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Human neural stem cell culture medium and application

A culture medium and stem cell technology, applied in the field of cell culture, can solve the problems of severe spontaneous differentiation and limited proliferation ability of human neural stem cells, and achieve the effect of easy differentiation and difficult long-term expansion, low cost and simple formula

Active Publication Date: 2018-02-06
BEIJING TRANSGEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the proliferation ability of human neural stem cells is limited in the existing medium, and the spontaneous differentiation is relatively serious.

Method used

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  • Human neural stem cell culture medium and application
  • Human neural stem cell culture medium and application
  • Human neural stem cell culture medium and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Human neural stem cell culture medium

[0033] A culture medium for human neural stem cells, said medium comprising the following components:

[0034] Basal medium (volume ratio): 50% DMEM / F12 medium and 50% Neurobasal medium.

[0035] Nutritional additives: the components and final use concentrations of the nutritional additives are: human insulin 10mg / L, vitamin C 50mg / L, glutathione 50mg / L, linolenic acid 1mg / L, carnitine 20mg / L, N-acetylcysteine ​​5μM, ethanolamine 5mg / L, linoleic acid 1mg / L.

[0036] Growth factor components and their final use concentration: bFGF: 20ng / mL; EGF: 20ng / mL.

[0037]The signaling pathway regulates small molecule components and their final use concentrations: Pifithrin-αhydrobromide, 5μM and 6-BIO, 2μM.

Embodiment 2

[0038] Example 2 Culture medium for human neural stem cells

[0039] A culture medium for human neural stem cells, said medium comprising the following components:

[0040] Basal medium (volume ratio): 50% DMEM / F12 medium and 50% Neurobasal medium.

[0041] Nutritional additives: the composition and final use concentration of the nutritional additives are: human insulin 0.1mg / L, vitamin C 10mg / L, glutathione 10mg / L, linolenic acid 0.05mg / L, carnitine 0.2mg / L, N-acetylcysteine ​​5 μM, ethanolamine 0.01 mg / L, linoleic acid 0.05 mg / L.

[0042] Components of growth factors and their final use concentrations: bFGF: 5ng / mL; EGF: 5ng / mL.

[0043] The signaling pathway regulates small molecule components and their final use concentrations: WNT3a, 1ng / mL and Baxinhibitor peptide V5, 0.1μM and Cyclic Pifithrin-αhydrobromide, 0.1μM.

Embodiment 3

[0044] Example 3 Culture medium for human neural stem cells

[0045] A culture medium for human neural stem cells, said medium comprising the following components:

[0046] Basal medium (volume ratio): 50% DMEM / F12 medium and 50% Neurobasal medium.

[0047] Nutritional additives: the components and final use concentrations of the nutritional additives are: human insulin 20mg / L, vitamin C 200mg / L, glutathione 100mg / L, linolenic acid 5mg / L, carnitine 20mg / L, N-acetylcysteine ​​500μM, ethanolamine 10mg / L, linoleic acid 5mg / L.

[0048] Components of growth factors and their final use concentrations: bFGF: 100ng / mL; EGF: 100ng / mL.

[0049] The signaling pathway regulates small molecule components and their final use concentrations: WNT3a, 1ng / mL and Baxinhibitor peptide V5, 100μM and Cyclic Pifithrin-αhydrobromide, 100μM.

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Abstract

The invention discloses a human neural stem cell culture medium. The culture medium is prepared from the following components: a basic culture medium, a nutrient additive, a growth factor and a signalpathway regulation and control micro-molecule. The invention further discloses application of the culture medium to culturing of human neural stem cells. The culture medium disclosed by the inventionhas the advantages of simple formula, low cost and clear chemical components; the culture medium does not contain animal-derived components and does not contain protein extract and hydrolysate; the culture medium of all batches is uniform and stable and has no potential risks of introducing animal-derived and human-derived pathogenic microorganisms, so that the culture medium is safer and can beapplied to the culturing of the human neural stem cells in clinical researches and clinical experiments; furthermore, the signal pathway regulation and control micro-molecule is applied to the culturing of the human neural stem cells for the first time; under the action of the signal pathway regulation and control micro-molecule, rapid proliferation and dryness keeping of the human neural stem cells with various sources can be supported for a long period; the technical problem that the human neural stem cells are easy to differentiate and are difficult to proliferate for a long period when thehuman neural stem cells are cultured in vitro is successfully solved.

Description

technical field [0001] The invention relates to the technical field of cell culture. More specifically, it relates to a human neural stem cell culture medium and its application. Background technique [0002] The incidence of various nerve injuries and degenerative neurological diseases is increasing year by year. Traditional medical methods are difficult to cure these diseases completely. Cell replacement therapy is the most effective means to cure these diseases. [0003] Neural stem cells have the ability to self-replicate and differentiate into cell types such as neural precursor cells, neurons, astrocytes, and oligodendrocytes. They are important tools for disease pathogenesis research, drug screening, and neurodegenerative diseases. It is an ideal seed resource cell for cell replacement therapy of damaged and damaged diseases. At present, there are many ways to obtain neural stem cells in vitro, such as primary isolation and culture from brain tissue, induction and d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797
CPCC12N5/0623C12N2501/10C12N2501/11C12N2501/115C12N2501/40C12N2501/415C12N2501/48
Inventor 王娟马静辛文
Owner BEIJING TRANSGEN BIOTECH CO LTD
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