Method for extracting and separating extracellular secretions

Abstract

The invention discloses a method for extracting and separating extracellular secretions. The method comprises the following operation steps of placing target cells into a selected culture medium, obtaining the target cells 72 hours after cell culture, and conducting centrifugation to obtain a cell culture medium supernatant; centrifuging the obtained cell culture medium supernatant through a low-temperature centrifuge at 4 DEG C and 6,000 xg for 30 minutes, transferring the supernatant in a centrifuge tube to a new tube for later use after the centrifugation is finished, and discarding the precipitate; sucking the supernatant through a disposable syringe, and slowly dripping the supernatant to form a cell filter membrane with the specification of 0.22 microns for filtration; placing the cell culture medium into an ultracentrifuge to carry out ultra-speed centrifugation at 4 DEG C and 100,000 xg for 70 minutes, removing a supernatant, and keeping a precipitated part after the centrifugation is finished; washing and re-suspending the precipitate through a PBS buffer solution, putting the mixed liquid into the ultracentrifuge again to carry out ultra-speed centrifugation at 4 DEG C and 100,000 xg for 70 minutes, carefully sucking out a supernatant and keeping a precipitated part after the centrifugation is finished; using the PBS buffer solution for resuspending and precipitationto obtain a precipitate, namely an extracted and separated exosome product.

Description

technical field [0001] The invention relates to the technical field of exosome extraction and separation, in particular to a method for extraction and separation of cellular exosomes. Background technique [0002] At present, the extraction methods of exosomes mainly include ultracentrifugation, density gradient centrifugation, polymer precipitation and immunomagnetic bead method. Existing exosome extraction methods are difficult to balance in terms of operability, cost, extraction efficiency and product purity. Usually, the operation is simple, and the purity of exosomes extracted by less time-consuming methods is difficult to guarantee. The extraction methods that can obtain high-purity exosomes are often time-consuming, complicated to operate, and require expensive equipment support. Contents of the invention [0003] The purpose of the present invention is to provide a method for extracting and separating cellular exosomes, so as to solve the problems raised in the ab...

AI Technical Summary

Problems solved by technology

Existing exosome extraction methods are difficult to balance in terms of operability, cost, extraction efficiency and product purity. Usually, the operation is simple, and the purity of exosomes extracted by less time-consuming methods is difficult to guarantee.

However, extraction methods that can obtain high-purity exosomes are often time-consuming, complicated to operate, and require expensive equipment support