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Induction medium for inducing neural stem/progenitor cells to be differentiated into oligodendrocyte precursor cells and induction method and application thereof

A technology of induction medium and precursor cells, applied in the field of induction medium, can solve the problems of tumorigenesis, etc., and achieve the effect of broad application prospects, safe and effective treatment of myelin-related diseases

Active Publication Date: 2014-01-01
栾佐
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the totipotency of embryonic stem cells is a double-edged sword. It can not only differentiate into various cells, but also has the risk of tumorigenesis. Therefore, the OPCs obtained by this method still have a certain tumorigenic risk in clinical application.

Method used

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  • Induction medium for inducing neural stem/progenitor cells to be differentiated into oligodendrocyte precursor cells and induction method and application thereof
  • Induction medium for inducing neural stem/progenitor cells to be differentiated into oligodendrocyte precursor cells and induction method and application thereof
  • Induction medium for inducing neural stem/progenitor cells to be differentiated into oligodendrocyte precursor cells and induction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] In this example, OPCs were induced from the NSCs cell line established in our laboratory. The NSCs were derived from the hippocampus tissue of discarded embryos, and the NSCs were passaged to the 10th passage during induction.

[0050] 1. Digest NSCs into single cells, wash and resuspend in the pretreatment medium, and the cell suspension is 2×10 6 / T25 density seeded in T25 cell culture flasks, 8.5% CO 2 , 37°C, cultured under saturated humidity conditions.

[0051] 2. On the 4th day of cell culture, change the medium in half.

[0052] 3. On the 8th day of cell culture, change the medium in half.

[0053] 4. When the cells are cultured to the 12th day, collect the cells into a centrifuge tube and centrifuge at 400g for 5 minutes.

[0054] 5. The neural stem cell spheres were digested with 0.025% trypsin, the digestion was stopped with trypsin inhibitor, and the cells were pipetted into a single cell suspension.

[0055] 6. Collect the cells by centrifugation at 400...

Embodiment 2

[0063] Separate the brain tissue from the 10-week-old induced embryo brain tissue, mechanically disperse the tissue into a single-cell suspension, add pretreatment medium and culture, and the culture condition is 8.5% CO 2 , 37°C, saturated humidity, after the cells formed into spheres, they were passaged to induce OPCs.

[0064] The cultivation, induction and identification methods are the same as in Example 1. The identification results showed that the obtained OPCs highly expressed O4, NG2 and other OPCs markers, and the positive rate was 80-90%.

Embodiment 3

[0066] In this example, OPCs were induced from another NSCs cell line established in our laboratory. The NSCs were derived from the cortical tissue of discarded embryos, and the NSCs were passaged to the 25th passage during induction.

[0067] The cultivation, induction and identification methods are the same as in Example 1. The identification results showed that the obtained OPCs highly expressed O4, NG2 and other OPCs markers, and the positive rate was 80-90%.

[0068] The culture medium and composition that above-mentioned embodiment relates to are as follows:

[0069] DF medium formula:

[0070]

[0071] The pretreatment medium formula is as follows:

[0072]

[0073] OPCs induction medium formula is as follows:

[0074]

[0075]

[0076] DMEM and F12 are common media in cell culture, and can be purchased from any commercial company. In this implementation, these two media were purchased from Invitrogen, and the DMEM product number is 11965-118. For specifi...

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Abstract

The invention relates to the field of cytobiology and neurobiology, in particular to an induction medium for inducing neural stem / progenitor cells to be differentiated into oligodendrocyte precursor cells and an induction method and application thereof. The invention adopts the technical scheme that after subjected to preprocessing culture for a certain time by a preprocessing medium, the human neural stem / progenitor cells are rapidly differentiated into the oligodendrocyte precursor cells for expressing oligodendrocyte precursor cell markers such as O4, A2B5, NG2, SOX10, PDGFR (Platelet Derived Growth Factor Receptor) and the like at a high purity under the action of the induction medium; and the key ingredients of the induction medium are bFGF (Basic Fibroblast Growth Factor), PDGF-AA and NT-3 (neurotrophins-3). The method disclosed by the invention and the obtained oligodendrocyte precursor cells can be applied to preparation of a medicament for treating the nervous system injury disease and have wide prospect on the aspects of experiment research and clinical treatment.

Description

technical field [0001] The invention relates to the fields of cell biology and neurobiology, in particular to an induction medium for inducing neural stem / progenitor cells to differentiate into oligodendrocyte precursor cells, an induction method and application thereof. Background technique [0002] A variety of myelin-related diseases, such as brain white matter injury, spinal cord injury, multiple sclerosis and other diseases, due to myelinization disorder or demyelination of neurons, neurons can not normally transmit electrical excitation, resulting in the body Motor and other functional disabilities have brought great harm to patients and families of these diseases, but currently there is no drug that can effectively treat these myelin-related diseases. [0003] Whether it is a myelinating disorder or a demyelinating lesion, the root cause is the reduction or loss of the myelin sheath of the neuron axon. Therefore, if the degree of myelinization of the neuron axon can b...

Claims

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Application Information

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IPC IPC(8): C12N5/0797C12N5/079A61K35/30A61P25/00
Inventor 栾佐
Owner 栾佐
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