Neural stem cell culture medium and culture method

A technology of neural stem cells and nerve cells, applied in nervous system cells, cell culture active agents, animal cells, etc., can solve problems affecting the application and research of NSCs nervous system damage diseases, avoid moral disputes, and reduce non-immune rejection Response, the effect of convenient material collection

Active Publication Date: 2017-03-08
SHANGHAI AISAER BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In summary, the source of NSCs is often limited by existing technologies or ethics, which ...

Method used

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  • Neural stem cell culture medium and culture method
  • Neural stem cell culture medium and culture method
  • Neural stem cell culture medium and culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1~4

[0035] Examples 1-4 use peripheral blood mononuclear cells to cultivate neural stem cells

[0036] 1. Culture medium

[0037] The culture solutions of Examples 1-4 include a basal medium formed by mixing DMEM / F12 medium and neurobasal medium, and additives added to the basal medium. Table 1 shows the mixing volume ratio, additives and contents of DMEM / F12 medium and nerve cell basal medium in each embodiment.

[0038] Table 1 The composition and content of the culture medium

[0039]

[0040] Remarks: All substances mentioned in Table 1 are purchased from the market

[0041] 2. Training process

[0042] Examples 1-4 respectively used the corresponding culture medium shown in Table 1 to culture neural stem cells according to the following steps.

[0043] 1. Separation of peripheral blood mononuclear cells by Ficoll-Paque density gradient centrifugation

[0044] by SepMate TM Add 3.5mL Ficoll-Paque (relative density 1.077) into the small hole in the middle of the tube. ...

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Abstract

The invention discloses a culture medium for culturing neural stem cells on the basis of peripheral blood mononuclear cells. The culture medium comprises components as follows: a DMEM/F12 culture medium and a neurobasal medium in a volume ratio being 1:0.8-1.2 as well as an adenylate cyclase activator, a serum substitute, L-ascorbic acid, an L-glutamine derivative, an ALK inhibitor, a GSK-3 inhibitor, an ALK-2,3,6,AMPK inhibitor and an ALK-4,5,7 inhibitor. The invention further discloses a method for culturing the neural stem cells on the basis of the peripheral blood mononuclear cells. With adoption of the culture medium, the neural stem cells can be obtained from the peripheral blood mononuclear cells as the raw material through directional differentiation culture, materials are taken conveniently and free of quantitative restriction, and moral controversy can be avoided.

Description

technical field [0001] The invention belongs to the technical field of neural stem cell culture, and in particular relates to a culture medium and a culture method of neural stem cells. Background technique [0002] Neural stem cells (NSCs) are a kind of mother cells with division potential and self-renewal ability, which can produce various types of cells in nervous tissue through asymmetric division, including neurons, oligodendrocytes and astrocytes. The use of transdifferentiation technology can achieve NSCs proliferation, self-renewal and multidirectional differentiation in vitro. However, a small amount of NSCs isolated from the human body (such as the central nervous system) can be differentiated into corresponding NSCs under the influence of the microenvironment in the body after being transplanted into the human body through proliferation to a certain concentration and standard, so that the damaged nerve function can be repaired. , and the implanted NSCs can migrat...

Claims

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Application Information

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IPC IPC(8): C12N5/0797
CPCC12N5/0623C12N2500/30C12N2500/38C12N2501/998C12N2506/115
Inventor 高歌陈瑛
Owner SHANGHAI AISAER BIOTECH CO LTD
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