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Cryopreservation alters the membrane and cytoskeletal protein profile of platelet microparticles
Author(s) -
Raynel Sarah,
Padula Matthew P.,
Marks Denese C.,
Johnson Lacey
Publication year - 2015
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.13165
Subject(s) - cryopreservation , microparticle , gelsolin , platelet , flow cytometry , gpvi , chemistry , phosphatidylserine , microbiology and biotechnology , thrombin , andrology , biology , immunology , platelet activation , biochemistry , actin , membrane , phospholipid , embryo , medicine , astrobiology
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. STUDY DESIGN AND 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. CONCLUSION 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.