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. 2020 Mar 14;9(3):721. doi: 10.3390/cells9030721

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Characterization of serum and urine EVs from patients with CCA, PSC, UC, and healthy controls. In order to validate the protocol for EVs isolation, we used blood serum and urine from healthy individuals. (A) TEM images of blood serum (left) and urine (right) EVs from healthy individuals showcasing the typical round shape (~150 nm) and morphology. (B) Representative immunoblots of the EV markers CD63 and CD81 (positive controls) and GRP78 (negative control) from EVs isolated from serum (left) and urine (right) of healthy individuals that indicate an enrichment of EV markers and a complete absence of the endoplasmic reticulum (ER) marker GRP78, compared to total serum and whole cell extracts (WCEs) of normal human cholangiocytes (NHC). (C) Nanoparticle tracking analysis (NTA) of serum (up) and urine (down) EVs revealing no differences in EV concentration between CCA, PSC, UC, and healthy individuals and a similar EV mode (~180 nm).