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Nervous network restoration system, as well as preparation method and application thereof

A technology for repairing systems and neural networks, applied to nervous system diseases, biochemical equipment and methods, applications, etc., can solve the problems of lack of cell specificity, time controllability, poor curative effect, etc., to overcome transient release, Effects of promoting proliferation and repairing damaged neural networks

Active Publication Date: 2014-02-19
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method of using electrical stimulation or drugs lacks cell specificity and time controllability in vivo, because both electrical stimulation and drug effects are often released transiently, and the curative effect is poor

Method used

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  • Nervous network restoration system, as well as preparation method and application thereof
  • Nervous network restoration system, as well as preparation method and application thereof
  • Nervous network restoration system, as well as preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0068] see figure 1 , a method for preparing a neural network repair system according to an embodiment includes the following steps S110 to S150.

[0069] Step S110: constructing photosensitive astrocytes.

[0070] Photosensitive astrocytes are jointly constructed by astrocytes, photosensitive genes and viral vectors carrying photosensitive genes.

[0071] The astrocytes are preferably animal hippocampal astrocytes or striatal astrocytes.

[0072] The photosensitive gene is preferably ChR2. Viral vectors can be lentiviruses or adenoviruses.

[0073] Liposomes were used to transfect fusion plasmids containing light-sensitive genes, green fluorescent marker genes, and the promoter GFAP for specific labeling of glial cells, as well as mixed plasmids pCMVΔR8.74 and pMD2.G. related to lentiviral packaging, into 293FT cells , 6L liposomes per 200,000 293FT cells.

[0074] Among them, the structure of the fusion plasmid containing the photosensitive gene, the green fluorescent m...

Embodiment 1

[0106] Preparation of neural network repair system

[0107] (1) Construction of photosensitive astrocytes

[0108] The humanized GFAP-ChR2 codon (obtained from the laboratory of Professor Karl Deisseroth at Stanford University) was integrated into the N-terminal of the open framework plasmid pCS2-eYFP (purchased by Clotech) through the HindIII / BamHI restriction site. The successfully constructed plasmid was purified and amplified by Qiagen's maxiprep kit to obtain a fusion plasmid containing a light-sensitive gene, a green fluorescent marker gene, and a promoter GFAP for specifically labeling glial cells. The forward primer sequence for PCR amplification is the sequence shown in SEQ ID No.1, the reverse primer sequence is the sequence shown in SEQ ID No.2, and the source of GFAP is human.

[0109] Use liposomes to transfect 0.5 μg fusion plasmid containing photosensitive gene, green fluorescent marker gene and promoter GFAP for specific labeling of glial cells, and 1.5 μg mix...

Embodiment 2

[0148] Preparation of neural network repair system

[0149] (1) Construction of photosensitive astrocytes

[0150] Method is with embodiment 1.

[0151] (2) Detection of photosensitive astrocyte excitability and cell membrane integrity

[0152] Method is with embodiment 1.

[0153] (3) Construction of co-culture system of photosensitive astrocytes and stem cells

[0154] Method is with embodiment 1.

[0155] (4) Preparation of sustained release film

[0156] Method is with embodiment 1.

[0157] (5) Coat the slow-release film on the surface of the nerve electrode and dry it.

[0158] (6) Disperse the conductive polymer in the sustained release film

[0159] The nerve electrode coated with the hydrogel film is fully swollen in the electrodeposition solution containing the conductive polymer monomer, neurotrophic substance, nerve growth factor and ATP, so that the solution containing the conductive polymer monomer is evenly distributed on the swollen in the hydrogel film...

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Abstract

The invention relates to a nervous network restoration system and a preparation method thereof. The nervous network restoration system comprises a sustained-release diaphragm, a photosensitive astrocyte adsorbed on the sustained-release diaphragm, and a positively charged conducting polymer dispersed in the sustained-release diaphragm, wherein the sustained-release diaphragm is a hydrogel diaphragm formed by virtue of crosslinking of polyacrylic acid and polyvinyl alcohol. When the nervous network restoration system is used for treating an injured nervous network, the photosensitive astrocyte can specifically increase ATP (Adenosine Triphosphate) release under stimulation of light irradiation, negatively charged ATP is attracted by the positively charged conducting polymer to deposit on the surface of the sustained-release diaphragm, so that release of ATP is realized by diffusion, the defect of transient release of conventional drug treatment is overcome, stem cell proliferation and neuron directional differentiation can be promoted well, and the injured nervous network can be restored effectively.

Description

technical field [0001] The invention relates to the technical field of stem cell regulation, in particular to a neural network repair system and its preparation method and application. Background technique [0002] Stem cell replacement therapy has received more and more attention in the repair of damaged neural networks. Therefore, promoting the proliferation and directional differentiation of stem cells in organisms has always been the top priority in the field of stem cell research. Adenosine Triphosphate (ATP) is a nucleotide that acts as a "molecular currency" for intracellular energy transfer, storing and transferring chemical energy. As an important glial molecule, ATP participates in the regulation of other cells in the central nervous system, such as neurons and neural stem cells. A large number of reports have confirmed that ATP is involved in regulating the proliferation, migration and differentiation of stem cells (such as neural stem cells), thereby promoting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K50/00A61K9/70A61K41/00A61K47/32A61K35/30A61K31/7076A61P25/00A61P25/16A61P25/08A61P9/10A61P25/24A61P25/02A61L27/34A61L27/38A61L27/54C12N7/01C08J3/24C08L33/02C08L29/04G01N21/00G01N21/64
Inventor 屠洁鲁艺杨帆刘运辉王立平
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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