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. 2024 Jul 4;22:623. doi: 10.1186/s12967-024-05249-w

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Characterization of extracellular vesicles (EVs) from Simpson Golabi Behmel Syndrome (SGBS) cells. The differentiation of pre-SGBS cells (a) into mature, lipid-laden SGBS cells was induced (b), after which EVs were isolated from culture medium by differential steps of ultracentrifugation. Scanning electron microscopy of EVs from mature SGBS cells (c) reveals good sample purity, and transmission electron micrograph a high-magnification image of a typical EV (d). The presence of EV-markers in EV samples was confirmed by fluorescent labelling of CD63 and CD9 in confocal microscopy (e). The presence of adipocyte-derived material (fatty acid binding protein 4 (FABP4) and adiponectin), tumor susceptibility 101 (TSG101), CD63, CD9, programmed cell death 6 interacting protein (Alix) and β-actin, as well as the absence of calnexin were further analyzed by Western Blotting from mature SGBS cell EV samples (f). Nanoparticle tracking analysis (NTA) of EV samples from pre- and mature SGBS cells revealed concentration (g) and size distribution (h) of secreted particles. NTA results are presented as mean + SEM, from 4 independent experiments. *p = 0.021 (Mann–Whitney U test). Differences in FA profiles of pre- and mature cells and their EVs were determined from total lipids with gas chromatography–mass spectrometry (i). Results are presented as percentage differences, calculated by subtracting the mol-% of each FA in the pre-group from the mol-% in the mature group. Red indicates an increase in the mature group, while blue indicates a decrease. DMA plasmalogen alkenyl chain-derived dimethyl acetal derivative, SFA saturated fatty acid, MUFA monounsaturated fatty acid, PUFA polyunsaturated fatty acid, unsaturated FA (UFA) = MUFA + PUFA. *p ≤ 0.05 Mann–Whitney U test vs. control. Percentages of selected FAs in pre- and mature cells and secreted EVs, presented as mean mol-% (j). The supervised discriminant analysis of FA proportions in pre- and mature SGBS cells, as well as their EVs (k)