A bead-assisted flow cytometry method for the semi-quantitative analysis of Extracellular Vesicles
Suárez, Henar (Instituto de Investigación Hospital Universitario de la Princesa)
Gámez-Valero, Ana (Institut Germans Trias i Pujol. Hospital Universitari Germans Trias i Pujol)
Reyes, Raquel (Centro de Biología Molecular Severo Ochoa)
López-Martín, Soraya (Instituto de Investigación Hospital Universitario de la Princesa)
Rodríguez, María Josefa (Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas)
Carrascosa, José L. (Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas)
Cabañas, Carlos (Centro de Biología Molecular Severo Ochoa)
Borràs i Serres, Francesc Enric (Institut Germans Trias i Pujol. Hospital Universitari Germans Trias i Pujol)
Yáñez-Mó, María (Instituto de Investigación Hospital Universitario de la Princesa)
Universitat Autònoma de Barcelona

Date:	2017
Abstract:	Most experimental approaches commonly employed for the characterization and quantitation of EVs are time consuming, require of specialized instrumentation and often are rather inaccurate. To circumvent the caveats imposed by EV small size, we used general and specific membrane markers in bead assisted flow cytometry, to provide a semi-quantitative measure of EV content in a given sample. EVs were isolated from in vitro cultured cells-conditioned medium and biological fluids by size exclusion chromatography and coupled to latex beads to allow their detection by standard flow cytometers. Our analyses demonstrate a linear correlation between EV concentration and Mean Fluorescence Intensity values in samples cleared of protein contaminants. Comparison with one of the most widespread method such as NTA, suggests a similar linear range and reliable accuracy to detect saturation. However, although detection of the different biomarkers is feasible when tested on ultracentrifugationenriched samples, protein contamination impairs quantitation of this type of samples by bead-based flow cytometry. Thus, we provide evidence that bead-assisted flow cytometry method is an accurate and reliable method for the semiquantitative bulk analysis of EVs, which could be easily implemented in most laboratories.
Grants:	Ministerio de Economía y Competitividad BFU2014-55478-R Instituto de Salud Carlos III PI13/00050 Agència de Gestió d'Ajuts Universitaris i de Recerca 2014SGR804 Ministerio de Economía y Competitividad BFU2014-54181-P Ministerio de Economía y Competitividad SAF2016-77096-R Ministerio de Ciencia e Innovación RYC-2012-11025
Note:	Altres ajuts: Fundación Ramón Areces; FPI-UAM i GEIVEX; La Marató de TV3 (482/U/2014)
Note:	Altres ajuts: FEDER/REDinREN/16/0009
Note:	Altres ajuts: GC/CES07/015
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Published in:	Scientific reports, Vol. 7 (september 2017) , p. 1-11, ISSN 2045-2322

DOI: 10.1038/s41598-017-11249-2
PMID: 28900146

11 p, 2.5 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut d'Investigació en Ciencies de la Salut Germans Trias i Pujol (IGTP)
Articles > Research articles
Articles > Published articles