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Method for extracting thrombus exosome of patient with acute myocardial infarction and application of thrombus exosome

An acute myocardial infarction and exosome technology, applied in the field of medicine, can solve the problems of complex and inapplicable thrombus components, and achieve the effect of improving purity and reducing dosage

Active Publication Date: 2021-08-20
GUIZHOU PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, research at home and abroad is based on exosomes from a specific cell source or exosomes in circulating blood. There may be some differences from circulating blood, and the components of thrombus are complex and not a single cell line, so the current conventional exosome research system is not applicable

Method used

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  • Method for extracting thrombus exosome of patient with acute myocardial infarction and application of thrombus exosome
  • Method for extracting thrombus exosome of patient with acute myocardial infarction and application of thrombus exosome
  • Method for extracting thrombus exosome of patient with acute myocardial infarction and application of thrombus exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Extraction method of thrombus exosomes in patients with acute myocardial infarction

[0043] S1: Take adult coronary artery thrombus after myocardial infarction and put it in a 15ml centrifuge tube containing 5-10ml phosphate buffer solution and let it stand overnight for more than 6h;

[0044] S2: add trypsin 20 μg / ml and 50U / ml type II collagenase;

[0045] S3: After fully oscillating and mixing, place on a constant temperature shaker at 37°C for 4 hours until the thrombus is completely dissolved;

[0046] S4: Remove the blood in the non-coagulated state, then suck the supernatant and discard it, take 100 mg of the precipitated thrombus and put it in a 15 ml centrifuge tube equipped with 4 ml PBS for subsequent use;

[0047]S5: Centrifuge the thrombus tissue lysate after the aforementioned dissolution in a high-speed centrifuge at 4° C., 2000 g×10 min, remove residual dead cells and cell fragments in the lysate, and take the supernatant for use;

[0048] S...

Embodiment 2

[0050] Example 2 Extraction method of thrombus exosomes in patients with acute myocardial infarction

[0051] S1: Take adult coronary artery thrombus after myocardial infarction and put it in a 15ml centrifuge tube equipped with 5-10ml phosphate buffer solution and let it stand overnight;

[0052] S2: Add 10 μg / ml trypsin and 10 U / ml type II collagenase;

[0053] S3: After fully oscillating and mixing, place on a constant temperature shaker at 37°C for 4 hours until the thrombus is completely dissolved;

[0054] S4: Remove the blood in the non-coagulated state, then suck the supernatant and discard it, take 90 mg of the precipitated thrombus and put it in a 15ml centrifuge tube equipped with 4mlPBS for subsequent use;

[0055] S5: Centrifuge the thrombus tissue lysate after the aforementioned dissolution in a high-speed centrifuge at 2° C., 2000 g × 7 min, remove residual dead cells and cell fragments in the lysate, and take the supernatant for later use;

[0056] S6; Add th...

Embodiment 3

[0058] Example 3 Extraction method of thrombus exosomes in patients with acute myocardial infarction

[0059] S1: Take adult coronary artery thrombus after myocardial infarction and put it in a 15ml centrifuge tube equipped with 5-10ml phosphate buffer solution and let it stand overnight;

[0060] S2: add 200 μg / ml trypsin and 200U / ml type II collagenase;

[0061] S3: After fully oscillating and mixing, place on a constant temperature shaker at 37°C for 4 hours until the thrombus is completely dissolved;

[0062] S4: Remove the blood in the non-coagulated state, then suck the supernatant and discard it, take 110 mg of the precipitated thrombus and put it in a 15 ml centrifuge tube equipped with 4 ml PBS for subsequent use;

[0063] S5: Centrifuge the thrombus tissue lysate after the aforementioned dissolution in a high-speed centrifuge at 5° C., 2000 g×13 min, remove residual dead cells and cell fragments in the lysate, and take the supernatant for later use;

[0064] S6; Ad...

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Abstract

The invention relates to a method for extracting thrombus exosomes of patients with acute myocardial infarction and application, after thrombus is dissolved by using a method of mixing collagenase with pancreatin, exosomes in thrombus are successfully extracted by using an ultracentrifugation method, which is the first case at home and abroad, and compared with an existing method for extracting tissue exosomes by using collagenase, the method has the advantages that after the pancreatin is added, the use amount of collagenase is reduced, and the purity of the exosome is also improved.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a method and application for extracting thrombus exosomes from patients with acute myocardial infarction. Background technique [0002] Myocardial infarction is also called myocardial infarction. Myocardial infarction (myocardial infarction) is coronary artery occlusion, blood flow interruption, and partial necrosis of some myocardium due to severe persistent ischemia. [0003] Clinically, there are severe and long-lasting retrosternal pain, fever, leukocytosis, accelerated erythrocyte sedimentation rate, increased serum myocardial enzyme activity and progressive ECG changes, and arrhythmia, shock or heart failure may occur. [0004] The sudden onset of acute myocardial infarction should be detected and treated early, and pre-hospital treatment should be strengthened. The principle of treatment is to save the dying myocardium, reduce the infarct size, protect the heart function...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078A61K35/14A61P9/10
CPCC12N5/0634A61K35/14A61P9/10C12N2509/00C12N2509/10
Inventor 何尤夫吴强钱宇艾丽琼周瑜黄晶龙向淑蒙宣彤
Owner GUIZHOU PROVINCIAL PEOPLES HOSPITAL
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