and make exosome filtering and extracting simple and easy to operate.
In order to achieve the purpose, the utility model provides an exosome filtration and extraction device, which comprises an outer centrifuge tube 1, an inner centrifuge tube 2, a first filter membrane 5 and a second filter membrane 6; the inner centrifuge tube 2 is arranged in the inner cavity of the outer centrifuge tube 1; an injection pipe 3 is inserted into the upper end face of the inner centrifugal tube 2, and the injection pipe 3 penetrates through the upper end face of the outer centrifugal tube 1, so that the inner cavity of the inner centrifugal tube 2 is communicated with the outside of the device; the lower end surface of the inner centrifuge tube 2 is provided with a discharge port;
the first filter membranes 5 with different pore sizes are arranged in the inner cavity of the inner centrifuge tube 2 at equal intervals, and the first filter membranes 5 are sequentially arranged in the inner cavity of the inner centrifuge tube 2 from the injection tube 3 to the discharge port in the order of the pore sizes from large to small;
the second filter membrane 6 is erected between the outer side wall of the inner centrifugal tube 2 and the inner side wall of the outer centrifugal tube 1, and the second filter membrane 6 is arranged around the outer side wall of the inner centrifugal tube 2 for a circle; the bottom of outer centrifuging tube 1 is equipped with gets material mouth 4.
In this embodiment, the first filter 5 has four filter pores with a diameter of 10 μm, 5 μm, 1 μm and 200 nm.
In this embodiment, the periphery of first filter membrane 5 contradicts each other with a week of interior centrifuging tube 2's inside wall, avoids appearing the clearance between the inside wall of first filter membrane 5 and interior centrifuging tube 2, leads to first filter membrane 5's filter effect not good for major diameter granule gets into 1 inner chamber of outer centrifuging tube in the body fluid.
In this embodiment, the vertical section of the second filter membrane 6 is groove-shaped; the groove of the second filter membrane 6 faces the material taking port 4; the side edges of the two sides of the groove are respectively butted with the outer side wall of the inner centrifugal tube 2 and the inner side wall of the outer centrifugal tube 1, so that a gap is prevented from being formed between the second filter membrane 6 and the outer side wall of the inner centrifugal tube 2 and the inner side wall of the outer centrifugal tube 1, exosomes in a specified specification are discharged from the system, and the harvesting efficiency is reduced; the design of the groove shape enables exosomes within the specified specification to be collected and concentrated in the groove, and further collection of the capillary 8 is facilitated.
In this embodiment, the device further comprises a capillary tube 8, wherein the capillary tube 8 extends into the inner cavity of the outer centrifugal tube 1 through the material taking port 4 to suck the exosome on the second filter membrane 6 out.
In the present embodiment, as shown in fig. 2 and 3, the device further includes a conical base 7; the bottom surface of the base 7 is matched and connected with the upper end surface and the lower end surface of the outer centrifuge tube 1, the side walls of the upper end surface and the lower end surface can be correspondingly provided with a plurality of external threads, the inner surface of the base 7 can be correspondingly provided with a plurality of internal threads, and the base 7 is matched and connected with the outer centrifuge tube 1 in a detachable way through the matching of the internal threads and the external threads; can dismantle coniform base 7 and be convenient for to fix the base cooperation in outer centrifuging tube 1 and the centrifuge to in being located the centrifuge with outer centrifuging tube 1, at centrifugal in-process, prevent that outer centrifuging tube 1 from taking place the dislocation in the centrifuge.
In this embodiment, the filling tube 3 and the extraction opening 4 are each provided with a removable sealing plug.
In this embodiment, the outer centrifuge tube 1 and the inner centrifuge tube 2 are both cylindrical.
An exosome extraction method, comprising the following steps:
in order to facilitate understanding of the use of the device, the utility model also provides an exosome extraction method, which comprises the following steps:
step 1: taking a sealed exosome filtering and extracting device; ensuring that the exosome filtering and extracting device is in an aseptic environment at first;
step 2: taking out the sealing plug on the injection tube 3, and injecting the body fluid into the inner cavity of the inner centrifugal tube 2 through the injection tube 3;
and step 3: the sealing plug on the injection pipe 3 is covered;
and 4, step 4: the bottom surface of the conical base is matched and connected with the lower end surface of the outer centrifuge tube 1, so that the outer centrifuge tube 1 is placed in a centrifuge for centrifugation; after the centrifugal machine is opened, the body fluid is filtered by a first filter membrane 5 with the thickness of 10 microns, 5 microns, 1 micron and 200nm in sequence; large particles and large impurities in the body fluid are removed, so that the body fluid with the diameter less than 200nm enters the inner cavity of the outer centrifugal tube 1, as shown in fig. 2.
And 5: centrifuging, taking out the exosome filtration and extraction device, and inverting the exosome filtration and extraction device;
step 6: taking down the base from the lower end face of the outer centrifuge tube 1, and connecting the base with the upper end face of the outer centrifuge tube 1 in a matching manner, so that the outer centrifuge tube 1 is placed upside down in a centrifuge for centrifugation; after the centrifuge is opened, the exosomes with the diameter less than 200nm and more than 30nm are filtered on the surface of the second filter membrane 6, and then the water and the proteins with smaller diameters are filtered, as shown in fig. 3.
