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Method for promoting symmetrical division of neural stem cells

A neural stem cell, symmetrical technology, applied in the biological field, can solve problems such as the difficulty of obtaining neural stem cell research, the slow proliferation of neural stem cells, and the difficulty of harvesting neural stem cells, so as to save working time, save time and energy, and the preparation method is simple Effect

Inactive Publication Date: 2017-09-26
温州市鹿城区中津先进科技研究院
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Problems solved by technology

In this suspension culture method, neural stem cells proliferate slowly, and their viability is very poor, and they are very prone to apoptosis, so it is difficult to harvest a large number of neural stem cells
Therefore, this makes it difficult for relevant scientific researchers to obtain a large number of neural stem cells for related research work
In order to solve the problem of traditional in vitro culture of neural stem cells, neurospheres are cultured in suspension. After the number of neurospheres is doubled, centrifugation and bottle expansion are used to achieve the purpose of subculture. In this method of suspension culture, neural stem cells proliferate slowly and have poor viability. Very easy to apoptosis, difficult to harvest a large number of neural stem cells and other issues

Method used

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Embodiment Construction

[0019] Below by specific embodiment the present invention is described in further detail.

[0020] The methods used in the embodiments of the present invention include nerve tissue preservation solution, nerve tissue washing solution, nerve tissue decomposition solution, cell washing solution, primary cell culture solution, cell digestion solution, subculture cell culture solution, cell cryopreservation solution, etc. Wherein, the secondary cell culture medium is a serum-containing growth medium for neural stem cells, which consists of 1000ml basal medium, 50ug / ml fetal bovine serum, 10ng / ml bFGF, 10ng / ml EGF, 0.5mmol / ml ml glutamine, 0.5mmol / ml sodium pyruvate mixed composition. The secondary cell culture medium is the obtained neural stem cell adherence culture medium, which enables the neural stem cells to adhere to the wall and have stronger activity. The nerve tissue preservation solution is a culture medium containing one or more of RPMI-1640, DMEM, a-MEM and DMEM / F12 c...

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Abstract

The present invention provides a method for promoting symmetrical division of neural stem cells, which can be supplemented by selecting cells expressing neural stem cell-specific markers, for example linking such expression markers to their expression in differentiated progeny, preferably in neural stem cell active promoter. The method preferably comprises passage cells at or below 65% confluence, more preferably at or below 55% confluence, especially below 50% confluence.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for promoting the symmetrical division of neural stem cells. Background technique [0002] Neural stem cells are a kind of mother cells with division potential and self-renewal ability, which can produce various cells of nerve tissue through asymmetric division. According to the timing of differentiation potential, it can be divided into: neural tube epithelial cells, radial glial neurons and neuroblasts mainly in the embryonic period. Among them, radial glial neurons mainly exist in the juvenile period, which can divide and produce neuron precursors at the same time Cells or glial cells; neuroblasts mainly exist in adult animals and can produce neural precursor cells, neurons and various types of glial cells. According to the location, there are mainly two types: neural crest stem cells and central nervous stem cells. At present, the cells that have the ability to differe...

Claims

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Application Information

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IPC IPC(8): C12N5/0797
CPCC12N5/0623C12N2500/30C12N2500/32C12N2501/11C12N2501/115
Inventor 王振宇
Owner 温州市鹿城区中津先进科技研究院
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