Lipidomic and proteomic characterization of platelet extracellular vesicle subfractions from senescent platelets

Transfusion. 2015 Mar;55(3):507-21. doi: 10.1111/trf.12874. Epub 2014 Oct 21.


Background: Platelets (PLTs) in stored PLT concentrates (PLCs) release PLT extracellular vesicles (PL-EVs) induced by senescence and activation, resembling the PLT storage lesion. No comprehensive classification or molecular characterization of senescence-induced PL-EVs exists to understand PL-EV heterogeneity.

Study design and methods: PL-EVs from 5-day-stored PLCs from healthy individuals were isolated and subfractionated by differential centrifugation, filtration, and density gradient ultracentrifugation into five PLT microvesicle (PL-MV) subfractions (Fraction [F]1-F5) and PLT exosomes (PL-EXs). PL-EV size, concentration, and composition were analyzed by nanoparticle tracking analysis, flow cytometry, and lipid and protein mass spectrometry. Protein data were verified by Western blot.

Results: PL-EVs showed overlapping mean particle sizes of 180 to 260 nm, but differed significantly in composition. Less dense, intermediate, and dense PL-MVs enriched specific lipidomic and proteomic markers related to the plasma membrane, intracellular membranes, PLT granules, mitochondria, and PLT activation. α-Synuclein (81% of total) accumulated in F1 and F2, amyloid-β (Aβ) precursor protein in F3 and F4 (84%), and apolipoprotein (Apo)E (88%) and ApoJ (92%) in F3 to F5. PL-EXs enriched lipid species and proteins, with high abundance of lipid raft, PLT adhesion, and immune response-related markers.

Conclusion: Differential lipid and protein compositions of PL-EVs suggest their unique cellular origins and functions, partly overlapping with PLT granule secretion. Dense PL-MVs might represent autophagic vesicles released during PLT activation and apoptosis and PL-EXs resemble lipid rafts, with a potential role in PLT aggregation and immunity. Segregation of α-synuclein and Aβ precursor protein, ApoE, and ApoJ into less dense and dense PL-MVs, respectively, show their differential carrier role of neurologic disease-related cargo.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Blood Platelets / chemistry*
  • Blood Platelets / cytology
  • Blood Preservation
  • Blood Proteins / isolation & purification*
  • Blotting, Western
  • Cell-Derived Microparticles / chemistry*
  • Cellular Senescence
  • Centrifugation / methods
  • Erythrocytes / chemistry
  • Exosomes / chemistry*
  • Filtration
  • Flow Cytometry
  • Humans
  • Immunomagnetic Separation
  • Lipids / blood*
  • Lipids / isolation & purification
  • Mass Spectrometry / methods
  • Membrane Lipids / blood
  • Membrane Lipids / isolation & purification
  • Membrane Proteins / blood
  • Membrane Proteins / isolation & purification
  • Nanoparticles
  • Nerve Degeneration
  • Platelet Activation
  • Plateletpheresis
  • alpha-Synuclein / blood


  • Blood Proteins
  • Lipids
  • Membrane Lipids
  • Membrane Proteins
  • alpha-Synuclein