Check patentability & draft patents in minutes with Patsnap Eureka AI!

Extraction method of exosome

An extraction method and technology for exosomes, which are applied in cell dissociation methods, biochemical equipment and methods, artificial cell constructs, etc., can solve problems such as insufficient extraction of exosomes, and achieve low cost, few steps, and simple steps. Effect

Pending Publication Date: 2021-12-28
深圳协同创新高科技发展有限公司 +1
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the embodiments of the present invention is to provide a method for extracting exosomes, which aims to solve the problem of insufficient extraction of exosomes by ultracentrifugation, filter centrifugation, and immunomagnetic bead methods currently used in the prior art

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Extraction method of exosome
  • Extraction method of exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for extracting exosomes, the extraction method comprising the following steps:

[0022] 1) Preparation of PEG extract: Take polyethylene glycol with a molecular weight of 4000 and a quality of 10g and 29.25g of sodium chloride and fully dissolve it in pure water to obtain an extract. The concentration of polyethylene glycol in the extract is 0.1g / L, the concentration of sodium chloride is 0.5mol / L;

[0023] 2) Cell culture: when the cell confluency is about 60%, remove the original serum-containing medium, rinse the cells three times with phosphate buffer solution with a pH of 7.4, and add fresh medium without serum-related components to carry out Cultivate, continue to cultivate, collect the cell culture supernatant when the cell confluence reaches 90%, collect the culture medium, centrifuge, remove cell debris, and obtain the initial supernatant;

[0024] 3) Exosome extraction: centrifuge the initial supernatant in step 2) at 300g centrifugal force, 4°C for...

Embodiment 2

[0026] A method for extracting exosomes, the extraction method comprising the following steps:

[0027] 1) Preparation of PEG extract: Take polyethylene glycol with a molecular weight of 5000 and a quality of 20g and 58.5g of sodium chloride and fully dissolve it in pure water to obtain an extract. The concentration of polyethylene glycol in the extract is 0.2g / L, the concentration of sodium chloride is 1mol / L;

[0028] 2) Cell culture: when the cell confluency is about 65%, remove the original serum-containing medium, rinse the cells three times with phosphate buffer solution with a pH of 7.4, and add fresh medium without serum-related components to carry out Cultivate, continue to cultivate, collect the cell culture supernatant when the cell confluence reaches 90%, collect the culture medium, centrifuge, remove cell debris, and obtain the initial supernatant;

[0029] 3) Exosome extraction: centrifuge the initial supernatant in step 2) at 500g centrifugal force, 20°C for 8...

Embodiment 3

[0031] A method for extracting exosomes, the extraction method comprising the following steps:

[0032] 1) Preparation of PEG extract: Take polyethylene glycol with a molecular weight of 6000 and a quality of 30g and 87.75g of sodium chloride and fully dissolve it in pure water to obtain an extract. The concentration of polyethylene glycol in the extract is 0.3g / L, the concentration of sodium chloride is 1.5mol / L;

[0033] 2) Cell culture: when the cell confluency is about 70%, remove the original serum-containing medium, rinse the cells three times with phosphate buffer solution with a pH of 7.4, and add fresh medium without serum-related components to carry out Cultivate, continue to cultivate, collect the cell culture supernatant when the cell confluence reaches 90%, collect the culture medium, centrifuge, remove cell debris, and obtain the initial supernatant;

[0034] 3) Exosome extraction: centrifuge the initial supernatant in step 2) at 600g centrifugal force, 30°C fo...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to the technical field of extraction processes, and provides an extraction method of an exosome. The method comprises the following steps of (1) preparation of a PEG extracting solution: weighing polyethylene glycol and sodium chloride, and fully dissolving the polyethylene glycol and the sodium chloride in pure water to obtain the extracting solution; and (2) cell culture: when the cell fusion degree is about 60-70%, removing an original serum-containing culture medium, rinsing cells for three times by using a phosphate buffer solution, adding a fresh serum-free related component culture medium for culture, continuing culture until the cell fusion degree reaches 90%, collecting cell culture supernatant, collecting the culture medium, centrifuging, removing cell debris, and obtaining an initial supernatant; and (3) exosome extraction: carrying out centrifugation treatment on the initial supernatant obtained in the step (2) twice, taking the supernatant, adding the extract obtained in the step (1), carrying out inversion, fully mixing, incubating, carrying out centrifugation treatment, discarding the supernatant, collecting precipitate, suspending with the phosphate buffer, and storing a sample subgroup at-80 DEG C. The method is simple in step, low in cost and capable of achieving mass production.

Description

technical field [0001] The invention belongs to the technical field of extraction technology, and in particular relates to an extraction method of exosomes. Background technique [0002] Exosomes are a kind of extracellular vesicles (Extracellular vesicles, EVs) secreted by living cells, with a diameter of about 30-200nm and a density of 1.13-1.21g / ml, with a typical lipid bilayer structure , which is released outside the cell in the form of exocrine after the fusion of multivesicular bodies (MVBs) in the cell and the cell membrane. Almost all types of cells (immune cells, nerve cells, stem cells), including tumor cells, can produce and release exosomes, but the RNA, lipids, and protein components contained in exosomes from different cell sources are not the same . The functional proteins, mRNA and microRNA contained in exosomes play an important role in the process of information transmission between cells, participate in the transportation of substances and information t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0602C12N2509/00
Inventor 周旋周忠娇王艺璇李昕李娜娜
Owner 深圳协同创新高科技发展有限公司
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com