A Rapid Method for the Separation and Enrichment of Exosomes from Tissues / 中国生物化学与分子生物学报

ABSTRACT

The study demonstrates a quick approach for isolating exosomes with substantial concentration. To achieve quality and substantial concentration and purification of exosomes, the target tissue wasmechanically chopped and introduced to tissue digesting enzyme for tissue digestion and filtration. Thetissue cell solution was then subjected to differential centrifugation, ultra-separation, SEC exclusion, andultrafiltration. The protein contents of enzymatic treatment of dissociated tissue were higher as compared toprotein elution of exosome tissue dissociation method. The nanoparticles were traced and seen using atransmission electron microscope after enrichment of exudate to mouse heart, liver, kidney, human coloncancer, human breast cancer, and atherosclerotic tissues. The finding of the study demonstrated that thediameter of the exocrine body was within 30-150 nm, and the structure was obvious and distinct. Westernblots analysis showed that CD9, Alix, and CD63 expression were high, whereas, the TSG101 expressionwas low but calnexin was negative. However, in comparison to other ways, whole process takes only 4-5hours and saves time for quantification and functional analysis of exosomes. The isolated exosomes hashigher purification with less soluble heteroprotein contamination. Advances in isolation exosomes frommicro tissue samples can be employed for nanoparticle size tracking, Western blotting, transmission electron microscopy, and transcriptomic analysis for future studies.