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Engineered nerve cell and preparation method and application thereof

A nerve cell and engineering technology, applied in the field of engineered nerve cells and their preparation, can solve the problems of long cycle, limited number of induction, complicated induction process, etc., and achieve the effects of short cycle, simple operation and uniform nature.

Pending Publication Date: 2021-03-19
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, iPSC and neuron cell culture and induction are difficult, the induction process is more complicated, and the cycle is long, and neuron cells can only differentiate but not proliferate, the number of each induction is limited, and it is difficult to meet the uniformity requirements for multiple inductions, which seriously affects AD. Research Reproducibility

Method used

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  • Engineered nerve cell and preparation method and application thereof
  • Engineered nerve cell and preparation method and application thereof
  • Engineered nerve cell and preparation method and application thereof

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Experimental program
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Embodiment 1

[0063] This embodiment prepares human-derived cells with Alzheimer's disease phenotype, and the preparation method of the human-derived cells with Alzheimer's disease phenotype includes the following steps:

[0064] (1) Culture human induced pluripotent stem cells (Cell Bank, Chinese Academy of Sciences, DYR0100) in 12-well plate with 1 mL of mTeSR1 medium containing 10 μM Rock inhibitor for 48 h;

[0065] (2) pTet-O-Ngn2-puro (purchased from addgene, Plasmid #52047), Tet-O-FUW-EGFP (purchased from addgene, Plasmid #30130) and FUdeltaGW-rtTA (purchased from addgene, Plasmid #19780) The plasmids were packaged into lentiviruses respectively, and the human-derived induced pluripotent stem cells were cultured for 6 hours using the mTeSR1 medium containing the lentiviruses, the medium containing the lentiviruses was aspirated, and replaced with fresh mTeSR1 medium for culture, and then replaced with fresh mTeSR1 every day Culture medium until the cells cover the bottom of the entir...

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Abstract

The invention provides an engineered nerve cell and a preparation method and application thereof. The engineered nerve cell is derived from engineered induced pluripotent stem cells, the engineered induced pluripotent stem cells are induced pluripotent stem cells containing plasmids, and the plasmids comprise pTet-O-Ngn2-puro, Tet-O-FUW-EGFP and FUdeltaGW-rtTA. According to the engineered nerve cell, the engineering induced pluripotent stem cells are rapidly prepared on a large scale through virus vector transduction, then the engineering nerve cells are obtained through antibiotic induction,screening and differentiation culture, and finally the beta-amyloid protein is used for inducing the engineering nerve cells to obtain the cells with Alzheimer's disease phenotypes. The whole preparation process is simple to operate and short in period, large-scale preparation can be achieved, and experimental materials which are uniform in property and repeatable are provided for drug developmentand research.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an engineered nerve cell and its preparation method and application. Background technique [0002] Alzheimer's disease (AD) is a neurodegenerative disease characterized by deposition of amyloid β-protein (Aβ), neurofibrillary tangles (NFTs) and senile plaques (SP) disease. At present, AD is an important research object in the field of neurological diseases, and an ideal AD model is an important basis for exploring its pathogenesis and developing new drugs. An important platform. Compared with in vivo models, in vitro models are more convenient and faster to obtain and establish, and their efficient operation and data acquisition are also very suitable for initial high-throughput drug screening. [0003] Aβ is a small molecule polypeptide processed by protease from amyloid precursor protein (APP), which can be divided into Aβ according to the amino acid sequence difference. 1-42 , Aβ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079C12N5/10C12N15/867
CPCC12N5/0618C12N15/86C12N2506/45C12N2740/15043C12N2501/998
Inventor 陈爽洪伟陈平
Owner SHENZHEN INST OF ADVANCED TECH