Employing Flow Cytometry to Extracellular Vesicles Sample Microvolume Analysis and Quality Control

Frontiers in Cell and Developmental Biology
Joana MaiaBruno Costa-Silva

Abstract

Extracellular Vesicles (EVs), membrane vesicles released by all cells, are emerging mediators of cell-cell communication. By carrying biomolecules from tissues to biofluids, EVs have attracted attention as non-invasive sources of clinical biomarkers in liquid biopsies. EVs-based liquid biopsies usually require EVs isolation before content analysis, which frequently increases sample volume requirements. We here present a Flow Cytometry (FC) strategy that does not require isolation or concentration of EVs prior to staining. By doing so, it enables population analysis of EVs in samples characterized by challenging small volumes, while reducing overall sample processing time. To illustrate its application, we performed longitudinal non-lethal population analysis of EVs in mouse plasma and in single-animal collections of murine vitreous humor. By quantifying the proportion of vesicular particles in purified and non-purified biological samples, this method also serves as a precious tool to quality control isolates of EVs purified by different protocols. Our FC strategy has an unexplored clinical potential to analyze EVs in biofluids with intrinsically limited volumes and to multiply the number of different analytes in EVs that can be...Continue Reading

References

Dec 30, 2003·Cytometry. Part B, Clinical Cytometry·Abe SchwartzE Fernandez-Repollet
Aug 25, 2004·Proceedings of the National Academy of Sciences of the United States of America·Trairak PisitkunMark A Knepper
May 24, 2005·International Immunology·Marie-Pierre CabyChristian Bonnerot
Aug 21, 2007·Biotechnology and Bioengineering·Christopher J Roberts
Dec 5, 1995·Biotechnology and Bioengineering·M C ManningE Sheftert
May 14, 2010·Investigative Ophthalmology & Visual Science·Uttio Roy ChowdhuryMichael P Fautsch
Jan 8, 2011·Biological & Pharmaceutical Bulletin·Yuko OgawaRyohei Yanoshita
Apr 13, 2011·Journal of Thrombosis and Haemostasis : JTH·W L ChandlerJ F Tait
Oct 26, 2011·Nanomedicine : Nanotechnology, Biology, and Medicine·Esther N M Nolte-'t HoenMarca H M Wauben
Mar 8, 2012·Journal of Thrombosis and Haemostasis : JTH·E van der PolT G van Leeuwen
Sep 7, 2013·Journal of Extracellular Vesicles·Jason Webber, Aled Clayton
Aug 7, 2014·Clinical Proteomics·Krishna R MurthyAkhilesh Pandey
Feb 13, 2015·Analytical Chemistry·Daniele MaioloDoris Ricotta
Feb 18, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Tom Groot KormelinkMarca H M Wauben
May 20, 2015·Nature Cell Biology·Bruno Costa-SilvaDavid Lyden
May 29, 2015·PloS One·Jessica M SkeieVinit B Mahajan
Nov 3, 2015·Nature·Ayuko HoshinoDavid Lyden
Feb 10, 2016·Proceedings of the National Academy of Sciences of the United States of America·Joanna KowalClotilde Théry
Mar 25, 2016·Scientific Reports·Lucia PaoliniPaolo Bergese
Apr 2, 2016·The Journal of Clinical Investigation·Raghu Kalluri
Jan 6, 2017·Current Protocols in Cytometry·Lili Wang, Robert A Hoffman
Aug 24, 2017·Lab on a Chip·Jose C Contreras-NaranjoVictor M Ugaz
Sep 25, 2017·Advances in Experimental Medicine and Biology·Claudia Arenaccio, Maurizio Federico
Feb 25, 2018·Pharmacology & Therapeutics·Maarten P BebelmanS Rubina Baglio
Mar 9, 2018·Frontiers in Cell and Developmental Biology·Joana MaiaBruno Costa-Silva
May 9, 2018·PloS One·Emily N PrendergastKate Lawrenson
May 19, 2018·Translational Vision Science & Technology·Yuanjun ZhaoJeffrey M Sundstrom
May 22, 2018·Journal of Thrombosis and Haemostasis : JTH·S F W M LibregtsM H M Wauben
Aug 29, 2018·Journal of Extracellular Vesicles·Shang-Chun GuoHelen Dawn
Sep 6, 2018·The New England Journal of Medicine·Ravi ShahJane E Freedman
Sep 21, 2018·American Society of Clinical Oncology Educational Book·Xiaoliang WuPatrick C Ma

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Methods Mentioned

BETA
biopsies
chips
ELISA
Flow Cytometry
Size Exclusion Chromatography
blood collection
Fluorescence

Software Mentioned

Nanosight
ExoQuick
NTA
FlowJo
GraphPad
GraphPad Prism

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