title: Cryopreservation alters the membrane and cytoskeletal protein profile of platelet microparticles.
abstract: BACKGROUND
  Cryopreservation of platelets (PLTs) in dimethyl sulfoxide (DMSO) and storage at -80 °C extends the PLT shelf life to at least 2 years, allowing greater accessibility in military and rural environments. While cryopreserved PLTs have been extensively characterized, the microparticles formed as a result of cryopreservation are yet to be fully described.
  
  
  METHODS
  Apheresis PLTs were cryopreserved at -80 °C with 5% DMSO and sampled before freezing and after thawing. Microparticle number, size, surface receptor phenotype, and function were assessed by microscopy, flow cytometry, dynamic light scattering, and thrombin-generating capacity. Proteomic changes were examined using two-dimensional gel electrophoresis and Western blotting.
  
  
  RESULTS
  PLT cryopreservation resulted in a 15-fold increase in the number of microparticles compared to fresh PLTs. The surface receptor phenotype of these microparticles differed to microparticles from fresh PLTs, with more microparticles expressing glycoprotein (GP)IV, GPIIb, and the GPIb-V-IX complex. Cryopreservation drastically altered the abundance of many cytoskeletal proteins in the PLT microparticles, including actin, filamin, gelsolin, and tropomyosin. Despite these changes, PLT microparticles were functional and contributed to phosphatidylserine- and tissue factor- induced thrombin generation.
  
  
  CONCLUSIONS
  This study demonstrates that PLT microparticles formed by cryopreservation are phenotypically distinct from those present before freezing. These differences may be associated with the procoagulant properties of cryopreserved PLTs.
authors.affiliation: Research & Development, Australian Red Cross Blood Service, Alexandria, NSW, Australia.|Proteomics Core Facility, University of Technology Sydney, Sydney, NSW, Australia.
authors.forename: Sarah
authors.initials: S
authors.lastname: Raynel
authors.affiliation: Proteomics Core Facility, University of Technology Sydney, Sydney, NSW, Australia.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Padula
authors.affiliation: Research & Development, Australian Red Cross Blood Service, Alexandria, NSW, Australia.
authors.forename: Denese C
authors.initials: DC
authors.lastname: Marks
authors.affiliation: Research & Development, Australian Red Cross Blood Service, Alexandria, NSW, Australia.
authors.forename: Lacey
authors.initials: L
authors.lastname: Johnson
mesh_terms: D001792:Blood Platelets / Q000378:metabolism*; D001793:Blood Preservation; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D015925:Cryopreservation; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D005260:Female; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D040901:Proteomics / Q000379:methods
chemical_list: D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D004121:Dimethyl Sulfoxide