Characterisation of adipocyte-derived extracellular vesicles released pre- and post-adipogenesis
Connolly, Katherine D.
Guschina, Irina A.
Draman, Mohd Shazli
Von Ruhland, Christopher
Rees, D. Aled
MetadataShow full item record
Extracellular vesicles (EVs) are submicron vesicles released from many cell types, including adipocytes. EVs are implicated in the pathogenesis of obesity-driven cardiovascular disease, although the characteristics of adipocyte-derived EVs are not well described. We sought to define the characteristics of adipocyte-derived EVs before and after adipogenesis, hypothesising that adipogenesis would affect EV structure, molecular composition and function. Using 3T3-L1 cells, EVs were harvested at day 0 and day 15 of differentiation. EV and cell preparations were visualised by electron microscopy and EVs quantified by nanoparticle tracking analysis (NTA). EVs were then assessed for annexin V positivity using flow cytometry; lipid and phospholipid composition using 2D thin layer chromatography and gas chromatography; and vesicular protein content by an immuno-phenotyping assay. Pre-adipogenic cells are connected via a network of protrusions and EVs at both time points display classic EV morphology. EV concentration is elevated prior to adipogenesis, particularly in exosomes and small microvesicles. Parent cells contain higher proportions of phosphatidylserine (PS) and show higher annexin V binding. Both cells and EVs contain an increased proportion of arachidonic acid at day 0. PREF-1 was increased at day 0 whilst adiponectin was higher at day 15 indicating EV protein content reflects the stage of adipogenesis of the cell. Our data suggest that EV production is higher in cells before adipogenesis, particularly in vesicles <300 nm. Cells at this time point possess a greater proportion of PS (required for EV generation) whilst corresponding EVs are enriched in signalling fatty acids, such as arachidonic acid, and markers of adipogenesis, such as PREF-1 and PPARγ.
Journal of Extracellular Vesicles
Connolly, K.D., Guschina, I.A., Yeung, V., Clayton, A., Draman, M.S., Von Ruhland, C., Ludgate, M., James, P.E. and Rees, D.A.(2015) 'Characterisation of adipocyte-derived extracellular vesicles released pre-and post-adipogenesis', Journal of Extracellular Vesicles, 4.
This article was published in Journal of Extracellular Vesicles on 24th November 2014 (online),available at http://dx.doi.org/10.3402/jev.v4.29159
RightsCreative Commons Attribution Non-Commercial 3.0 Unported License
Except where otherwise noted, this item's license is described as Creative Commons Attribution Non-Commercial 3.0 Unported License
Showing items related by title, author, subject and abstract.
Connolly, Katherine; Wadey, Rebecca; Matthew, Donna; Johnson, Errin; Rees, D. Aled; James, Philip (Oxford University Press, 2018-07-16)Adipocyte-derived extracellular vesicles (EVs) may serve as novel endocrine mediators of adipose tissue and impact upon vascular health. However, it is unclear whether adipocyte-derived EVs are present in the human ...
Bariatric surgery is accompanied by changes in extracellular vesicle-associated and plasma Fatty Acid Binding Protein 4 Witczak, Justyna; Min, Thinzar; Prior, Sarah L.; Stephens, Jeffrey W.; James, Philip; Rees, Aled (Springer link, 2017-08-19)BACKGROUND: Bariatric surgery markedly reduces fat mass with beneficial effects on cardiometabolic health but the mechanisms involved are not fully understood. Extracellular vesicles (EVs) are secreted by a variety of ...
Ahmeed, Al Maamari Rahima Ali (Cardiff Metropolitan University, 2010)Conjugated linoleic acid (CLA) is a dietary polyunsaturated fatty acid. It is a mixture of positional and geometrical (cis/tran) isomers of octadecadienoic fatty acids derived from linoleic acid. CLA is naturally occurring ...