Extracellular vesicles collected from damaged tissues and cells infected with SARS-CoV-2

概要

Extracellular vesicles (EVs), which are nanosized vesicles released by cells as intercellular messengers, have a high potential as biomarkers. EVs have been reported to participate in viral transmission or modulation of pro- and anti-viral response. Further investigation of EVs from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected cells may provide clues for coronavirus disease (COVID-19) pathogenesis and therapies. In this study, I collected and characterized EVs from Calu-3 cells infected with SARS-CoV-2 in vitro. SARSCoV-2 RNA was detected in samples containing EVs.

In the second project, I focused on damaged tissues as a possible source for EV collection. EVs are usually collected from in vitro sources, such as cell culture media or biofluids, and not from tissues. Techniques enabling direct collection of EVs from tissues will extend the applications of EVs. I compared methods for separating EVs from solid liver, heart, and skeletal muscle. Compared with a precipitation method, an ultracentrifugation-based method for collection of EVs from solid tissues yielded a higher proportion of EVs positive for EV-related markers, with minimum levels of intracellular organelle-related markers. Some tissue-specific modifications, such as a sucrose cushion step, may improve the yield and purity of the collected EVs.