Addition of thrombin reduces the recovery of extracellular vesicles from blood plasma
Authors
Anush Arakelyan
Section on Intercellular Interactions, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA
Wendy Fitzgerald
Section on Intercellular Interactions, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA
Murad Vagida
Laboratory of Atherothrombosis, Moscow State University of Medicine and Dentistry, Moscow, Russia
Elena Vasilieva
Laboratory of Atherothrombosis, Moscow State University of Medicine and Dentistry, Moscow, Russia
Leonid Margolis
Section on Intercellular Interactions, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA
Jean-Charles Grivel
Sidra Medical and Research Center, Doha, Qatar
Extracellular vesicles (EVs) are widely studied as a system of intercellular communication, as markers of various diseases, as well as a vehicle for delivery of various bioactive molecules to various cells. Investigation of EVs’ structure and function requires their isolation and precise quantification. However, in the current literature, there are significant discrepancies in the estimated numbers of EVs in different body fluids. In part, this discrepancy is due to the difference in EVs isolation protocols used by different investigators. A common protocol that includes ExoQuick™ is often used to isolate EVs from body fluids and culture medium. Here, we show that in the case of isolation of EVs from blood, thrombin should be omitted from the protocol as clots formed due to the thrombin-triggered coagulation may entrap many EVs thus leading to the underestimation of their numbers.
Arakelyan, A., Fitzgerald, W., Vagida, M., Vasilieva, E., Margolis, L., & Grivel, J.-C. (2016). Addition of thrombin reduces the recovery of extracellular vesicles from blood plasma. Journal of Circulating Biomarkers, 5(1). https://doi.org/10.33393/jcb.2016.2063