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Application of endothelial cells and precursor cells thereof in treatment of demyelination diseases

A demyelinating disease and precursor cell technology, applied to the application field of endothelial cells and their precursor cells in the treatment of demyelinating diseases, can solve problems such as large side effects and ineffective myelin repair

Pending Publication Date: 2021-12-28
呈诺再生医学科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most of the drugs used to treat demyelinating disease models are immunomodulators, such as glucocorticoids: prednisone, dexamethasone, methylprednisolone; immunoglobulin and interferon β-1, but the above drugs have long-term The use of side effects is large, such as obesity, insulin resistance, abnormal blood lipid and blood pressure, peptic ulcer, osteoporosis complicated by osteonecrosis, rash, acute renal failure, etc., and these drugs can slow down the disease process, inhibit inflammation, and play a certain role in treatment effect, but not on myelin repair

Method used

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  • Application of endothelial cells and precursor cells thereof in treatment of demyelination diseases
  • Application of endothelial cells and precursor cells thereof in treatment of demyelination diseases
  • Application of endothelial cells and precursor cells thereof in treatment of demyelination diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Example 1 Effect of hiPSC-EC transplantation on myelination

[0099] 1. Establishing a mouse model of nerve demyelination

[0100] 1% lysoletinin (Lyso) was directly injected into the mouse corpus callosum (CC) site to cause nerve demyelination (injection model diagram see figure 1 A), specific method references (A new EAE model of braindemyelination induced by intracerebroventricular pertussis toxin. Biochem Biophys Res Commun. 2008 May 23; 370(1): 16-21). Simultaneous use of cyclosporine to prevent xenogeneic rejection.

[0101] 2. Induced differentiation from iPSCs to form EPC / EC cells

[0102] Steps: using patented technology (see publication number: CN108384746B, patent name: a method for efficient differentiation of induced pluripotent stem cells into mature endothelial cells), induce differentiation of iPSCs to form EPC / EC cells.

[0103] 3. Detection of BDNF secreted by EPCs induced by iPSCs

[0104] Steps: EPCs induced by iPSCs were fixed with 4% paraformal...

Embodiment 2

[0111] Example 2 Effect of hiPSC-EC transplantation on nerve remyelination and axon repair

[0112] To evaluate the effect of hiPSC-EC (EC derived from human induced pluripotent stem cells) transplantation on neuronal protection against demyelination-induced axonal injury, axons from the demyelinated area of ​​corpus callosum (CC) were treated with NF200 mark. hiPSC-EC transplantation effectively increased the staining intensity of NF200 markers (using FujiIntDen to count the fluorescence intensity, 0.82 in the hiPSC-EC group was higher than 0.24 in the control group, p<0.05), and the NF200+ fiber structure was clearer, implying neuroprotection or rapid recovery . This axonal protection is associated with myelin lipid protection / remyelination promoted by iPSC-EC transplantation. Note: The control group was injected with the same volume of vehicle (i.e. PBS solution) as the hiPSC-EC group in the same way.

Embodiment 3

[0113] Example 3 Effect of hiPSC-EC transplantation on recruiting OPC enrichment

[0114] 1. Steps

[0115] 1% lysoletinin (Lyso) was directly injected into the mouse corpus callosum (CC) site to cause nerve demyelination, the specific method reference (A new EAE model of brain demyelination induced by intracerebral pertussis toxin.Biochem Biophys Res Commun.2008May23; 370 (1):16-21). Simultaneous use of cyclosporine to prevent xenogeneic rejection. After 3 days of modeling, hiPSC-ECs were transplanted in the injured area, and the enrichment of OPCs in the injured area was observed 8 days later. In order to better observe the enrichment of endogenous OPC, the above experiment was carried out using a mouse strain carrying the SOX10-GFP reporter gene (a gift from Professor W. Richardson of UCL University, UK).

[0116] 2. Results

[0117] The results showed that OPC enrichment could be seen in the injured area after transplantation ( image 3 ).

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Abstract

The invention discloses an application of endothelial cells and precursor cells thereof in treatment of demyelination diseases. The research finds that the endothelial cells and the precursor cells thereof can promote OPC cell proliferation, migration and maturation, promote astrocyte reaction and promote microglial cell / macrophage activation so as to promote myelin sheath formation, regeneration and repair. The research result of the invention provides a method for actively regulating and controlling myelin sheath formation and regeneration for clinic.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to the application of endothelial cells and their precursor cells in the treatment of demyelinating diseases. Background technique [0002] Myelin is the fatty, white substance that surrounds the axons of certain nerve cells, forming an electrically insulating layer. In humans, approximately 40% of the brain contains white matter comprising densely packed fibers of which myelin is the major component (50-60% dry weight of white matter). Myelin is synthesized and maintained by oligodendrocytes in the central nervous system (CNS). Oligodendrocytes are a type of glial cell that function to provide support and insulation for axons in the CNS. Oligodendrocytes arise from oligodendrocyte precursor cells (OPCs) and are found only in the CNS. [0003] Myelin is the main component of myelin, which is a layer of membrane (composed of Schwann cells and myelin cell membranes) wrapped around the ax...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/10C12N5/079C12N5/0786C12P21/02C07K14/475A61K35/44A61P25/00A61P25/28A61P9/10A61P25/16A61P25/02
CPCC12N5/069C12N5/0692C12N5/0622C12N5/0645C12P21/02C07K14/4756A61K35/44A61P25/00A61P25/28A61P9/10A61P25/16A61P25/02C12N2510/00C12N2502/28C12N2502/99C12N2501/13
Inventor 顾雨春张会远
Owner 呈诺再生医学科技(北京)有限公司
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