Method for separating exosome from fluid shear stress perfusate

Abstract

The invention discloses a method for separating exosome from fluid shear stress perfusate. A method of performing freeze-drying, redissolution and concentration on exosome in the fluid shear stress perfusate is adopted, so that the exosome in the fluid shear stress perfusate can be concentrated by about 10 times, in the concentration course, the exosome is not damaged, depletion of the exosome isnot caused, the obtained exosome is complete in structure, high in purity and low in hybridprotein pollution, the exosome in massive cell perfusate can be extracted in one time, costly equipment is not needed, used consumptive materials are low in price, and the extraction method is simple and easy to operate.

Description

technical field [0001] The invention relates to the research field of exosomes, in particular to a method for effectively separating exosomes from fluid shear stress perfusate. Background technique [0002] Previously, in vitro research on exosomes secreted by vascular endothelial cells was limited to a normal amount of cell culture fluid, the most common being cells with a bottom area of ​​10 cm2, which only required 2 ml of culture medium. At this time, the amount of cell culture medium is small, and the concentration of exosomes in it is relatively high, so it is easy to extract. However, this type of research is limited to static cells, ignoring the fluid shear stress environment of cells in vivo. Vascular endothelial cells in the body, the innermost layer of blood vessels, are normally subject to fluid shear stress generated by blood flow. [0003] Therefore, in order to better simulate the real microenvironment in vivo, it is necessary to observe the exosome secretio...

AI Technical Summary

Problems solved by technology

However, at present, the influence of fluid shear stress on endothelial cells only involves the morphology and function of cells, and does not involve the influence of exosomes secreted by cells, and after different forms of fluid shear stress (laminar flow or oscillatory flow) intervene in cells, Differential expression of contents in secreted exosomes

The main reason is that when fluid shear stress interferes with cells, a large amount of culture medium (>100 ml) is required, in which the concentration of exosomes is relatively low and the amount of liquid is large. Traditional ultracentrifugation, ultrafiltration and affinity chromatography The method can not be effectively carried out, resulting in the research on the function of such exosomes cannot be carried out smoothly

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