Isolation and Characterization of Small Extracellular Vesicles from Porcine Blood Plasma, Cerebrospinal Fluid, and Seminal Plasma

0506877 - ÚŽFG 2020 RIV US eng J - Článek v odborném periodiku
Kupcová Skalníková, Helena - Bohuslavová, Božena - Turnovcová, Karolína - Juhásová, Jana - Juhás, Štefan - Rodinová, Marie - Vodička, Petr
Isolation and Characterization of Small Extracellular Vesicles from Porcine Blood Plasma, Cerebrospinal Fluid, and Seminal Plasma.
Proteomes. Roč. 7, č. 2 (2019), č. článku 17. E-ISSN 2227-7382
Grant CEP: GA MŠMT(CZ) LO1609
Institucionální podpora: RVO:67985904
Klíčová slova: extracellular vesicle * exosome * body fluid
Obor OECD: Cell biology
Způsob publikování: Omezený přístup
https://asep.lib.cas.cz/arl-cav/cs/csg/?repo=crepo1&key=10642646871

Extracellular vesicles (EVs) are a highly attractive subject of biomedical research as possible carriers of nucleic acid and protein biomarkers. EVs released to body fluids enable indirect access to inner organs by so-called liquid biopsies. Obtaining a high-quality EV sample with minimum contaminants is crucial for proteomic analyses using LC-MS/MS or other techniques. However, the EV content in various body fluids largely differs, which may hamper subsequent analyses. Here, we present a comparison of extracellular vesicle yields from blood plasma, cerebrospinal fluid, and seminal plasma using an experimental pig model. Pigs are widely used in biomedical research as large animal models with anatomy and physiology close to those of humans and enable studies (e.g., of the nervous system) that are unfeasible in humans. EVs were isolated from body fluids by differential centrifugation followed by ultracentrifugation. EVs were characterized according to protein yields and to the quality of the isolated vesicles (e.g., size distribution, morphology, positivity for exosome markers). In our experimental setting, substantial differences in EV amounts were identified among body fluids, with the seminal plasma being the richest EV source. The yields of pellet proteins from ultracentrifugation of 1 mL of porcine body fluids may help to estimate body fluid input volumes to obtain sufficient samples for subsequent proteomic analyses.
Trvalý link: http://hdl.handle.net/11104/0298009