Decline in platelet microparticles contributes to reduced hemostatic potential of stored plasma.

Author(s): Matijevic N, Wang YW, Kostousov V, Wade CE, Vijayan KV, Holcomb JB

Publication: Thromb Res, 2011, Vol. 128, Page 35-41

PubMed ID: 21421259 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of refrigerated storage duration, filtration of plasma and thrombin generation reagents on MPs, residual cell counts, thrombin generation, and clot formation in thawed plasma. Blood was collected in citrate-phosphate-dextrose (CPD), and plasma was prepared by centrifugation and frozen until analysis. Frozen plasma was thawed at 37 degrees C, and, an aliquot was removed, analyzed immediately for residual cell counts, and frozen at -80 degrees C for other assays.

   Summary of Findings:

   The amount of platelet MPs was strongly correlated with the residual platelet count (r=0.75, p<0.001). Most of the platelet MPs bound annexin V. After 5 days of refrigeration, total MP counts in plasma decreased by 48% (p<0.0001) and platelet MPs decreased by 50% (p<0.0001) compared to levels measured in plasma immediately after thawing, while leukocyte, red blood cell, monocyte, and endothelial MPs decreased non-significantly. Further, the procoagulant MP activity decreased by 29% during refrigeration of plasma (p<0.0001), and procoagulant activity was modestly correlated with MP count (R=0.69, p<0.001). The peak thrombin generation and ETP were highly variable between specimens, and peak thrombin, ETP and rate decreased, while time to peak thrombin and lag time increased significantly, regardless of reagent used, when specimens were refrigerated for 5 days after thawing instead of analyzed immediately. The authors report peak thrombin and rate of thrombin generation were identified as discriminators of freshly thawed and aged plasma. TEG showed increased time to MRTG, time to MA, reaction time and SP in specimens refrigerated for 5 days after thawing compared to those analyzed immediately (p<0.001, p<0.01, p<0.0001, and p<0.0001, respectively), but alpha angle, MA, and MRTG were not affected by storage of thawed plasma. Filtration of plasma significantly reduced platelet MP count by 95% (p<0.001), procoaglulant activity by 36% (p<0.0001), MA (p<0.05), maximum elastic module (p<0.05), total thrombus generation (p<0.05) peak thrombin (p<0.01), ETP (p<0.01), and increased SP (p<0.05) and lag time (p<0.01). When MPs were added back to filtered thawed plasma, thrombogram parameters were comparable to those observed in unfiltered thawed plasma.

   Biospecimens

   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry
   Morphology	Hematology/ auto analyzer
   Protein	ELISA

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0 days 5 days
   Biospecimen Aliquots and Components	Filtration	Unfiltered Filtered
   Hematology/ auto analyzer Specific	Reaction solution	PRP reagent MP reagent PPP-low reagent

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