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Method of promoting differentiation of rat neural stem cells

A neural stem cell and neuron technology, applied in the field of promoting rat neural stem cell differentiation, can solve the problems of high cost of complete culture medium and low differentiation yield, and achieve the effects of simple components, low cost and increased quantity

Inactive Publication Date: 2013-10-09
天津市环湖医院
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Problems solved by technology

[0010] The present invention aims to solve the problem of high cost and low differentiation yield of the existing neuron complete culture medium in creating a microenvironment for the differentiation of stem cells at the transplantation site, and provides a neural stem cell-directed neuron with simple components and low cost. Method for promoting the differentiation of rat neural stem cells for neuron differentiation

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  • Method of promoting differentiation of rat neural stem cells
  • Method of promoting differentiation of rat neural stem cells
  • Method of promoting differentiation of rat neural stem cells

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Embodiment Construction

[0026] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0027] In order to obtain neuronal cells with higher purity, neural stem cells must be cultured first, and then differentiated and then identified.

[0028] 1. Fetal mouse brain neural stem cell culture, the specific steps are as follows:

[0029] 1. SD rats on the 13th day of pregnancy were killed by dislocation of the neck. After being sterilized with 0.1‰ brogeramine, the embryos were taken by laparotomy, the placenta and skull were removed, the brain tissue was taken, washed 3 times with culture medium, and shredded fully;

[0030] 2. Digest into single cells with 0.25% trypsin and 0.02% EDTA disodium, centrifuge at 1500r / min for 10 minutes;

[0031] 3. Suspend the cells with the complete culture medium of neural stem cells, pass through a 200-mesh sieve, collect the cell filtrate, and place it at 75 cm 2 In a culture flask, at 37°C, with a volu...

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Abstract

The invention discloses a method of promoting differentiation of rat neural stem cells, belonging to culture of undifferentiated cells. The method comprises shredding brain tissues of fetal rats in 13-day pregnant SD rats, digesting into unicells, centrifuging, suspending the cells with a neural stem cell complete culture medium, sieving, culturing to obtain high-purity brain neural stem cells; collecting and centrifuging neural stem cells of a second generation, inoculating the cells with a neuron complete medium together with a 5 % fetal calf serum onto a 24-well plate already coated with rat tail collagen, adhering the cells onto the wall, starting to use the neuron complete medium at the first day of cell differentiation, simultaneously adding 10 [mu]l of 100 * 100 [mu]mol / L taurine into each hole, and culturing for 12 days to obtain the neural stem cells with differentiated neurons. Beneficial effects of the invention are that components of the nutrient solution are simple, cost is low, the added taurine exists in the body and thus usage is safe, and an amount of the generated mature neurons can be increased.

Description

technical field [0001] The invention relates to the cultivation of undifferentiated cells, in particular to a method for promoting the differentiation of neural stem cells into neurons in rats. technical background [0002] Stem cells are primitive cells with the potential of self-replication and multi-directional differentiation. They are the origin cells of the body and the primitive cells that form various tissues and organs of the human body. Neural stem cells have the potential to differentiate into neurons, astrocytes and oligodendrocytes. The main characteristics of neural stem cell differentiation are: ①the plasticity of neural stem cells; ②directed differentiation of neural stem cells: including the differentiation of neural stem cells into neurons and glia; ③transverse differentiation of neural stem cells; ④induced differentiation of neural stem cells; ⑤implantation Differentiation of neural stem cells in vivo. [0003] Neural stem cells have brought new hope to ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0793
Inventor 黄慧玲范维佳武俏丽蔡英王辰莫丽冬
Owner 天津市环湖医院
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