A culture method for maintaining the self-renewal state of human embryonic stem cells

A technology for human embryonic stem cells and embryonic stem cells, applied in the field of embryonic stem cell culture conditions, can solve problems such as unfavorable exploration, unstable state of human embryonic stem cells, and inconvenient passage operations.

Active Publication Date: 2019-11-22
ANHUI UNIVERSITY
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of unstable state of human embryonic stem cells, inconvenient subculture operation, high cost of existing culture conditions and unfavorable follow-up exploration in traditional culture conditions, the present invention provides a culture method for maintaining the self-renewal state of human embryonic 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
  • A culture method for maintaining the self-renewal state of human embryonic stem cells
  • A culture method for maintaining the self-renewal state of human embryonic stem cells
  • A culture method for maintaining the self-renewal state of human embryonic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] (1) HES2 human embryonic stem cells (Human Embryonic Stem Cells, hESCs) were provided by Professor Qilong Ying from the University of Southern California;

[0071] (2) Passage and culture of HES2 human embryonic stem cells under the condition of Activin A+BFGF+IWR1:

[0072] a. Take a 6-well cell culture plate, wrap each well with 2ml DMEM+10μl Matrigel, and place at 37°C, 5% CO 2 Concentration in the cell culture box, coated for 4 hours;

[0073] b. Take the human embryonic stem cells grown to 70-80% density, discard the culture medium, and wash the cells once with PBS to remove the residual culture medium on the cell surface;

[0074] c. Add 1ml of CTK to digest the embryonic stem cells. After about 7 minutes, the edge of the cells floats. Discard the CTK, add 2ml of conventional cell culture medium containing 10% FBS, blow and blow the cells from the culture plate with a pipette, and then transfer into a 15ml sterile centrifuge tube;

[0075] d. After centrifugati...

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 culturing method for maintaining a self-renewal state of human embryonic stem cells. The method comprises passage and culturing of the human embryonic stem cells under the condition of using the culturing method and observation and detection of the self-renewal state of the human embryonic stem cells cultured under the condition of using the culturing method. According tothe method, the problems of easily differentiated state, complicated passage operation, high culturing condition cost and inconvenient subsequent research on mechanisms of the human embryonic stem cells under the current traditional culturing condition are solved by adding an inhibitor of a Wnt / beta-catenin signal channel; the method has the advantages of being stable in effect, economic and high-efficient, simple to operate, convenient to research and the like, clues for improving and establishing culturing conditions for multipotential stem cells of other kinds and species are provided, and the method has a positive promotion effect on smooth development of stem cell basis and application research in the future.

Description

technical field [0001] The present invention relates to culture conditions of human embryonic stem cells, in particular to a culture method for maintaining the self-renewal state of human embryonic stem cells. Background technique [0002] Stem cells are self-renewing pluripotent cells that are isolated from early embryos and can survive in vitro culture conditions and have the ability to proliferate indefinitely (self-renewal), while maintaining the ability to differentiate into various types of cells in the body Its potential (differentiation) has become one of the powerful tools for studying gene function, screening drugs and making animal models of diseases, and has great application prospects in the field of regenerative medicine. Mouse Embryonic StemCells (mESCs), which is the first stem cell line successfully established in vitro. Later, in 1998, Thomson, the "father of stem cells", established the human ESC line for the first time. Although mouse and human ESCs sha...

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): C12N5/0735
CPCC12N5/0606C12N2501/115C12N2501/16C12N2501/999
Inventor 叶守东尤宇
Owner ANHUI UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap