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Extended periods of hyperglycemia (HG) can lead to metabolic disorders of sphingolipids (SPs) and their subsequent accumulation in cells. This accumulation can trigger a range of complications, including kidney and neurodegenerative diseases. In our study, we compared the levels of selected ceramides (CER), hexosylceramides (HexCER), and glycosphingolipids (GSLs) in potential HG biomarkers - extracellular vesicles (EVs). These EVs were derived in vitro from human β-cells cultured under both normoglycemic and high-glucose conditions (HG). We utilized Time of Flight – Secondary Ion Mass Spectrometry (ToF-SIMS) for SPs analysis. Our results confirmed that the lipid profiles of these three groups differ between large and small EVs, with some SP lipids being more enriched in EVs compared to cells. Interestingly, our study revealed that HG only regulates the lipid content from the glycosphingolipid group in relation to normoglycemia. Collectively, our findings underscore the potential applications of ToF-SIMS in characterizing the impact of different culture conditions on lipid levels. As far as we know, our study is the first which employs ToF-SIMS in analyzing the effects of HG on SP levels in EVs and their parental β-cells.

Highlights

• ToF-SIMS effectively compares glycosphingolipid content in cells and EVs.

• Analysis reveals lipid profile differences between EVs and their parent cells.

• Hyperglycemia alters glycosphingolipid content in cells and EVs per ToF-SIMS.