Journal
JOURNAL OF EXTRACELLULAR VESICLES
Volume 5, Issue -, Pages -Publisher
TAYLOR & FRANCIS LTD
DOI: 10.3402/jev.v5.30829
Keywords
saliva; exosome; EVs; microRNA; diagnosis
Categories
Funding
- MEXT (Ministry of Education, Culture, Sports, Science and Technology)
- Vehicle Racing Commemorative Foundation
Ask authors/readers for more resources
Diagnostic methods that focus on the extracellular vesicles (EVs) present in saliva have been attracting great attention because of their non-invasiveness. EVs contain biomolecules such as proteins, messenger RNA (mRNA) and microRNA (miRNA), which originate from cells that release EVs, making them an ideal source for liquid biopsy. Although there have been many reports on density-based fractionation of EVs from blood and urine, the number of reports on EVs from saliva has been limited, most probably because of the difficulties in separating EVs from viscous saliva using density gradient centrifugation. This article establishes a protocol for the isolation of EVs from human saliva using density gradient centrifugation. The fractionated salivary EVs were characterized by atomic force microscopy, western blot and reverse transcription polymerase chain reaction. The results indicate that salivary EVs have a smaller diameter (47.8 +/- 12.3 nm) and higher density (1.11 g/ml) than EVs isolated from conditioned cell media (74.0 +/- 23.5 nm and 1.06 g/ml, respectively). Additionally, to improve the throughput of density-based fractionation of EVs, the original protocol was further modified by using a fixed angle rotor instead of a swinging rotor. It was also confirmed that several miRNAs were expressed strongly in the EV-marker-expressing fractions.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available