Culture method for promoting proliferation and differentiation of neural stem cells

A neural stem cell and culture method technology, applied in nervous system cells, cell culture active agents, biochemical equipment and methods, etc., can solve problems such as unsatisfactory effects, and achieve the effect of promoting differentiation and proliferation in the direction of neurons

Inactive Publication Date: 2017-02-15
青海七彩花生物科技有限公司
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Researchers tried to adjust the proliferation and differentiation of NSCs by supplementing exogenous

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
  • Culture method for promoting proliferation and differentiation of neural stem cells
  • Culture method for promoting proliferation and differentiation of neural stem cells
  • Culture method for promoting proliferation and differentiation of neural stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of exogenous neurotrophic factors and strengthening peptides

[0026] The synthesis method of peptides is very mature. It can be synthesized by solid-phase method in the laboratory itself, or can be directly entrusted to a biological outsourcing company for synthesis. The polypeptides involved in the present invention are all synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and identified by ESI mass spectrometry to ensure the correct sequence of the polypeptides.

[0027] Fmoc method solid-phase synthesis technology: the synthesis reaction is carried out from the C-terminal to the N-terminal, and there are free amino groups on the Rink medium; during each step of the connection process, the amino acid residues must be activated, and the activation mixture has 4 times the number of free amino groups on the medium HBTU, HOBt, DIEA and Fmoc-amino acids; after each amino acid linking reaction, a mixture of pyridine / acetic acid / N-methylimida...

Embodiment 2

[0029] Example 2: Isolation, culture and identification of neural stem cells

[0030] The isolation and culture method of primary neural stem cells is as follows: 14 days pregnant mice were taken, killed by cervical dislocation, placed in 75% ethanol for 5 minutes, the pregnant uterus was taken out, rinsed twice with pre-cooled D-hank's, the cerebral cortex was peeled off after the embryos were taken out, Place in pre-cooled D-hank's solution, cut into pieces with ophthalmic scissors and blow repeatedly to make a cell suspension, filter through a sterile 200-mesh steel mesh and centrifuge at 1500r / min for 5min, regenerate with DMEM / F12 (1:1) medium Suspended cells, pipetting again, counting, 2×10 5 cells / mL were inoculated in modified DMEM / F12 (1:1) medium at 37°C, 5% CO 2 and 95% air conditions for 2-3 days, the neural stem cells will form primary neurospheres, continue to culture for 2-3 days, use 0.25% trypsin and blow the neurospheres mechanically to make a single cell su...

Embodiment 3

[0033] Example 3: Effects of Exogenous Neurotrophic Factors on NSCs Proliferation

[0034] The third-generation neural stem cells isolated and cultured in vitro were digested and pipetted into single cells, and seeded in a single layer into a 12-well culture plate with built-in polylysine-treated cell slides. The cultured neural stem cells were divided into control group and drug-treated group, at 37°C, 5% CO 2 Cultivate under 95% air condition, add neural stem cell serum-free medium, add BrdU for 48 hours, the final concentration of BrdU is 20ng / L, continue to cultivate for 2 hours, collect cell slides, and perform BrdU staining, the primary antibody dilution is 1:200, DAPI was added dropwise to stain the nucleus, the number of BrdU-positive cells and the total number of cells were counted, and the ratio of BrdU-positive cells was calculated; wherein, the serum-free medium for neural stem cells was DMEM / F12 (1:1) medium. The serum-free medium of neural stem cells in the drug...

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 culture method for promoting proliferation and differentiation of neural stem cells. According to the culture method, exogenous neurotrophic factors act to the cultured neural stem cells, so that the neural stem cells are induced to have proliferation and differentiation, and then neurons are formed, wherein the exogenous neurotrophic factors belong to a polypeptide with the sequence shown as SEQ ID NO.1; the exogenous neurotrophic factors and enhanced peptide act to the cultured neural stem cells together, so that the neural stem cells are induced to have proliferation and differentiation, and then neurons are formed, wherein the enhanced peptide is an oligopeptide with the sequence shown as SEQ ID NO.2, SEQ ID NO.3 or SEQ ID NO.4. According to the method provided by the invention, by supplementing the exogenous neurotrophic factors, the proliferation of neural stem cells and the differentiation of the neural stem cells towards the neurons can be obviously promoted, the enhanced peptide can enhance the effects of promoting proliferation and inducing differentiation of the exogenous neurotrophic factors, and the effects are obvious.

Description

technical field [0001] The invention belongs to the biological field and relates to the cultivation of stem cells, in particular to a culture method for promoting the proliferation and differentiation of neural stem cells, which is used for promoting the proliferation and differentiation of neural stem cells towards neurons. Background technique [0002] Neural stem cells (Neural Stem Cells, NSCs) are a kind of stem cells, which are undifferentiated cells existing in the brain and spinal cord. Like other stem cells, neural stem cells also have the points of division, proliferation and diverse differentiation, and have the ability of self-renewal. It can differentiate into neurons, glial cells and other types of nerve tissue cells. The various functions of neural stem cells are completed by neurons and glial cells. Many diseases in the central nervous system, such as: brain injury, cerebral hemorrhage, Parkinson's disease, etc., have varying degrees of loss of nerve cells or...

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): C12N5/0793C12N5/0797
CPCC12N5/0619C12N2501/13C12N2506/08
Inventor 杨廷伟刘翠红崔长年陈四海朱荣富赵元英朱晓翔谷鹏飞陈子扬邓凯旋
Owner 青海七彩花生物科技有限公司
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