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Human adipose mesenchymal stem cell exosome with high expression of IL-10 for treating myocardial infarction and use of exosome

A technique for stem cells and myocardial infarction, which is applied to genetically modified cells, cells modified by introducing foreign genetic material, animal cells, etc. Damage-relieving effect

Pending Publication Date: 2020-11-17
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the retention rate of exogenous IL-10 in the damaged area and the body's own rejection will affect its function. How to make IL-10 play a stable role in the damaged area of ​​​​myocardial infarction remains to be further studied

Method used

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  • Human adipose mesenchymal stem cell exosome with high expression of IL-10 for treating myocardial infarction and use of exosome
  • Human adipose mesenchymal stem cell exosome with high expression of IL-10 for treating myocardial infarction and use of exosome
  • Human adipose mesenchymal stem cell exosome with high expression of IL-10 for treating myocardial infarction and use of exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The purpose of this example is to isolate and culture human adipose-derived mesenchymal stem cells.

[0038] 1. Materials

[0039] Prepare human fat (normal human fat discarded after surgery, signed informed consent), PBS, BSA, type IV collagenase, fetal bovine serum, CD29 + , CD44 + , CD90 + , CD34 - and CD45 - of monoclonal antibodies.

[0040] 2. Method

[0041] 2.1 Isolation of human adipose-derived mesenchymal stem cells

[0042] (1) Collect about 15 mL of adipose tissue under sterile conditions, and remove the connective tissue capsule and blood vessels on the surface of the adipose tissue;

[0043] (2) Quickly cut into paste, add 3 mL of exosome-free medium, which contains 0.06-0.16% type IV collagenase and 0.015-0.02% BSA at final concentration , 37℃, constant temperature, 120r / min, oscillating digestion for 30-35min, observe the digestion status of the tissue at any time, take it out and mix it every 5-8min during the period, after the digestion is term...

Embodiment 2

[0061] The purpose of this example is to isolate and purify human adipose-derived mesenchymal stem cell exosomes, including the following steps:

[0062] 1. Material method

[0063] Prepare human fat (normal body fat discarded after surgery, signed informed consent), PBS, BSA, type IV collagenase, fetal bovine serum, CD9, CD63, CD81, α-actinin-4 and CD40 monoclonal antibodies and Dimethyloxalylglycine.

[0064] 2. Collection of human adipose-derived mesenchymal stem cell exosomes

[0065] (1) Collect human adipose-derived mesenchymal stem cells, inoculate them, and use exosome-free serum medium at 37°C, 5% CO 2 Start culturing under the conditions;

[0066] (2) After culturing for 12 hours, replace it with a serum medium containing 300 μmol / L dimethyloxalylglycine exosomes from human adipose-derived mesenchymal stem cells, continue culturing for 36 hours, and collect the cell culture medium;

[0067] (3) The cell culture solution obtained in step (2) was subjected to diffe...

Embodiment 3

[0082] The purpose of this example is to culture h-MSCs cell line.

[0083] 1. Materials

[0084] Prepare human fat (normal human fat discarded after surgery, signed informed consent), PBS, BSA, type IV collagenase, fetal bovine serum, CD29 + , CD44 + , CD90 + , CD34 - and CD45 - of monoclonal antibodies.

[0085] 2. Passage of h-MSCs

[0086] (1) Collect adipose tissue according to the method shown in Example 1, digest it with the digestion conditions of 0.1% type IV collagenase + 0.02% BSA group, and culture the cells according to the method described in Example 1 until 80% to 90% confluence , digested with 0.25% trypsin-0.02% EDTA, and centrifuged to obtain cell pellets;

[0087] (2) Resuspend the cell pellet in fresh exosome-free medium, passage according to 1:2~4, and culture to P10, each generation of cells is named P1-h-MSCs, P2-h-MSCs, P3-h-MSCs, respectively. h-MSCs, P4-h-MSCs, P5-h-MSCs, P6-h-MSCs, P7-h-MSCs, P8-h-MSCs, P9-h-MSCs and P10-h-MSCs;

[0088] (3)...

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Abstract

The invention provides a human adipose mesenchymal stem cell exosome with high expression of IL-10 for treating myocardial infarction and use of the exosome, and relates to the technical field of biology. The invention provides the human adipose mesenchymal stem cell exosome with high IL-10 expression for treating myocardial infarction. The human adipose mesenchymal stem cell exosome is secreted by human adipose mesenchymal stem cells with over-expression of IL-10. According to the present invention, the human adipose mesenchymal stem cell exosome has the advantages that the human adipose mesenchymal stem cell exosome with over-expression of IL-10 is prepared by culturing the human adipose mesenchymal stem cells with over-expression of IL-10, and the human adipose mesenchymal stem cell exosome with over-expression of IL-10 contains a large amount of IL-10 and other various RNAs and cell factors, so that infarcted myocardial cells are effectively repaired, the heart function is enhanced, the compensatory myocardial load is reduced, and the heart failure is relieved; and a treatment medicine or a treatment strategy is provided for myocardial infarction.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to an exosome from human adipose-derived mesenchymal stem cells overexpressing IL-10 and its use. Background technique [0002] Coronary atherosclerotic heart disease is due to abnormal lipid metabolism. The lipids in the blood deposit on the smooth arterial intima, and gradually accumulate to form white plaques, causing vascular stenosis or blockage, which can lead to severe Coronary artery occlusion, blood flow interruption, partial necrosis of myocardium due to severe persistent ischemia, resulting in myocardial infarction. After myocardial infarction, some myocardial cells are difficult to repair due to ischemia and hypoxia, resulting in reduced cardiac function and increased compensatory myocardial load, which will lead to serious complications such as heart failure over time. During the onset of coronary heart disease, the coronary artery will be blocked for 20-30 minutes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/10C12N15/85C12N15/24A61K35/28A61P9/10
CPCC12N5/0667C12N15/85C07K14/5428A61K35/28A61P9/10C12N2509/00C12N2510/00C12N2800/107
Inventor 陈良万柴天赐
Owner FUJIAN MEDICAL UNIV UNION HOSPITAL
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