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

Enrichment and Screening Methods of Cancer Stem Cells

A technology of tumor stem cells and screening methods, which is applied in the direction of tumor/cancer cells, biochemical equipment and methods, animal cells, etc., can solve the problems of difficult sorting and enrichment of tumor stem cells, and achieve high yield and high purity of stem cells Effect

Active Publication Date: 2020-03-27
湖南艾佳生物科技股份有限公司
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] The purpose of the present invention is to provide a new compound tumor stem cell screening method to solve the problem that it is difficult to effectively sort and enrich tumor stem cells

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
  • Enrichment and Screening Methods of Cancer Stem Cells
  • Enrichment and Screening Methods of Cancer Stem Cells
  • Enrichment and Screening Methods of Cancer Stem Cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Combined sorting of renal cancer cell line ACHN tumor stem cells (CD133 / CD24 positive double selection)

[0054] 1. Multi-generation sphere culture

[0055] 1) Cultivate ACHN tumor cells until the bottle is 80-90% full, digest and collect the cells, and the single cell suspension is 5×10 4 cells / ml were inoculated into 6-well ultra-low adsorption cell culture plates, and each well was cultured with 2 mL of SFMD medium;

[0056] 2) 37°C, 5% CO 2 Cultivate for 96 hours, add 1 mL of culture medium every other day;

[0057] 3) After 96 hours, at 37°C, 5% CO 2 Digest with 0.05% trypsin in the cell culture incubator for 5 minutes, the tumor spheres become loose, add 2 mL of SFMD medium to stop digestion, pipette evenly, take 10 μL of uniform cell suspension for cell counting, continue to press 5×10 4 Cells per ml were planted in a new 6-well ultra-low adsorption cell culture plate to start P2 generation sphere culture;

[0058] 4) Repeat the above steps 1) to 3) ...

Embodiment 2

[0130] Example 2 Joint sorting of bladder cancer cell line 5637 tumor stem cells (CD133 / CD44 positive double selection)

[0131] 1. Multi-generation sphere culture

[0132] 1) Cultivate 5637 tumor cells until the bottle is 80-90% full, digest and collect the cells, and press 5×10 single-cell suspension 4 cells / ml were inoculated into 6-well ultra-low adsorption cell culture plates, and each well was cultured with 2 mL of SFMD medium;

[0133] 2) 37°C, 5% CO 2 Cultivate for 96 hours, add 1mL culture medium every other day;

[0134] 3) After 96 hours, at 37°C, 5% CO 2 Digest with 0.05% trypsin in the cell culture incubator for 5 minutes, the tumor spheres become loose, add 2 mL of SFMD medium to stop digestion, pipette evenly, take 10 μL of uniform cell suspension for cell counting, continue to press 5×10 4 Cells per ml were planted in a new 6-well ultra-low adsorption cell culture plate to start P2 generation sphere culture;

[0135] 4) Repeat the above steps 1) to 3) to c...

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 provides an enriching and screening method for tumor stem cell. The combination of ball culture and immunomagnetic beads separation method is used for enriching and screening the tumor stem cells, the details are as follows: continuously culturing the tumor cells in a no-serum ball culture medium, using the QPCR method to simultaneously examine the relative levels of tumor stem cell markers, when the expression levels of specific marker is relatively the highest, supplement with immunomagnetic and positive, single or double screening tumor stem cells. The combination of the two screening methods can significantly improve such practical problems as low separation rates, low purity levels and the inconveniences of after application for tumor stem cells.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for enriching and screening tumor stem cells. Background technique [0002] The traditional concept holds that tumors are formed by somatic cell mutations, each tumor cell can grow indefinitely, and the occurrence and development of tumors is the result of the joint proliferation of all tumor cells. But this cannot explain the phenomenon that not all tumor cells can grow unrestricted. The characteristics of tumor cell growth, metastasis and recurrence are very similar to the basic characteristics of stem cells. Therefore, some scholars put forward the theory of cancer stem cells (CSC). This theory provides a new direction and perspective for people to re-understand the origin and nature of tumors, as well as clinical tumor treatment. [0003] Recent studies have confirmed that there are a small number of stem cell-like cell populations in tumor tissues. These cells have sel...

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 Patents(China)
IPC IPC(8): C12N5/095
CPCC12N5/0695C12N2500/32C12N2501/11C12N2501/115C12N2501/33C12N2501/998
Inventor 艾立平魏薇胡清云
Owner 湖南艾佳生物科技股份有限公司
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