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Clinical-grade neural stem cell culture medium and preparation method thereof

A neural stem cell and culture medium technology, applied in the fields of neurobiology and cell biology, can solve the problem of poor stem cell culture effect, complex medium composition, cell stability, stemness retention effect and expansion rate to be further improved, etc. problems, to achieve high promotion and application value, reduce the amount of serum added, and maintain good dryness.

Pending Publication Date: 2021-04-09
CHENGDU ZHIA TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the composition of the medium is complex, the effect of stem cell culture is not good, and the cell stability, stemness maintenance effect and expansion rate need to be further improved

Method used

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  • Clinical-grade neural stem cell culture medium and preparation method thereof

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Effect test

Embodiment 1

[0025]A medium suitable for large-scale culturing clinical neural stem cells, characterized in that the components are as follows: base medium 13g / L, insulin growth factor-1 1 ng / ml, transforming growth factor β1 1 ng / ml, epidermal growth factor 3 ng / ml, fetal bovine serum 5 ng / ml, sodium pyruvate 0.001 mg / ml, glutamine 0.01 mm, chloromethythyltrimetarne modified adenosine 120 μm, composite vitamin 5 μm, hyperbranched polyamino acid 3 μm, super branched The polysaccharide was 1 μm, 3 μg / ml of carbooquin, 0.008 mg / ml of sodium chloride, 0.008 mg / ml of sodium hydrogencarbonate.

[0026]The base medium is DMEM; the medium suitable for large-scale culturing clinical neural stem cells further includes 4 nm of female glycol.

[0027]The preparation method of the chloromethythyltrimethyltrimetarne alkane-modified adenosine includes the step of: adding chloromethyltrimethyltrimetarne, adenosine to an organic solvent, stirred at 40 ° C for 6 hours, and then Decanogenate, washed 3...

Embodiment 2

[0031]A medium suitable for large-scale culturing clinical neural stem cells, characterized in that the components are as follows: base medium 15g / L, insulin growth factor-1 2.5 ng / ml, transforming growth factor β1 1.5 ng / ml, epidermal Growth factor 5 ng / ml, fetal bovine serum 7 ng / ml, pyruvate 0.0015 mg / ml, glutamine 0.015 mm, chloromethythyltrimetarne modified adenosine 140 μm, composite vitamin 20 μm, super branched polyamino acid 8 μm, Super branched polysaccharide 3 μm, 4 μg / ml of carbinianilin, 0.009 mg / ml of sodium chloride, 0.009 mg / ml of sodium bicarbonate.

[0032]The base medium is an IMDM medium; the medium suitable for large-scale culture clinical neural stem cells also includes: pentyl glycol 6 nm.

[0033]The method of modulating adenosine of chloromethyltrimethyltrimide, comprising the step of: adding chloromethythyltrimethyltrimetarne, adenosine to an organic solvent, stirred at 45 ° C for 6.5 hours, and then The solvent was evaporated, washed 4 times wi...

Embodiment 3

[0037]A medium suitable for large-scale culturing clinical neural stem cells, characterized in that the components are as follows: base medium 16g / L, insulin growth factor-1 3.5 ng / ml, transforming growth factor β1 2 ng / ml, epidermal growth Factor 7 ng / ml, fetal bovine serum 8 ng / ml, pyruvate 0.002 mg / ml, glutamine 0.02 mm, chloromethyltrimethyride modified adenosine 190 μm, compound vitamin 35 μm, hyperbranched polyamino acid 12 μm, super branch 5 μm of polysaccharide, 4.5 μg / ml of carbooquelin, 0.001 mg / ml of sodium chloride, 0.01 mg / ml of sodium bicarbonate.

[0038]The base medium is an ES medium; the medium suitable for large-scale culturing clinical neural stem cells also includes 8 nm of pentyl glycol.

[0039]The preparation method of the chloromethyltrimethyltrimethyl glycoline modification adenosine includes the step of: adding chloromethyltrimetarne nutane, adenosine to an organic solvent, stirred at 50 ° C for 7 hours, and then The solvent was distilled, the ...

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Abstract

The invention discloses a culture medium suitable for large-scale culture of clinical-grade neural stem cells. The culture medium is characterized by being prepared from the following components: 13 to 20g / L of a basic culture medium, 1 to 5ng / mL of an insulin growth factor-1, 1 to 3ng / mL of a transforming growth factor beta 1, 3 to 10ng / mL of an epidermal growth factor, 5 to 12ng / mL of fetal calf serum, 0.001 to 0.003mg / mL of sodium pyruvate, 0.01 to 0.03mM of glutamine, 120 to 220 microms of chloromethyl trimethyl germane modified adenosine, 5 to 50 microms of compound vitamin, 3 to 15 microms of hyperbranched polyamino acid, 1 to 7 microms of hyperbranched chitosan, 3 to 6 micrograms / mL of carbenicillin, 0.008 to 0.012mg / mL of sodium chloride and 0.008 to 0.012mg / mL of sodium bicarbonate. The invention further provides a preparation method of the culture medium suitable for the large-scale culture of the clinical-grade neural stem cells. The culture medium suitable for the large-scale culture of the clinical-grade neural stem cells, disclosed by the invention, has the advantages of high safety, good stem cell culture effect, good cell stability and stem property keeping effect and rapid amplification speed.

Description

Technical field[0001]The present invention relates to the field of cell biology, neurological biological technology, and more particularly to a medium suitable for mass culture clinical neural stem cells and a preparation method thereof.Background technique[0002]The central nervous system injury was considered to be unpaid, traditional drugs and other treatments were also maintained by maintaining existing neuronal survival and function. In recent years, with the research and development of stem cell technology, the researchers have found that neurons lost in injury during the occurrence of nervous system damage, can be recovered by transplanting stem cells, and this method is very effective. It is in this situation, and the in vitro of neural stem cells is cultivated into one of the popular topics of the current industry researchers.[0003]Neural stem cells refers to a cell population in an earlier developmental stage in the nervous system, which can be self-replicated by themselves...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797
CPCC12N5/0623C12N2500/30C12N2500/32C12N2500/34C12N2500/38C12N2500/40C12N2501/105C12N2501/11C12N2501/15C12N2501/392
Inventor 向兴力
Owner CHENGDU ZHIA TECH CO LTD
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