Subpopulation of oligodendrocyte precursor cells and uses thereof

A technology of precursor cells and cell subsets, applied in the field of biomedicine, can solve the problems of no specific drugs, decreased quality of life, and decreased survival rate.

Pending Publication Date: 2022-05-27
上海纽仁生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PSCI may lead to decreased quality of life, decreased survival rate, increased social burden, etc., and more and more attention has been drawn to the treatment of PSCI
[0004] At present, the tre...

Method used

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  • Subpopulation of oligodendrocyte precursor cells and uses thereof
  • Subpopulation of oligodendrocyte precursor cells and uses thereof
  • Subpopulation of oligodendrocyte precursor cells and uses thereof

Examples

Experimental program
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preparation example Construction

[0059] According to a specific embodiment of the present invention, the preparation method of the above exosome comprising:

[0060]1, the P6-P10 generation PDGFRA obtained in the method of preparing cell subsets proposed in the second aspect of the present invention, VCAN, LARS2, TTR, MAL, PCDH15 or CSPG4 positive oligodendrocytes are cultured with serum-free medium for more than 2 days, the cell supernatant is collected for exosome extraction; (The positive qualified refers to: the expression rate of the LARS2 protein is not less than 85%; the expression rate of the VCAN protein is not less than 90%; the expression rate of the VCAN protein is not less than 90%; The expression rate of the TTR protein is not less than 80%; the expression rate of the MAL protein is not less than 70%; the expression rate of the PCDH15 protein is not less than 80%; alternatively, the expression rate of the CSPG4 protein is not less than 80%).

[0061] 2. Centrifuge the supernatant of stable cell line...

Embodiment 1

[0097] Example 1 Single cell sequencing screening experiment identification

[0098] VCAN, LARS2, TTR, MAL, PCDH15, CSPG4 positive OPC cells

[0099] Single-cell transcriptome sequencing of brain tissue samples, the original data were obtained and the single-cell population was grouped according to the gene expression marker, and the OPC taxon was identified according to the expression of the Marker gene PDGFRA (see Annex for details Figure 1), using principal component analysis, cluster analysis, etc. to determine the expression specificity of different genes in different cells, and then screening markers that may be used for further grouping, the results showed that the grouping effect of VCAN, LARS2, TTR, MAL, PCDH15, and CSPG4 was better for subsequent further verification.

[0100] The inventor used iPSC to induce differentiation of cells positive for the above six genes (VCAN, LARS2, TTR, MAL, PCDH15, CSPG4), and verified the function of the cells, and found that the above c...

Embodiment 2

[0101] Example 2 iPSC induces differentiation of PDGFRA + VCAN + OPC cell preparation

[0102] Using purchased iPSC cells, incubate iPSC cells in a 6-well plate, culture media components (KnockoutDMEM, 1% non-essential amino acids, 2 mM L-glutamine, 100 Μ / mL penicillin, 100 μg / mL streptomycin, 100-LM 2-mercaptoethanol), culture for 7 days, and then exchange neural precursor cells (NPC) for differentiation of serum-free N2 medium medium (DMEM / F12, 2% N2 supplement, 2 mM L-glutamine, Penicillin / streptomycin, 1% sodium pyruvate + 20 ng / mL alkaline fibroblast growth factor (FGF2) and 20 ng / mL epidermal growth factor (EGF) for 7 days; to induce differentiation into OPC cells, neuroprotec cells (NPCs) are inoculated on a patch coated with poly-D-lysine (PDL, 10 lg / mL)-laminin (5 lg / mL) and 10 ng / mL platelet-derived growth factor (PDGF) is added to the cap coated with poly-D-lysine (PDL) - laminin (5 lg / mL) Continue in culture in N2 medium for 4 days, the previous 2 days N2 medium also c...

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Abstract

The invention provides an oligodendrocyte precursor cell subset, which is characterized in that the oligodendrocyte precursor cell subset is positive for at least one of the following markers: VCAN, LARS2, TTR, MAL, PCDH15 and CSPG4. The cell subset provided by the invention is easy to screen and prepare, and plays an important role in preparing medicines, especially medicines for treating or preventing nerve, blood vessel or muscle injury diseases.

Description

Technical field [0001] The present invention belongs to the biomedical field, involving oligodendrocyte precursor cell subsets, specifically, exosomes involving oligodendrocyte precursor cells subpopulation, oligodendrocyte precursor cell subset preparation method, oligodendrocyte precursor cell subset and its exosomes in the preparation of drugs, methods for screening drugs, pharmaceutical compositions. Background [0002] Oligodendrocyte Precursor Cells (OPC) are a group of cells with stem cell characteristics that are localized in the nervous system with high proliferation and differentiation ability, OPC cells can differentiate into oligodendrocytes and may be involved in diseases caused by oligodendrocyte abnormalities. Exosomes (exosomes) are tiny membrane vesicles that can be secreted by most cells in the body, with a lipid bilayer membrane, about 30 to 150 nm in diameter. Exosomes are widely present and distributed in various body fluids, carrying and transmitting importa...

Claims

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Application Information

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IPC IPC(8): C12N5/079C12Q1/02A61K35/30A61P25/28A61P9/10
CPCC12N5/0622G01N33/5058G01N33/5038A61K35/30A61P25/28A61P9/10C12N2503/02G01N2500/00
Inventor 张允斌
Owner 上海纽仁生物医药科技有限公司
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