Method for promoting differentiation of rat radial glial cells into neurons

A glial cell and radial technology, applied in the field of promoting the differentiation of radial glial cells into neurons in rats, can solve the problems of neuron degeneration and necrosis

Pending Publication Date: 2020-10-30
NANTONG UNIVERSITY
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Problems solved by technology

At present, the main method for clinical treatment of such diseases is to supplement the missing neurotransmitters, but this cannot fundamentally solve the problem of neuron degeneration and necrosis

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  • Method for promoting differentiation of rat radial glial cells into neurons
  • Method for promoting differentiation of rat radial glial cells into neurons

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Experimental program
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Embodiment 1

[0074] 1. Isolation and culture of rat radial glial cells:

[0075] (1) Take one 14-day-pregnant SD rat, anesthetize it intraperitoneally with compound anesthetic Chlorpent 0.4ml / 100g, open the abdominal cavity under aseptic conditions, and place it in a petri dish containing pre-cooled DMEM / F12 basal culture medium;

[0076] (2) Take out the whole brain tissue of the fetal mouse on the ultra-clean bench, and cut the cortex into pieces of 1mm 3 the size of the tissue block;

[0077] (3) Move the cortical tissue pieces to a centrifuge tube filled with an appropriate amount of basic culture medium, mechanically blow gently until no obvious tissue pieces, filter with a sterile 200-mesh nylon mesh, and collect the filtered cell suspension;

[0078] (4) Resuspend the cells with RGCs culture medium, stain and count with trypan blue, count as 1×10 5 Inoculate at a density of RGCs / ml in a cell culture flask containing 10ml RGCs culture solution, place at 37°C, 5% CO 2 incubator cul...

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Abstract

The invention discloses a method for promoting differentiation of rat radial glial cells into neurons, comprising the following steps of: acquiring a fetal rat cerebral cortex cell suspension, and preparing RGCs single cells; and inoculating the RGCs single cells into a culture plate, culturing overnight by using an RGCs culture solution, and continuously culturing by using an RGCs induced differentiation solution. The method for preparing the RGCs single cells comprises the following steps of: resuspending cells by using an RGCs culture solution to form macroscopic suspended cell spheres, replacing the RGCs culture solution, and continuously culturing for 5-7 days to obtain the cell spheres; and treating the cell spheres with Accutase enzyme, digesting the cell spheres into single cells,continuing suspension culture, and carrying out passage. According to the method, the differentiation ratio of the rat embryo cerebral cortex derived radial glial cells to neurons can be obviously improved and can reach about 3 times or even higher than the natural differentiation rate.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a method for promoting the differentiation of rat radial glial cells into neurons. Background technique [0002] The important pathological basis of neurodegenerative diseases is the degeneration and necrosis of neurons in local areas, resulting in the destruction of neural circuits or the loss of neurotransmitters. At present, the main method for clinical treatment of such diseases is to supplement the missing neurotransmitters, but this cannot fundamentally solve the problem of neuron degeneration and necrosis. The use of neural stem cells (Neural stem cells, NSCs) with multi-directional differentiation potential for replacement therapy is considered to be an ideal method, but under natural differentiation conditions in vivo and in vitro, the number of NSCs differentiated into neurons is very small, which is There is a need to guide NSCs to differentiate into ...

Claims

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Application Information

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IPC IPC(8): C12N5/079C12N5/0793
CPCC12N5/0622C12N5/0619C12N2509/00C12N2501/11C12N2501/115C12N2501/13
Inventor 张蕾张新化赵荷艳
Owner NANTONG UNIVERSITY
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