Extraction method of cell exosome

Abstract

The invention discloses an extraction method of cell exosome. The method is obtained by improving the prior art, a culture medium after cell culture is centrifuged to remove impurities, a culture supernatant is separated out and collected, vacuum cold ice drying is conducted on the clarified culture solution, and 70mL of buffer solution is added into the dry powder to facilitate cell fusion, and aculture medium with relatively high purit is obtained; and centrifugation is carried out in a centrifugal machine after heating, and isopyknic precooled Thris-saturated acid, a precooled 0.1 mol / L ammonium acetate methanol solution with the volume of 5 times and an exosome extracting solution with the volume of 5 times, and a 100% ethanol solution with the volume of 5 times and an exosome extracting solution with the volume of 5 times are respectively added, and refusion of a cell centrifugation product is facilitated. During operation, centrifugation conditions are strictly controlled, expensive extraction methods such as ultrafiltration centrifugation and the like are not needed, exosome precipitate extracted through centrifugation is higher in purity, and the extraction time is saved.

Description

Technical field [0001] The present invention belongs to the extraction of the extraction of the extracellular body, and more particularly, the present invention relates to a method of extracting a cell exosomass. Background technique [0002] Subjects refers to a small membrane (30-150 nm) containing complex RNA and protein. Today, it refers to a disk-shaped vesicle in diameter at 40-100 nm. In 1983, the primary bodies were found in the sheep net to the red cells, and 1987 Johnstone was named "EXOSOME". A variety of cells can secrete an epistem in a normal and pathological state. It is mainly derived from the polycycosomal particles formed by intracellular lysosomas, which are released to the extracellular matrix after fusion of the polycyclic bubbles and the cell membrane. All cultured cell types can secrete the epistemis, and the outer body is naturally present in the body fluid, including blood, saliva, urine, cerebrospinal fluid and milk. The accurate molecular mechanism of "...

AI Technical Summary

Problems solved by technology

There are many problems in using PEG to precipitate exosomes: such as low purity and recovery, more impurities (false positives), uneven particle size, generation of difficult-to-remove polymers, mechanical force or chemical additives such as Tween-20 Will destroy exosomes, etc., so it is easy to be questioned when publishing an article

However, the method of sucrose gradient centrifugation is relatively time-consuming, and ultrafiltration centrifugation requires constant replacement of filter membranes.