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Sci Rep. 2016 Apr 18;6:24316. doi: 10.1038/srep24316.

Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection.

Author information

1
Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 1085, Hungary.
2
Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, 1083, Hungary.
3
Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, 1085, Hungary.
4
Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, 1117, Hungary.

Abstract

Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and purification from body fluids remains challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma samples as well as human platelet concentrates. Using flow cytometry, we found that the majority of circulating particles within the size range of extracellular vesicles lacked common vesicular markers. We identified most of these particles as lipoproteins (predominantly low-density lipoprotein, LDL) which mimicked the characteristics of extracellular vesicles and also co-purified with them. Based on biophysical properties of LDL this finding was highly unexpected. Current state-of-the-art extracellular vesicle isolation and purification methods did not result in lipoprotein-free vesicle preparations from blood plasma or from platelet concentrates. Furthermore, transmission electron microscopy showed an association of LDL with isolated vesicles upon in vitro mixing. This is the first study to show co-purification and in vitro association of LDL with extracellular vesicles and its interference with vesicle analysis. Our data point to the importance of careful study design and data interpretation in studies using blood-derived extracellular vesicles with special focus on potentially co-purified LDL.

PMID:
27087061
PMCID:
PMC4834552
DOI:
10.1038/srep24316
[Indexed for MEDLINE]
Free PMC Article

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