Exosome concentrating, purifying and filtering device

Abstract

The utility model discloses an exosome concentrating, purifying and filtering device which consists of a first tank body, a second tank body, a sterile bottle, a feeding pipe, a first filtering bag, a second filtering bag, a first conveying pipeline and a second conveying pipeline, wherein a first filter bag is arranged in the first tank body, and a second filter bag is arranged in the second tank body; the first filter bag is communicated with the second filter bag through a first conveying pipeline, and the second filter bag is communicated with the sterile bottle through a second conveying pipeline. According to the exosome concentrating, purifying and filtering device disclosed by the utility model, the exosome is separated and purified through the multi-stage filter membrane and is used for separating and purifying the exosome, and the first filter bag or the second filter bag of the device is separated from the first tank body or the second tank body or can be detached from the first tank body or the second tank body to be replaced by a new filter bag; the cost can be obviously reduced, and the efficiency of obtaining and separating the exosome is improved.

Description

Technical Field

[0001] This utility model relates to a device for separating and purifying cell secretions, and more particularly to a device for concentrating, purifying and filtering exosomes, belonging to the field of exosome concentration, purification and filtration devices. Background Technology

[0002] Exosomes are nanoscale vesicles secreted by cells, typically ranging from 30 to 150 nanometers in diameter. They are widely distributed and found in human urine, sweat, blood, and other fluids. Exosomes consist of a lipid bilayer and contain cell surface proteins, intracellular proteins, RNA, DNA, amino acids, and metabolites. Exosomes have two main functions: first, information transmission, transferring molecules such as proteins, RNA, and DNA from one cell to another, thereby regulating the function of the recipient cell; second, genetic material transfer, delivering large amounts of RNA from one cell to another.

[0003] With the continuous deepening of exosome research and technological advancements, its application prospects are becoming increasingly broad. Early diagnosis and monitoring of diseases can be achieved by detecting exosomes and the molecules they carry in bodily fluids. The ability of exosomes to promote cell regeneration and tissue repair allows for their application in regenerative therapy. Their small size, strong penetrability, and high stability make them suitable for drug delivery. However, due to their small size and complex composition, efficient and high-purity extraction of exosomes has always been a challenge for both research and clinical applications.

[0004] Existing methods for extracting exosomes often suffer from problems such as complex operation, high cost, low extraction efficiency, and low purity, and urgently need improvement. Utility Model Content

[0005] The purpose of this invention is to provide an efficient, convenient, and low-cost exosome purification, concentration, filtration, and extraction device to solve the problems of high cost and low efficiency in the exosome extraction process.

[0006] To achieve the above objectives, the main technical solutions adopted by this utility model include:

[0007] An exosome concentration, purification, and filtration device comprises a first tank, a second tank, a sterile bottle, an inlet pipe, a first filter bag, a second filter bag, a first conveying pipe, and a second conveying pipe; wherein, the first filter bag is housed in the first tank, and the second filter bag is housed in the second tank; the first filter bag and the second filter bag are connected through the first conveying pipe, and the second filter bag and the sterile bottle are connected through the second conveying pipe.

[0008] In a preferred embodiment of this utility model, a stirrer is provided inside the first tank.

[0009] In a preferred embodiment of this utility model, a stirrer is provided at the bottom of the first tank.

[0010] In a preferred embodiment of this utility model, a feed pipe is provided on the first filter bag, and a feed pipe switch is provided on the feed pipe.

[0011] In a preferred embodiment of this utility model, a first peristaltic pump is provided on the first material conveying pipe.

[0012] In a preferred embodiment of this utility model, a second peristaltic pump is provided on the second material conveying pipeline.

[0013] In a preferred embodiment of this utility model, the first filter bag or the second filter bag is separable from the first tank or the second tank or can be disassembled and removed from the first tank or the second tank. Therefore, the used first filter bag or the second filter bag can be removed from the first tank or the second tank and replaced with a new first filter bag or the second filter bag.

[0014] For reference, the method for achieving exosome concentration, purification, and filtration using the exosome concentration, purification, and filtration device of this utility model includes:

[0015] (1) Add the supernatant containing exosomes into the first filter bag through the feed pipe; (2) Turn on the stirrer in the first tank to allow the supernatant in the first filter bag to complete the permeation filtration; (3) Turn on the first peristaltic pump set on the first feed pipe to transport the filtrate after permeation filtration in the first filter bag to the second filter bag through the first feed pipe; concentrate the material in the second filter bag to obtain a concentrated liquid; (4) Turn on the second peristaltic pump set on the second feed pipe to transport the concentrated liquid from the second filter bag to the sterilization bottle through the second feed pipe.

[0016] In practical applications, the purification and concentration of exosomes are mainly achieved through filtration using filter bags. However, with prolonged use, contaminants can easily remain on the surface of these filter bags, affecting their filtration efficiency. This device uses multi-stage filter membranes to separate and purify exosomes. The first or second filter bag in this device can be replaced, significantly reducing costs and improving the efficiency of exosome acquisition. Attached Figure Description

[0017] Figure 1 Structural diagram of the exosome concentration, purification and filtration device of this utility model.

[0018] Figure 2 Nanoparticle tracking analysis (NTA) results - Malvern NS300.

[0019] Figure 3Transmission electron microscopy (TEM) results - Hitachi HT7700.

[0020] Figure 4 MSC exosome West Blot results (Marker: CD63 protein).

[0021] Figure 5 MSC exosome West Blot results (Marker: TSG101 protein).

