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

Method for separating and subculturing primary neural stem cell of human embryo midbrain

A technology of primary cells and subculture, which is applied in the field of biomedicine and can solve the problem of less research on neural stem cells

Inactive Publication Date: 2013-04-03
陆华
View PDF0 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existence of neural stem cells in specific brain regions in mammalian embryonic stages and adult animals has been continuously confirmed, but there is little research on human neural 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
  • Method for separating and subculturing primary neural stem cell of human embryo midbrain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0013] Such as figure 1 Shown is a schematic flow chart of the steps of the method for the isolation and subculture of primary cells of human embryonic midbrain neural stem cells of the present invention, and the method is specifically realized by the following steps:

[0014] S1: Isolation and culture of primary midbrain neural stem cells: take fresh human embryos induced by water sac at 10-13 weeks of embryonic age, aseptically isolate the midbrain tissue of the embryos under a microscope, peel off the meninges and surface blood vessels, and put them in PBS ( Phosphate buffer solution) for three times, blow and beat the tissue fragments repeatedly with a fine pipette until they are completely dispersed, centrifuge and wash, blow and beat to make a cell suspension, filter through a 100-mesh stainless steel filter to make a single-cell suspension, add 1 :50 B27 (human leukocyte antigen), 20ng / ml EGF (epidermal growth factor), 20ng / ml bFGF (fibroblast growth factor) in DMEM / F12...

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 a method for separating and subculturing the primary neural stem cell of the human embryo midbrain. The method is used for separating, culturing and differentiating the primary neural stem cell of the human embryo midbrain in vitro by separating, culturing and identifying the neural stem cell of the human embryo midbrain. The method comprises the following steps: separating and culturing the primary neural stem cell of the human embryo midbrain, subculturing the primary cell, carrying out induced differentiation on the cell, carrying out immunocytochemical staining, and counting the number of cells. According to the method, a neural stem cell is separated from human embryo midbrain successfully by adopting the serum-free culturing and single-cell cloning technology, and the neural stem cell is cultured, subcultured and subjected to adherent differentiation observation, wherein the cell has the continuous cloning capacity and can subculture and express the neural nidogen antigen. The differentiated cell can express the specific antigens of the neuron cells, the colloid cells and oligodendroglia cells.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a method for separating and subcultureing primary cells of human embryonic midbrain neural stem cells, which is applied to the in vitro isolation, culture and differentiation process of human embryonic midbrain neural stem cells. Background technique [0002] The discovery of neural stem cells is a major breakthrough in the field of neuroscience, which has opened up broad prospects for research fields such as neural development and neural tissue transplantation. Neural stem cells are cells with self-renewal and extensive differentiation potential. They are in a non-terminal state of differentiation. They can divide symmetrically or asymmetrically into new stem cells or daughter cells with gradually decreasing differentiation potential, and finally produce the central nervous system. The three main types of cells are neurons, astrocytes, and oligodendrocytes. It is generally ...

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): C12N5/0797C12N5/0793
Inventor 陆华
Owner 陆华
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