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Method for improving yield of mesenchymal stem cell exosomes

A technology of mesenchymal stem cells and exosomes, applied in the field of increasing the production of exosomes secreted by mesenchymal stem cells, can solve the problems of a large amount of manpower, material and financial resources, limited ability to secrete exosomes, slow down the progress of experimental research, etc. Increase, beneficial effect of treatment

Inactive Publication Date: 2019-01-11
ZHEJIANG UNIV
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Problems solved by technology

However, the ability of mesenchymal stem cells to secrete exosomes is limited, and only a small amount of exosomes can be obtained from large-scale cultured cells. The cultivation and extraction process requires a lot of manpower, material and financial resources, which may slow down the progress of experimental research. Therefore, Optimizing the stem cell culture system to increase the production of exosomes will help to improve its future clinical application value

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  • Method for improving yield of mesenchymal stem cell exosomes

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Embodiment 1

[0016] 1. MSC cell lines are routinely cultured in cell in vitro medium, supplemented with 100U / ml penicillin and 100U / ml streptomycin, once pooled, use 0.25% trypsin (w / v)-0.02% ethylenediaminetetraacetic acid (EDTA) (w / v) (Gino Biomedical Technology Co., Ltd.) digested and passaged. The composition of the cell culture medium in vitro is: sodium pyruvate 110mg / L, glutamine 20mM, fetal calf serum 10% (volume concentration), and the solvent is low-sugar DMEM medium (purchased from BI Company of the United States).

[0017] 2. Wait until the MSC grows to 70%-80% confluence, in O 2 Volume concentration 0.5%, CO 2 After culturing at 37°C for 24 hours in a mixed gas with a volume concentration of 5% (the balance is nitrogen), remove the in vitro culture medium of the cells, wash with PBS for 3 times, and replace with serum-free supplemented with 100U / ml penicillin and 100U / ml streptomycin Low-sugar DMEM medium, at 37°C in CO 2 In a mixed gas with a volume concentration of 5% and...

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Abstract

The invention provides a method for improving the yield of mesenchymal stem cell exosomes. The method comprises the following steps: inoculating a mesenchymal stem cell with stable passage to a cell in-vitro culture medium; culturing in an anaerobic environment at 37 DEG C for 24 hours; culturing in a normoxia environment at 37 DEG C for 48 hours; and picking up cultured supernate, and extractingthe exosome excreted by an ultracentrifugation method. According to the method, mesenchymal stem cells subjected to anaerobic and normoxia pretreatment can secrete more exosomes, which is beneficial to treatment of cardiovascular diseases.

Description

(1) Technical field [0001] The present invention relates to a method for secreting exosomes, in particular to a method for increasing the yield of exosomes secreted by mesenchymal stem cells. (2) Background technology [0002] At present, the incidence of myocardial infarction is increasing year by year, because it will lead to irreversible damage and loss of myocardial cells, and cause scar formation, resulting in severe ventricular remodeling, a series of complications, and eventually can develop into heart failure. Leading cause of death in vascular disease. Therefore, promoting the regeneration of myocardial cells and saving the endangered myocardium has become the central link of regeneration and repair after myocardial infarction. Although there are a variety of treatment methods for relieving heart damage, the treatment efficiency is still not high enough to significantly improve the adverse outcomes of patients after myocardial infarction. Therefore, it is necessary...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2500/02
Inventor 胡新央王建安钟淑涵李庆举
Owner ZHEJIANG UNIV