And 7: after centrifugation, taking out the exosome filtration and extraction device, taking down the base from the outer centrifuge tube 1, and opening the sealing plug of the discharge port;
and 8: get capillary 8 and stretch into outer centrifuging tube 1 inner chamber from the discharge port, take out the exosome on the second filter membrane 6, guarantee that whole centrifugal filtration and extraction step all accomplish in a device, liquid keeps isolation state with the external world basically all the time to fully guarantee the aseptic of device and whole operation process.
In step 7, the exosome filtration extraction device is removed after centrifugation and the base is removed from the outer centrifuge tube 1, opening the vent closure.
In this embodiment, the utility model discloses an exosome filters extraction element makes the body fluid carry out twice filtration to once only can obtain the exosome of appointed specification in exosome filters extraction element, abandoned and adopted multiple apparatus to filter operation steps such as drawing in traditional ultrafiltration method, the utility model discloses whole operation is accomplished in a device, collects multiple filtration and integrative setting, has guaranteed aseptic environment, has improved exosome and has filtered the quality of drawing, and the reduction of concurrent operation step and use equipment has also reduced the operation degree of difficulty that exosome drawed, and whole set of flow is simple easy to operate, grasps easily, does not need the professional to go to operate, is convenient for at clinical use and popularization.
The above description is only for the preferred embodiment of the present invention, and does not limit the present invention. Any technical personnel who belongs to the technical field, in the scope that does not deviate from the technical scheme of the utility model, to the technical scheme and the technical content that the utility model discloses expose do the change such as the equivalent replacement of any form or modification, all belong to the content that does not break away from the technical scheme of the utility model, still belong to within the scope of protection of the utility model.
Claims (8)
1. An exosome filtration and extraction device is characterized by comprising an outer centrifuge tube, an inner centrifuge tube, a first filter membrane and a second filter membrane; the inner centrifugal tube is arranged in the inner cavity of the outer centrifugal tube; an injection pipe is inserted into the upper end surface of the inner centrifugal pipe and penetrates through the upper end surface of the outer centrifugal pipe, so that the inner cavity of the inner centrifugal pipe is communicated with the outside of the device; the lower end surface of the inner centrifugal tube is provided with a discharge port;
the first filter membranes with different pore sizes are arranged in the inner cavity of the inner centrifuge tube at equal intervals, and the first filter membranes are sequentially arranged in the inner cavity of the inner centrifuge tube in the direction from the injection tube to the discharge port in the order from the large pore size to the small pore size;
the second filter membrane is erected between the outer side wall of the inner centrifugal tube and the inner side wall of the outer centrifugal tube, and is arranged around the outer side wall of the inner centrifugal tube for a circle; and a material taking port is arranged at the bottom end of the outer centrifuge tube.
2. The exosome filtration-extraction device according to claim 1, characterized in that said first filter membrane is four with pore sizes of 10 μ ι η, 5 μ ι η, 1 μ ι η and 200nm, respectively.
3. The exosome filtration and extraction device according to claim 1, wherein the outer perimeter of the first filter membrane interferes with a circle of the inner side wall of the inner centrifuge tube, avoiding a gap between the first filter membrane and the inner side wall of the inner centrifuge tube.
4. The exosome filtration-extraction device according to claim 1, wherein the vertical cross-section of the second filter membrane is gutter-shaped; the groove of the second filter membrane is arranged facing the material taking port; the side of slot both sides respectively with the lateral wall of interior centrifuging tube with the inside wall of outer centrifuging tube is contradicted each other, avoids the second filter membrane with the lateral wall of interior centrifuging tube with appear the clearance between the inside wall of outer centrifuging tube.
5. The exosome filtration-extraction device according to claim 1, further comprising a capillary tube extending into the inner cavity of the outer centrifugal tube through the extraction port to suck out exosomes on the second filter membrane.
6. The exosome-filtering extraction device according to claim 1, further comprising a conical base; the bottom surface of the base is connected with the upper end face and the lower end face of the outer centrifugal tube in a matched mode.
7. The exosome filtration-extraction device according to claim 1, characterized in that the injection tube and the take-off port are each provided with a removable sealing plug.
8. The exosome-filtration extraction device according to claim 1, wherein the outer centrifuge tube and the inner centrifuge tube are both cylindrical.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921178794.5U CN210700184U (en) | 2019-07-25 | 2019-07-25 | Exosome filters extraction element |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921178794.5U CN210700184U (en) | 2019-07-25 | 2019-07-25 | Exosome filters extraction element |
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| CN210700184U true CN210700184U (en) | 2020-06-09 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110252440A (en) * | 2019-07-25 | 2019-09-20 | 上海市第六人民医院 | A kind of excretion body filtering extraction element and excretion body extracting method |
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2019
- 2019-07-25 CN CN201921178794.5U patent/CN210700184U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110252440A (en) * | 2019-07-25 | 2019-09-20 | 上海市第六人民医院 | A kind of excretion body filtering extraction element and excretion body extracting method |
| CN110252440B (en) * | 2019-07-25 | 2024-01-23 | 上海市第六人民医院 | Exosome filtering and extracting device and exosome extracting method |
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