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J Extracell Vesicles. 2017 Apr 10;6(1):1305677. doi: 10.1080/20013078.2017.1305677. eCollection 2017.

Characterisation of adipocyte-derived extracellular vesicle subtypes identifies distinct protein and lipid signatures for large and small extracellular vesicles.

Author information

1
INSERM U1063, Oxidative stress and metabolic pathologies, Angers University, Pointe à Pitre, France.
2
Adaptation to Tropical Climate and Exercise Laboratory, EA3596, University of the French West Indies, Pointe-à-Pitre, Guadeloupe, France.
3
GABI, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, France.
4
EXCILONE, Elancourt, France.
5
MICALIS Institute, INRA, AgroParisTech, PAPPSO, Université Paris-Saclay, Jouy-en-Josas, France.
6
Center for Medical Research, Medical University of Graz, Graz, Austria.
7
Omics Center Graz, Graz, Austria.
8
SCIAM, Angers University, Angers, France.

Abstract

Extracellular vesicles (EVs) are biological vectors that can modulate the metabolism of target cells by conveying signalling proteins and genomic material. The level of EVs in plasma is significantly increased in cardiometabolic diseases associated with obesity, suggesting their possible participation in the development of metabolic dysfunction. With regard to the poor definition of adipocyte-derived EVs, the purpose of this study was to characterise both qualitatively and quantitatively EVs subpopulations secreted by fat cells. Adipocyte-derived EVs were isolated by differential centrifugation of conditioned media collected from 3T3-L1 adipocytes cultured for 24 h in serum-free conditions. Based on morphological and biochemical properties, as well as quantification of secreted EVs, we distinguished two subpopulations of adipocyte-derived EVs, namely small extracellular vesicles (sEVs) and large extracellular vesicles (lEVs). Proteomic analyses revealed that lEVs and sEVs exhibit specific protein signatures, allowing us not only to define novel markers of each population, but also to predict their biological functions. Despite similar phospholipid patterns, the comparative lipidomic analysis performed on these EV subclasses revealed a specific cholesterol enrichment of the sEV population, whereas lEVs were characterised by high amounts of externalised phosphatidylserine. Enhanced secretion of lEVs and sEVs is achievable following exposure to different biological stimuli related to the chronic low-grade inflammation state associated with obesity. Finally, we demonstrate the ability of primary murine adipocytes to secrete sEVs and lEVs, which display physical and biological characteristics similar to those described for 3T3-L1. Our study provides additional information and elements to define EV subtypes based on the characterisation of adipocyte-derived EV populations. It also underscores the need to distinguish EV subpopulations, through a combination of multiple approaches and markers, since their specific composition may cause distinct metabolic responses in recipient cells and tissues.

KEYWORDS:

Extracellular vesicles; adipocytes; exosomes; large EVs; microvesicles; small EVs

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