A kind of cell culture medium and its application and method for inducing dental pulp stem cells to differentiate into nerve-like cells

A dental pulp stem cell and cell differentiation technology, applied in the direction of non-embryonic pluripotent stem cells, animal cells, nervous system cells, etc., can solve the problems of long cycle and low differentiation efficiency

Active Publication Date: 2018-12-04
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In vitro studies have found that DPSCs are pluripotent stem cells derived from neural crest and mesenchyme, and can differentiate into neural-like cells under specific induction conditions. lower

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 kind of cell culture medium and its application and method for inducing dental pulp stem cells to differentiate into nerve-like cells
  • A kind of cell culture medium and its application and method for inducing dental pulp stem cells to differentiate into nerve-like cells
  • A kind of cell culture medium and its application and method for inducing dental pulp stem cells to differentiate into nerve-like cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1～5

[0059] Using 3-5 generations of dental pulp stem cells, according to 2×10 3 Pcs / cm 2 The inoculation density is inoculated into a six-well plate. After culturing the cells with DMEM / F12 for 4h~6h to adhere to the wall, adding EGF respectively for pre-induction, discarding the complete medium, washing the cells twice with PBS, and adding the cell culture medium provided by the invention (including GDNF, Shh, EGF, cAMP) DMEM / F12 serum-free medium), after inducing cultured cells. Observe the differentiation of dental pulp stem cells. The pre-induction conditions, cell culture medium components and nuclear induction conditions used in each example are shown in Table 1:

[0060] Table 1, Examples 1 to 5

[0061]

[0062] After pre-induction and induction of the cells of each example, the cell morphology was observed with an electron microscope. The results showed that the cell morphology began to change at 12 hours, and marked changes occurred after 24 hours of induction. The refracti...

Embodiment 6

[0069] The total proteins induced by cells in Examples 1 to 5 and Comparative Examples 1 to 3 were extracted, and Western blot was performed to detect the changes in the expression of Nestin and MAP2 during the induction process. Specifically, the total proteins obtained from each extraction were subjected to 12% SDS-PAGE Polyacrylamide gel electrophoresis, 80V transfer membrane 1h to PVDF membrane. Block with 5% BSA at room temperature for 1 hour, add anti-Nestin, MAP2 primary antibody (1:200) or β-tubulin antibody (1:200), overnight at 4°C; wash the membrane, add HRP-labeled secondary antibody (1:2000), Incubate for 60min at room temperature; ECL glows. The result is Figure 5 .

[0070] The results showed that the expression levels of Nestin and MAP2 were increased after inducing the differentiation of dental pulp stem cells into nerve-like cells in each example and comparative example, which was consistent with the expression of specific proteins in nerve-like cells. The ex...

Embodiment 7

[0072] The cells were induced for 48 hours using the protocols of Comparative Example 1 and Example 1, and the cells were collected at 24 hours, 36 hours, and 48 hours of induction to extract total protein, and the expression levels of Nestin and MAP2 were detected by the method described in Example 6. The result is Image 6 . The results showed that after 24 hours of inducing cells in Example 1, the expression levels of Nestin and MAP2 in the cells were significantly higher than those in the induced cells in Comparative Example 1 for 48 hours.

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

PropertyMeasurementUnit
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
Login to view more

Abstract

The invention relates to the technical field of stem cell culture, in particular to cell culture fluid, application of the cell culture fluid and a method of inducting DPSCs (Dental Pulp Stem Cells) to differentiate into neuron-like cells. The cell culture fluid provided by the invention comprises basic culture fluid, GDNFs (Glial Cell-Derived Neurotrophic Factors), Shh (Sonic hedgehog), EGFs (Epidermal Growth Factors) and cAMP (Cyclic Adenosine monophosphate). The culture fluid provided by the invention can be used for inducting the DPSCs to differentiate into the neuron-like cells. When the culture fluid is used for inducing the DPSCs, the form of the DPSCs starts to change from 12h; after the 24h from the induction, the form of the DPSCs is obviously changed; the cell refractivity starts to be enhanced; cell processes increase and grows and differentiate into the form of the neuron-like cells. Through western bolt detection, after 24h from the induction, the expression amounts of Nestin and MAP2 (Microtubule-Associated Protein 2) in the DPSCs are obviously increased. Therefore the culture fluid has the advantages of short period and high efficiency when being used for inducing the DPSCs to differentiate into the neuron-like cells.

Description

Technical field [0001] The invention relates to the technical field of stem cell culture, in particular to a cell culture solution and its application, and a method for inducing dental pulp stem cells to differentiate into nerve-like cells. Background technique [0002] Many clinically common chronic neurodegenerative diseases, such as Parkinson’s syndrome, Alzheimer’s disease, Huntington’s disease and amyotrophic lateral sclerosis. These diseases are caused by the degeneration and death of nerve cells in the central nervous system. In addition, external trauma can also cause nerve damage, leading to severe behavioral dysfunction. [0003] Nerve cells will not regenerate as quickly as epidermis and liver cells. Although stem cells in the central nervous system can also replicate and differentiate, their replication speed is extremely slow, and the proportion of differentiated into nerve cells is also very low. Most of them will differentiate into glial cells, which cannot be trea...

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): C12N5/074C12N5/079
Inventor 陈海佳王一飞葛啸虎冯德龙马岩岩
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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