Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Purification and concentration preparation method of stem cell exocrine

An exocrine and stem cell technology, applied in cell dissociation methods, biochemical equipment and methods, animal cells, etc., can solve the problems of affecting the biological activity of exosomes, low extraction concentration, etc., to achieve high purity and recovery rate, high recovery rate , the effect of simple operation

Inactive Publication Date: 2020-12-01
广州北斗生物科技有限公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extraction process of the existing technology affects the biological activity of exosomes, and the extraction concentration is low. Therefore, we propose a method for the purification and concentration of stem cell exosomes

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

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0015] The present invention provides a technical solution: a preparation method for purification and concentration of stem cell exocrine bodies, comprising the following steps:

[0016] S1. Quickly filter the cell culture solution with a 0.22 μm filter sieve, separate the complete cells and residues, and use a centrifuge to ultracentrifuge at a speed of 10000-11500g for 2 hours;

[0017] S2. Resuspend and wash the small vesicles with 1 mL of phosphate buffered saline at room temperature, and then use a centrifuge to ultracentrifuge at a speed of 10000-11500 g for 2 hours;

[0018] S3. Resuspend with 100 μL normal temperature phosphate-buffered saline solution and transfer to a low-adhesion tube to obtain high-concentration exosomes.

[0019] This technical solution purifies exocrine bodies through multiple ultracentrifugation or ultrafiltration centrifugation. The operation is simple, the number of vesicles obtained is large, and the biological activity of exocrine bodies is ...

Embodiment 1

[0021] S1. Quickly filter the cell culture solution with a 0.22 μm filter sieve, separate the complete cells and residues, and use a centrifuge to ultracentrifuge at a speed of 10000-11500g for 2 hours;

[0022] S2. Resuspend and wash the small vesicles with 1 mL of phosphate buffered saline at room temperature, and then use a centrifuge to ultracentrifuge at a speed of 10000-11500 g for 2 hours;

[0023] S3. Resuspend with 100 μL normal temperature phosphate-buffered saline solution and transfer to a low-adhesion tube to obtain high-concentration exosomes.

[0024] This technical solution purifies exocrine bodies through multiple ultracentrifugation or ultrafiltration centrifugation. The operation is simple, the number of vesicles obtained is large, and the biological activity of exocrine bodies is not affected. High-purity and high-recovery exocrine bodies can be obtained to solve the problem. The problem of affecting the biological activity of exosomes and low extraction co...

Embodiment 2

[0026] S1. Quickly filter the cell culture solution with a 0.22μm filter screen to separate the complete cells and residues, and use a centrifuge to ultracentrifuge at a speed of 10000-11500g for 2 hours. The cell culture solution is 80%-90% from sterile The cultured cells were mixed with protease inhibitors at a concentration ratio of 1:1.000;

[0027] S2. Resuspend and wash the small vesicles with 1 mL of phosphate buffered saline at room temperature, and then use a centrifuge to ultracentrifuge at a speed of 10000-11500 g for 2 hours;

[0028] S3. Resuspend with 100 μL normal temperature phosphate-buffered saline solution and transfer to a low-adhesion tube to obtain high-concentration exosomes.

[0029] This technical solution purifies exocrine bodies through multiple ultracentrifugation or ultrafiltration centrifugation. The operation is simple, the number of vesicles obtained is large, and the biological activity of exocrine bodies is not affected. High-purity and high-r...

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

No PUM Login to View More

Abstract

The invention discloses a purification and concentration preparation method of stem cell exocrine. The method comprises the following steps: S1, quickly filtering a cell culture solution with a 0.22 [mu]m filter sieve, separating complete cells and residues, and performing ultracentrifugation for 2 hours at a speed of 10000-11500 g with a centrifuge; S2, re-suspending and cleaning small vesicles with 1 mL of a normal-temperature phosphate buffered saline, and performing ultracentrifugation for 2 hours at a speed of 10000-11500 g with the centrifuge again; and S3, performing re-suspending with100 [mu]L of the normal-temperature phosphate buffered saline and then performing transferring into a low-adhesion tube to obtain the high-concentration exocrine, wherein the cell culture solution isobtained from 80%-90% of sterile cultured cells. According to the method, the exocrine is purified through multiple times of ultracentrifugation, the operation is simple, the number of obtained vesicles is relatively large, the biological activity of the exocrine is not affected, the exocrine with high purity and high recovery rate can be obtained, and problems that in the extraction process of the prior art, the biological activity of the exocrine is affected, and the extraction concentration is low are solved.

Description

technical field [0001] The invention relates to the technical field of exocrine extraction, in particular to a preparation method for purification and concentration of stem cell exocrine. Background technique [0002] Discovered in 1986, exosomes are double-membrane vesicles with a diameter of about 30nm-100nm, which can be produced by various cells in the body such as immune cells, stem cells, cardiovascular cells, reticulocytes, platelets, and nerve cells. It is actively secreted by tumor cells and is widely distributed in peripheral blood, urine, saliva, milk, ascites, amniotic fluid and other body fluids. [0003] Exosomes carry a large number of specific proteins (such as cytokines, growth factors) and functional biologically active substances, which participate in physiological processes such as cell communication, cell migration, angiogenesis, and anti-tumor immunity in the body, and are associated with the occurrence of various diseases closely related to the proces...

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
IPC IPC(8): C12N5/07
CPCC12N5/06C12N2509/10
Inventor 朱瑜
Owner 广州北斗生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com