[0022] 1-First tank, 2-Second tank, 3-Sterilization bottle, 4-Infeed pipe, 5-First filter bag, 6-Second filter bag, 7-First conveying pipe, 8-Second conveying pipe, 9-First peristaltic pump, 10-Second peristaltic pump, 11-Agitator, 12-Infeed pipe switch. Detailed Implementation

[0023] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer with the description. However, it should be understood that the embodiments described are merely exemplary and do not constitute any limitation on the scope of the present invention. Those skilled in the art should understand that modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but such modifications or substitutions all fall within the protection scope of the present invention.

[0024] This utility model provides an exosome concentration, purification, and filtration device, comprising a first tank 1, a second tank 2, a sterile bottle 3, an inlet pipe 4, a first filter bag 5, a second filter bag 6, a first conveying pipe 7, and a second conveying pipe 8; wherein, the first filter bag 5 is housed in the first tank 1, and the second filter bag 6 is housed in the second tank 2; the first filter bag 5 and the second filter bag 6 are connected through the first conveying pipe 7, and the second filter bag 6 and the sterile bottle 3 are connected through the second conveying pipe 8.

[0025] In a preferred embodiment of this utility model, a stirrer 11 is provided inside the first tank 1.

[0026] In a preferred embodiment of this utility model, a stirrer 11 is provided at the bottom of the first tank 1.

[0027] In a preferred embodiment of this utility model, a feed pipe 4 is provided on the first filter bag 5, and a feed pipe switch 12 is provided on the feed pipe 4; the material containing exosomes to be separated and purified is controlled to enter the first filter bag by the feed pipe and the feed pipe switch 12 provided thereon.

[0028] In a preferred embodiment of this utility model, a first peristaltic pump 9 is provided on the first conveying pipe 7. The material that has been filtered in the first filter bag 5 is pumped to the second filter bag 6 through the first conveying pipe 7 by the action of the first peristaltic pump 9.

[0029] In a preferred embodiment of this utility model, a second peristaltic pump 10 is provided on the second conveying pipe 8. The concentrated material in the second filter bag 6 is pumped to the sterilization bottle 3 through the second conveying pipe 8 by the action of the second peristaltic pump 10.

[0030] In a preferred embodiment of this utility model, the first filter bag 5 or the second filter bag 6 is separable from the first tank 1 or the second tank 2 or can be disassembled and removed from the first tank 1 or the second tank 2. Therefore, the used first filter bag 5 or the second filter bag 6 can be removed from the first tank 1 or the second tank 2 and replaced with a new first filter bag 5 or the second filter bag 6.

[0031] Example 1: Exosome concentration, purification, and filtration using the exosome concentration, purification, and filtration device of this invention.

[0032] Add 9 liters of physiological saline to the first tank 1; add 400 ml of supernatant containing exosomes to the first filter bag 5 through the feed pipe 4, close the feed pipe switch 12, turn on the stirrer 11, and stir for 5 hours; turn on the first peristaltic pump 9 to transfer the material in the first filter bag 5 to the second filter bag 6 through the first conveying pipe 7; add 2 liters of PEG6000 to the second tank 2, let it stand for 2 hours to concentrate the solution; turn on the second peristaltic pump 10 to transfer the concentrate in the second filter bag 6 to the sterile bottle 3 through the second conveying pipe 8. The sterile bottle 3 contains the concentrated and purified exosomes. Take 1 ml of the concentrated exosomes for testing, and obtain the test results as shown in Table 1. Figures 2-5 .

[0033] Table 1 Original Sample Information

[0034]

[0035] Table 2. Reference Standards for Exosome Quality Assessment

[0036]

[0037] The identification results show that the particle concentration is 9.61e+09partides / ml, the mode particle size is 146.6nm, and both CD63 and TSG101 bands are present in the Western blot analysis, which are consistent with the characteristics of exosomes.

Claims

1. An exosome concentration, purification, and filtration device, characterized in that, It consists of a first tank (1), a second tank (2), a sterile bottle (3), a feed pipe (4), a first filter bag (5), a second filter bag (6), a first conveying pipe (7), and a second conveying pipe (8); wherein, the first filter bag (5) is built into the first tank (1), and the second filter bag (6) is built into the second tank (2); the first filter bag (5) and the second filter bag (6) are connected through the first conveying pipe (7), and the second filter bag (6) and the sterile bottle (3) are connected through the second conveying pipe (8).

2. The exosome concentration, purification, and filtration apparatus according to claim 1, characterized in that, A stirrer (11) is installed inside the first tank (1).

3. The exosome concentration, purification, and filtration apparatus according to claim 2, characterized in that, A stirrer (11) is provided at the bottom of the first tank (1).

4. The exosome concentration, purification, and filtration apparatus according to claim 1, characterized in that, A feed pipe (4) is provided on the first filter bag (5), and a feed pipe switch (12) is provided on the feed pipe (4).

5. The exosome concentration, purification, and filtration apparatus according to claim 1, characterized in that, A first peristaltic pump (9) is installed on the first material conveying pipe (7).

6. The exosome concentration, purification, and filtration apparatus according to claim 1, characterized in that, A second peristaltic pump (10) is installed on the second material conveying pipe (8).

7. The exosome concentration, purification, and filtration apparatus according to claim 1, characterized in that, The first filter bag (5) or the second filter bag (6) is detachable from the first tank (1) or the second tank (2) or can be removed from the first tank (1) or the second tank (2) and replaced with a new filter bag.

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