An imaging flow cytometry-based methodology for the analysis of single extracellular vesicles in unprocessed human plasma.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
29 06 2022
29 06 2022
Historique:
received:
18
02
2022
accepted:
18
05
2022
entrez:
29
6
2022
pubmed:
30
6
2022
medline:
2
7
2022
Statut:
epublish
Résumé
Extracellular vesicles (EVs) are tissue-specific particles released by cells containing valuable diagnostic information in the form of various biomolecules. To rule out selection bias or introduction of artefacts caused by EV isolation techniques, we present a clinically feasible, imaging flow cytometry (IFCM)-based methodology to phenotype and determine the concentration of EVs with a diameter ≤400 nm in human platelet-poor plasma (PPP) without prior isolation of EVs. Instrument calibration (both size and fluorescence) were performed with commercial polystyrene beads. Detergent treatment of EVs was performed to discriminate true vesicular events from artefacts. Using a combination of markers (CFSE & Tetraspanins, or CD9 & CD31) we found that >90% of double-positive fluorescent events represented single EVs. Through this work, we provide a framework that will allow the application of IFCM for EV analysis in peripheral blood plasma in a plethora of experimental and potentially diagnostic settings. Additionally, this direct approach for EV analysis will enable researchers to explore corners of EVs as cellular messengers in healthy and pathological conditions.
Identifiants
pubmed: 35768629
doi: 10.1038/s42003-022-03569-5
pii: 10.1038/s42003-022-03569-5
pmc: PMC9243126
doi:
Substances chimiques
Biomarkers
0
Polystyrenes
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
633Informations de copyright
© 2022. The Author(s).
Références
Vaccines (Basel). 2019 Aug 28;7(3):
pubmed: 31466253
J Invest Dermatol. 2017 Aug;137(8):1622-1629
pubmed: 28648952
EBioMedicine. 2019 Jul;45:351-361
pubmed: 31229437
J Thromb Haemost. 2014 May;12(5):614-27
pubmed: 24618123
J Extracell Vesicles. 2019 Nov 29;9(1):1697028
pubmed: 31839906
Curr Protoc Cytom. 2018 Oct;86(1):e43
pubmed: 30168659
J Extracell Vesicles. 2015 Dec 31;4:30087
pubmed: 26725829
J Extracell Vesicles. 2019 Mar 21;8(1):1588555
pubmed: 30949309
J Clin Invest. 2016 Apr 1;126(4):1139-43
pubmed: 27035805
Int J Biol Sci. 2018 Apr 30;14(6):633-643
pubmed: 29904278
Chem Rev. 2018 Feb 28;118(4):1917-1950
pubmed: 29384376
J Extracell Vesicles. 2013 May 27;2:
pubmed: 24009894
J Clin Invest. 2017 Apr 3;127(4):1375-1391
pubmed: 28319051
Nat Rev Nephrol. 2017 Sep;13(9):545-562
pubmed: 28736435
J Thromb Haemost. 2014 Jul;12(7):1182-92
pubmed: 24818656
J Extracell Vesicles. 2020 Sep 24;9(1):1816641
pubmed: 33062218
Am J Physiol Renal Physiol. 2014 Jun 1;306(11):F1251-9
pubmed: 24694589
Int J Mol Sci. 2020 Oct 14;21(20):
pubmed: 33066349
Sci Rep. 2014 Jun 10;4:5237
pubmed: 24913598
J Cell Biol. 2013 Feb 18;200(4):373-83
pubmed: 23420871
Front Immunol. 2018 Jul 06;9:1583
pubmed: 30034401
J Thromb Haemost. 2018 Jul;16(7):1423-1436
pubmed: 29781099
J Am Soc Nephrol. 2021 May 3;32(5):1210-1226
pubmed: 33782168
Mol Cancer. 2019 Mar 30;18(1):59
pubmed: 30925927
Cytometry A. 2013 Feb;83(2):242-50
pubmed: 23125136
J Extracell Vesicles. 2020 Nov;10(1):e12028
pubmed: 33613872
Cell Mol Life Sci. 2018 Aug;75(15):2873-2886
pubmed: 29441425
Methods. 2017 Jan 1;112:55-67
pubmed: 27721015
J Extracell Vesicles. 2020 Feb 3;9(1):1713526
pubmed: 32128070
Cytometry A. 2014 Sep;85(9):756-70
pubmed: 24903900
Blood. 2011 Jan 27;117(4):e39-48
pubmed: 21041717
J Extracell Vesicles. 2019 Mar 21;8(1):1587567
pubmed: 30949308
Circ Res. 2017 Sep 29;121(8):920-922
pubmed: 28963190
Biomed Res Int. 2018 Jan 30;2018:8545347
pubmed: 29662902
J Extracell Vesicles. 2014 Feb 04;3:
pubmed: 24511371
J Extracell Vesicles. 2016 Oct 31;5:32945
pubmed: 27802845