Cryopreservation alters the membrane and cytoskeletal protein profile of platelet microparticles

Raynel, S; Padula, MP; Marks, DC; Johnson, L

Cryopreservation alters the membrane and cytoskeletal protein profile of platelet microparticles

Raynel, S Padula, MP

Marks, DC Johnson, L

Permalink

Publication Type:: Journal Article
Citation:: Transfusion, 2015, 55 (10), pp. 2422 - 2432
Issue Date:: 2015-10-01

Closed Access

	Filename	Description	Size
	Raynel_et_al-2015-Transfusion.pdf	Accepted Manuscript Version	525.37 kB	Adobe PDF	View/Open

Copyright Clearance Process

Recently Added
In Progress
Closed Access

This item is closed access and not available.

Full metadata record

Field	Value	Language
dc.contributor.author	Raynel, S	en_US
dc.contributor.author	Padula, MP https://orcid.org/0000-0002-8283-0643	en_US
dc.contributor.author	Marks, DC	en_US
dc.contributor.author	Johnson, L	en_US
dc.date.available	2015-04-02	en_US
dc.date.issued	2015-10-01	en_US
dc.identifier.citation	Transfusion, 2015, 55 (10), pp. 2422 - 2432	en_US
dc.identifier.issn	0041-1132	en_US
dc.identifier.uri	http://hdl.handle.net/10453/37330
dc.description.abstract	© 2015 AABB. 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.	en_US
dc.relation.ispartof	Transfusion	en_US
dc.relation.isbasedon	10.1111/trf.13165	en_US
dc.subject.classification	Cardiovascular System & Hematology	en_US
dc.subject.mesh	Blood Platelets	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Dimethyl Sulfoxide	en_US
dc.subject.mesh	Cytoskeletal Proteins	en_US
dc.subject.mesh	Membrane Proteins	en_US
dc.subject.mesh	Cryopreservation	en_US
dc.subject.mesh	Blood Preservation	en_US
dc.subject.mesh	Proteomics	en_US
dc.subject.mesh	Female	en_US
dc.subject.mesh	Male	en_US
dc.subject.mesh	Cell-Derived Microparticles	en_US
dc.title	Cryopreservation alters the membrane and cytoskeletal protein profile of platelet microparticles	en_US
dc.type	Journal Article
utslib.citation.volume	10	en_US
utslib.citation.volume	55	en_US
utslib.for	1103 Clinical Sciences	en_US
utslib.for	1107 Immunology	en_US
utslib.for	1102 Cardiorespiratory Medicine and Haematology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access
pubs.issue	10	en_US
pubs.publication-status	Published	en_US
pubs.volume	55	en_US

Abstract:

© 2015 AABB. 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.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/37330