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Method for increasing content of exosomes released by cells

An exosome content technology, applied in the direction of artificial cell constructs, gastrointestinal cells, animal cells, etc., can solve the problems of high cost, poor applicability of different cells, complicated operation, etc., and achieve the goal of increasing the release of exosomes Effect

Inactive Publication Date: 2022-04-22
多莱泌生物科技(武汉)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Exosome-based biomedical treatment technology is becoming more and more mature. For example, using the vesicle structure of exosomes to encapsulate therapeutic drugs can directly target the lesion to treat diseases. Compared with the current widespread use of liposome drug delivery, exosome drug delivery has It has the advantages of good biocompatibility, strong stability, low immune rejection and easy absorption by cells. However, the extraction conditions of exosomes are harsh and the cost is high. Therefore, a simple, fast and low-cost method to stimulate the production of exosomes Has important socioeconomic significance and clinical value
[0004] Exosomes are a hot spot in new science in recent years. There are still many mechanisms in the secretion process that are still unclear. Regulating the secretion of exosomes in cells is a complex process. At present, the release of exosomes in cells cannot be improved by changing gene expression. Quantitative methods include low-intensity pulsed ultrasonic stimulation, different LED light irradiation stimulation methods, and special medium treatment methods, but the above methods are complicated to operate and have poor applicability to different cells. body content method to solve the above problems

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  • Method for increasing content of exosomes released by cells
  • Method for increasing content of exosomes released by cells
  • Method for increasing content of exosomes released by cells

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Experimental program
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Effect test

Embodiment 1

[0065] see figure 1 , the method for increasing the content of exosomes released by cells in the present invention, comprising the following steps:

[0066] 1) Liver cancer cells were replaced with fresh medium;

[0067]2) Liver cancer cells are irradiated with UVB for 1 hour, and the status of liver cancer cells is always paid attention to;

[0068] 3) Liver cancer cells were cultured in an incubator for 2 hours;

[0069] 4) Place the liver cancer cells in the hypoxic device for 1 hour;

[0070] 5) Collect the liver cancer cell supernatant after 24 hours;

[0071] 6) Filter the treated liver cancer cell supernatant with a 0.45um filter membrane, and collect the filtered liquid;

[0072] 7) Centrifuge at 120000g for 90min to extract exosomes;

[0073] 8) Collect exosomes;

[0074] 9) Carry out various tests.

Embodiment 2

[0076] see figure 1 , the method for improving the content of exosomes released by cells in the present invention, comprising the following steps:

[0077] 1) Colorectal cancer cells were replaced with fresh medium;

[0078] 2) Colorectal cancer cells are irradiated with UVB for 1 hour, and the status of colorectal cancer cells is always concerned;

[0079] 3) Colorectal cancer cells were cultured in an incubator for 2 hours;

[0080] 4) Place the colorectal cancer cells in the hypoxic device for 1 hour;

[0081] 5) Collect the colorectal cancer cell supernatant after 24 hours;

[0082] 6) Filter the treated colorectal cancer cell supernatant with a 0.45um filter membrane, and collect the filtered liquid;

[0083] 7) Centrifuge at 120000g for 90min to extract exosomes;

[0084] 8) Collect exosomes;

[0085] 9) Carry out various tests.

Embodiment 3

[0087] see figure 1 , the method for improving the content of exosomes released by cells in the present invention, comprising the following steps:

[0088] 1) Oral cancer cells were replaced with fresh medium;

[0089] 2) Oral cancer cells are irradiated with UVB for 1 hour, and the status of oral cancer cells is always concerned;

[0090] 3) Oral cancer cells were cultured in an incubator for 2 hours;

[0091] 4) Oral cancer cells were placed in the hypoxic device for 1 hour;

[0092] 5) Collect the oral cancer cell supernatant after 24 hours;

[0093] 6) Filter the treated oral cancer cell supernatant with a 0.45um filter membrane, and collect the filtered liquid;

[0094] 7) Centrifuge at 120000g for 90min to extract exosomes;

[0095] 8) Collect exosomes;

[0096] 9) Carry out various tests.

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Abstract

The invention relates to the technical field of engineered exosomes, and discloses a method for increasing the content of exosomes released by cells, which comprises the following steps: 1) treating a cancer cell line; 2) filtering the treated cancer cell supernatant with a 0.45 [mu] m filter membrane, and collecting the filtered liquid; (3) centrifuging 120000g of the solution for 90 minutes, and extracting the exosome; 4) collecting the exosome; the cancer cell line treatment comprises the following steps: 1) replacing a fresh culture medium for cancer cells; 2) irradiating cancer cells with UVB for 1 hour, and paying attention to the state of liver cancer cells at any moment; 3) culturing the cancer cells in an incubator for 2 hours; 4) placing the cancer cells in an anoxic device for 1 h; and 5) collecting the supernatant of the cancer cells after 24 hours. The exosome release amount can be increased under the condition that cell genetic materials are not changed, the method is suitable for various cell lines, compared with an ultrasonic method, an LED lamp irradiation method, a special culture medium treatment method and other methods, the method is simpler, and the exosome release amount of the various cell lines can be better increased.

Description

technical field [0001] The invention relates to the technical field of engineered exosomes, in particular to a method for increasing the content of exosomes released by cells. Background technique [0002] Exosomes are tiny vesicles with a lipid bilayer membrane structure, with a diameter of 30-150nm, which can be secreted by almost all cells and can mediate material exchange and information exchange between cells. Studies have shown that exosomes carry many In addition, exosomes have immunogenic properties, come from a wide range of sources, and can carry RNA, protein, DNA, and small molecules. [0003] Exosome-based biomedical treatment technology is becoming more and more mature. For example, using the vesicle structure of exosomes to encapsulate therapeutic drugs can directly target the lesion to treat diseases. Compared with the current widespread use of liposome drug delivery, exosome drug delivery has It has the advantages of good biocompatibility, strong stability, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071C12N13/00
CPCC12N5/0693C12N5/067C12N5/0679C12N5/0632C12N13/00C12N2500/02C12N2529/10
Inventor 李立吴笛笛刘瑾陈佳佳高强刘丕菊
Owner 多莱泌生物科技(武汉)有限公司
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