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Human neural stem cells expressing human choline acetyltransferase, and use thereof

A choline acetyl and stem cell technology, applied in the field of human neural stem cells and their uses, can solve problems such as tissue damage

Active Publication Date: 2013-07-03
DESIGNED CELLS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the drug treatment without a strong protective effect on tissue damage is only a temporary symptom improvement effect. The stem cell therapy method for targeted therapy of Alzheimer's disease can meet the above requirements

Method used

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  • Human neural stem cells expressing human choline acetyltransferase, and use thereof
  • Human neural stem cells expressing human choline acetyltransferase, and use thereof
  • Human neural stem cells expressing human choline acetyltransferase, and use thereof

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Embodiment approach

[0060] Hereinafter, the present invention will be described in more detail through examples. For those skilled in the art of the present invention, these embodiments are only provided to illustrate the present invention more specifically. According to the gist of the present invention, it is obvious that the scope of the present invention is not limited to these embodiments.

[0061] 【Example】

Embodiment 1

[0062] [Example 1: Human choline acetyltransferase (ChAT) cloning and cDNA construction]

[0063] The full-length cDNA of human choline acetyltransferase (ChAT, choline acetyltrnasferase) was obtained from human small intestine Marathon-Ready cDNA (yeast double Hybrid Systems Inc., Mountain View, CA; Clontech, Mountain View, CA), obtained by the polymerase chain reaction (PCR) method. Sequence analysis of plasmid DNA was performed using an ABI3100 DNA Sequencer (Applied Biosystems, Foster City, CA: Applied Biosystems, Foster City, CA).

Embodiment 2

[0064] [Example 2: Human neural stem cells (Neural Stem Cells, NSCs)]

[0065] Infect the HB1.F3 (F3) human neural stem cell (NSC, neural stem cell) cell line with the retrovirus containing the pLPCX.ChAT gene containing the full-length cDNA of human choline acetyltransferase (8, 9, 11, 28 ), and screen for puromycin-resistant cells (13). F3.ChAT of choline acetyltransferase overexpressing clones were isolated, and the expression of choline acetyltransferase was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence microscopy.

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Abstract

The present invention relates to human neural stem cells which are transformed with a vector comprising a nucleotide sequence encoding human choline acetyltransferase (ChAT) to express the ChAT enzyme, a pharmaceutical composition for treating Alzheimer's disease, containing the same, and a composition for improving cognitive disorders that affect learning or memory ability. If transplanted to a brain of an animal model with Alzheimer's disease, the human neural stem cells genetically modified to express human ChAT were successfully combined to the brain tissue of a host to be differentiated into normal nerve cells, and stably expressed ChAT, thereby replenishing the reduced concentration of acetylcholine to a normal level. In addition, the human neural stem cells improve the cognitive function of an animal model with Alzheimer's disease in which cognitive functions are damaged. Therefore, the human neural stem cells expressing ChAT of the present invention can be developed as a drug for treating Alzheimer's disease and improving cognitive disorders that affect learning or memory ability caused by the disease.

Description

【Technical field】 [0001] The present invention relates to a kind of human neural stem cell and its application. The human neural stem cell is transformed into a vector containing the nucleotide sequence encoding human choline acetyltransferase to express choline acetyltransferase. In more detail, the present invention relates to Alzheimer's A pharmaceutical composition for treating Haimer's disease and a composition for improving cognitive dysfunction, the composition comprising human neural stem cells expressing the above-mentioned human choline acetyltransferase. 【Background technique】 [0002] Alzheimer's disease (AD) is one of the most devastating neurodegenerative diseases, characterized by impairment of cognitive functions such as learning and memory impairment. (1) Reduced activity of choline acetyl transferase (ChAT), which is an acetylcholine (ACh) synthetase, was observed in patients with Alzheimer's disease, while the activity of the presynaptic cholinergic system...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797C12N5/10C12N15/54C12N15/85
CPCC12N5/0623C12N9/1029C12Y203/01006C12N2510/00A61K48/005A61P25/28A61K35/30
Inventor 金允培金承业
Owner DESIGNED CELLS CO LTD