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Neural stem cell microvesicles and application thereof

A technology of neural stem cells and microvesicles, applied in nervous system cells, animal cells, vertebrate cells, etc., can solve the problems of different specific proteins, different functions or applications, difficulty in obtaining a large number of cells, etc., and achieve a strong protective effect , increase the content, the effect of uniform structure

Pending Publication Date: 2019-09-20
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, major issues such as teratoma formation, immune response, difficulty in obtaining large numbers of cells, and limited cell proliferation and differentiation hinder the routine use of these cells as a clinical treatment option
[0003] At present, there have been related studies on the function and application of exosomes derived from mesenchymal stem cells or serum, but there are not many studies on microvesicles, and the specific proteins expressed by microvesicles or exosomes from different sources are different. , are also different in function or application, and the function and application of neural stem cell-derived microvesicles have not yet been disclosed

Method used

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  • Neural stem cell microvesicles and application thereof
  • Neural stem cell microvesicles and application thereof
  • Neural stem cell microvesicles and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation, identification and protein detection of microvesicles secreted by neural stem cells

[0030] The main materials and sources used in this embodiment are as follows:

[0031] Neural stem cell culture reagents: DMEM / F12, B27, 20, bFGF, EGF, non-essential amino acids (Thermo Fisher, USA), Accutase (Stemcell, Canada), monoclonal mouse anti-HSP-70, monoclonal mouse anti- β-actin (Santa Cruz Biotechnology), HRP-goat anti-mouse IgG (Shanghai Sangong), BCA protein quantification kit (Beijing Kangwei Century Company), protein maker (Quanshijin Company), premixed chemiluminescence substrate, PVDF membrane, ECL chromogenic solution (Millipore Company), Coomassie brilliant blue staining solution / decolorization solution (Beiyuntian Company), carbon dioxide incubator company, inverted microscope, biological microscope, ultra-clean workbench, desktop centrifuge, ultra-high-speed low-temperature centrifugation machine.

[0032] The specific implementation steps o...

Embodiment 2

[0048] Example 2: Microvesicles secreted by neural stem cells deliver HSP-70 function

[0049] Main material used in embodiment 2 and source are as follows respectively:

[0050] Cardiomyocyte culture reagents: high-glucose DMEM, fetal bovine serum, trypsin (Thermo Fisher), polyclonal rabbit anti-Bcl-2, polyclonal rabbit anti-Bax (USA CST Company), H 2 o 2 (Sinopharm Company), Triptolide (APExBio Company), Annexin V / PI Apoptosis Kit (BD Company), flow cytometry, and other references to Example 1.

[0051] The specific implementation steps of Embodiment 2 of the present invention are described as follows:

[0052] Mouse cardiomyocyte culture: mouse cardiomyocytes were purchased from the Cell Bank of the Chinese Academy of Sciences, and used high

[0053] Cardiomyocytes were cultured in sugar-DMEM medium at 37°C, 5% CO 2 constant temperature cell incubator.

[0054] Detection of protein expression level by Western blot: extract proteins in cardiomyocytes according to conven...

Embodiment 3

[0056] Example 3: The effect of microvesicles secreted by neural stem cells on inhibiting cardiomyocyte apoptosis

[0057] h 2 o 2 Stimulated mouse cardiomyocytes for 3 hours to simulate the oxidative stress response produced by the heart, used as an apoptosis model of cardiomyocytes, and used to simulate myocardial ischemia-reperfusion injury in vitro, H 2 o 2 The concentration is 1000 μmol / L. This embodiment compares by being divided into several groups:

[0058] NC group: adding an equal volume of PBS to the cardiomyocyte culture medium was used as the control group;

[0059] h 2 o 2 Group: adding H to cardiomyocyte culture medium 2 o 2 apoptotic cells;

[0060] h 2 o 2 +200μg / mL group: add H to the cardiomyocyte culture medium 2 o 2 And 200 μg / mL of neural stem microvesicles;

[0061] h 2 o 2 + 500μg / mL group: add H to the cardiomyocyte culture medium 2 o 2 and 500 μg / mL neural stem microvesicles.

[0062] Cell morphology was observed, cell protein was ...

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Abstract

The invention relates to neural stem cell microvesicles and an application thereof, which belongs to the technical fields of biological materials and application; The microvesicles provided by the present invention are derived from neural stem cells, and the microvesicles specifically express and can deliver the protein HSP-70; through experiments, the application of the neural stem cell microvesicles in preparation of a drug for treating cardiomyocyte apoptosis-induced diseases such as myocardial reperfusion injury is proved, the above application is to increase the content of HSP-70 in cardiomyocytes by delivering HSP-70 through neural stem cell-derived microvesicles, so that cardiomyocyte apoptosis can be effectively inhibited; the use of the microvesicles of the present invention for the treatment of myocardial reperfusion injury not only has the same effect as stem cell therapy, but also avoids problems such as tumor formation, immunosuppression, and low survival rate during stem cell therapy, and growth factors, non-coding RNA and other cytoprotective factors are still available to help repair and regenerate damaged cells or tissues.

Description

technical field [0001] The invention relates to a neural stem cell microvesicle and its application, belonging to the technical field of biological materials and applications. Background technique [0002] Ischemic heart disease (also known as coronary heart disease) is one of the leading causes of death and disability worldwide. Timely reperfusion is currently the most effective treatment for ischemic heart disease, which can not only reduce infarct size but also prevent heart failure, but the reperfusion process itself can induce myocardial cell death, which is called myocardial reperfusion injury. Myocardial reperfusion injury is caused by excessive production of reactive oxygen species, energy metabolism disorder, neutrophil infiltration, calcium overload and vascular endothelial dysfunction, and there is still no effective treatment. Stem cell transplantation therapy is considered to be an effective method for the treatment of myocardial reperfusion injury, mainly thro...

Claims

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Application Information

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IPC IPC(8): C12N5/0797A61K38/17A61P9/00A61P9/10
CPCC12N5/0623A61K38/1709A61P9/00A61P9/10
Inventor 张磊胡嘉波高建一陈天琰
Owner JIANGSU UNIV
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