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

Preparation method of neural stem cell

A technology of neural stem cells and plasmic stem cells, which is used in nervous system cells, animal cells, nervous system diseases, etc.

Active Publication Date: 2015-03-18
微能生命科技集团有限公司
View PDF3 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of individual development, there are various forms of stem cells, such as embryonic stem cells, pluripotent stem cells, and committed stem cells in various tissues, such as hematopoietic stem cells, neural stem cells, etc., but they face ethical challenges or limitations in practical application. limit

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
  • Preparation method of neural stem cell
  • Preparation method of neural stem cell
  • Preparation method of neural stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Embodiment 1, the preparation of culture medium

[0087] Preparation of medium A, B, C, D and E:

[0088] The first medium (medium A) consists of animal cell basal medium and solutes. Animal cell basal medium is Knockout DMEM or Knockout DMEM / F12; solutes and their concentrations in animal cell basal medium are as follows: 200ml / L Knockout serum substitute, 10ml / L non-essential amino acids, 0.1mmol / L β-mercaptoethanol , 1mmol / L L-glutamine, 4ng / ml b-FGF.

[0089] The second medium (medium B) consists of animal cell basal medium and solutes. The animal cell basal medium is Neurobasal medium and DMEM / F12 medium with a volume ratio of 1:1; the solute and its concentration in the animal cell basal medium are as follows: 10ml / L N2 supplement, 20ml / L B27 supplement, 1mmol / L L-glutamine, 0.1mmol / Lβ-mercaptoethanol.

[0090] The third medium (medium C) consists of animal cell basal medium and solutes. The animal cell basal medium is Neurobasal medium and DMEM / F12 medium w...

Embodiment 2

[0093]Example 2, Induced Culture of Human Neural Stem Cells

[0094] Step 1. Obtain the third generation of mesenchymal stem cells

[0095] 1. Wash the adipose tissue collected by liposuction with D-Hank’s (or PBS) to remove blood cells and anesthetics, digest with 2g / L type II collagenase at 37°C for 30min, filter through a 100-mesh sieve and collect the filtrate.

[0096] 2. Resuspend the cells obtained in step 1 with D-Hank's solution and wash twice to remove collagenase, centrifuge at 1200rpm at room temperature for 10min, and collect the cells.

[0097] 3. Divide the cells from step 2 in a 2×10 6 The density of each / ml was inoculated in the expansion culture medium (an example of a formula is containing 580ml / L DMEM / F12, 400ml / L MCDB, 20ml / L fetal bovine serum, 1×10 -9 mol / L insulin-transferrin-selenium supplement (ITS), 1×10 -9 mol / L dexamethasone, 1×10 -4 mol / L 2-phosphate ascorbic acid, 20ng / mL interleukin-6, 10ng / mL EGF, 10ng / mL PDGF-BB, 100U / mL penicillin and 100...

Embodiment 3

[0107] Example 3, identification of the terminal differentiation ability of the obtained neural stem cells

[0108] 1. Differentiate into neurons: the cells finally obtained in step 2 of Example 2 were planted in a 6-well plate coated with polyguanine / laminin (PDL / laminin), 5×10 per well 5 Add the fourth medium (ie medium D) to the cells, change the medium every 2 days, and co-culture for 12-16 days, such as 14 days.

[0109] 2. Differentiation to glia: the cells finally obtained in step 2 of Example 2 were planted in a 6-well plate coated with polyguanine / laminin (PDL / laminin), 5×10 per well 5 Add medium E to the cells, change the medium every 2 days, and co-culture for 12-16 days, such as 14 days.

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 relates to a preparation method of a neural stem cell and particularly discloses a method for inducing a human mesenchymal stem cell to obtain the neutral stem cell. The method comprises the step of sequentially cultivating the mesenchymal stem cell in first, second and third culture media to obtain the neural stem cell. The finally obtained neural stem cell expresses a mark of an early neural stem cell and can be differentiated to functional neuron and neurogliocyte.

Description

technical field [0001] The present invention relates to the field of regenerative medicine. Specifically, it relates to a preparation method of neural stem cells. More specifically, the present invention relates to a method for inducing mesenchymal stem cells into neural stem cells. Background technique [0002] The incidence of degenerative diseases of the nervous system is increasing year by year, mainly manifested as abnormal function of nerve cells or progressive loss of cells, and it is a common clinical refractory disease. For the body itself, dysfunctional nerve cells are difficult to repair by itself, and the limited nerve regeneration ability of the body itself cannot meet the demand for the number of cells needed to repair damage or replace cells. At present, for these refractory diseases, cell nutrition therapy or drug therapy is generally used clinically, but the effect is minimal, and there is no safe and effective treatment method yet. Therefore, the treatme...

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): C12N5/0797C12N5/0793C12N5/079A61K35/30A61P25/00A61P25/16A61P25/28
Inventor 赵春华
Owner 微能生命科技集团有限公司
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