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Mesenchymal stem cell source exosome based on Tongxinluo pretreatment and preparation method thereof

A technique for stem cells and exosomes, which is applied in the field of high-efficiency exosomes derived from mesenchymal stem cells and their preparation, can solve the problems of insignificant effects and the like

Active Publication Date: 2021-09-10
FUWAI HOSPITAL CHINESE ACAD OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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Problems solved by technology

The paracrine function of MSCs is an important way for them to function (Ranganath, S.H., et al., Harnessing the mesenchymalstem cell secretome for the treatment of cardiovascular disease. Cell StemCell, 2012.10(3):p.244-58.), however Clinical studies have shown that although mesenchymal stem cells can improve cardiac function after infarction through paracrine protection to a certain extent, the effect is not significant

Method used

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  • Mesenchymal stem cell source exosome based on Tongxinluo pretreatment and preparation method thereof
  • Mesenchymal stem cell source exosome based on Tongxinluo pretreatment and preparation method thereof
  • Mesenchymal stem cell source exosome based on Tongxinluo pretreatment and preparation method thereof

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Embodiment 1

[0033] Preparation method of exosomes derived from mesenchymal stem cells:

[0034] The preparation method of exosomes derived from mesenchymal stem cells: the primary rat (Sprague-Dawley rat, 60-80g) BM-MSCs were isolated by the differential attachment method, and the BM-MSCs were subcultured and expanded to the 3rd-4th generation for later use. BM-MSCs complete medium (IMDM, Gibco, USA) was pretreated with 400 μg / mL Tongxinluo for 24 hours, and the cell medium was replaced with IMDM partial medium (without adding fetal bovine serum) to continue culturing. After 48 hours, the conditioned medium was collected and ultracentrifugation was used to separate the exosomes secreted by BM-MSCs pretreated with Tongxinluo (MSC TXL -Exo). The specific steps of the ultracentrifugation method include: after collecting the conditioned medium, centrifuge at 300g for 10min to remove cells; centrifuge at 2000g for 20min to remove cell debris; high-speed centrifuge at 16500g for 30min to remov...

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Abstract

The invention provides a mesenchymal stem cell source exosome based on Tongxinluo pretreatment and a preparation method thereof. The invention provides the preparation method of the mesenchymal stem cell source exosome for up-regulating the expression level of miR-146a. The method comprises the following steps of pretreating mesenchymal stem cells by adopting Tongxinluo, and culturing the treated mesenchymal stem cells to collect the exosome secreted by the mesenchymal stem cells. According to the mesenchymal stem cell source exosome, the mesenchymal stem cell source exosome capable of promoting angiogenesis and / or myocardial repair can be prepared.

Description

technical field [0001] The present invention relates to a mesenchymal stem cell-derived exosome based on drug pretreatment and its preparation method, in particular to a high-efficiency mesenchymal stem cell-derived exosome based on drug Tongxinluo pretreatment and its preparation method. Background technique [0002] WHO statistics show that in 2016, cardiovascular disease was the leading cause of death in the world (about 17.6 million people, 32.2%). Although the current medical level has been greatly improved, acute myocardial infarction (acute myocardial infarction, AMI) is still the leading cause of death worldwide. One of the main causes of disability (GBD 2016 Mortality Collaborators. Global, regional, and national under-5 mortality, adult mortality, age-specific mortality, and life expectancy, 1970-2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet. 2017 Sep 16; 390(10100):1084-1150; Roth, G.A., et al., Global, Regional, and National Bur...

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P9/10
CPCC12N5/0662C12N5/0663C12N5/0667A61K35/28A61P9/10C12N2509/00C12N2500/76C12N2500/82
Inventor 杨跃进熊语嫣汤瑞杰
Owner FUWAI HOSPITAL CHINESE ACAD OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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