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Platelet concentrates prepared and stored under currently optimal conditions: minor impact on platelet adhesive and cohesive functions after storage
Authors:M. Lozano  E. Estebanell  J. Cid  M. Diaz-Ricart  R. Mazzara  A. Ordinas   G. Escolar
Affiliation:Department of Hemotherapy, August Pi i Sunyer Institute for Biomedical Investigations, Hospital Clínic, University of Barcelona, Spain. mlozano@medicina.ub.es
Abstract:BACKGROUND: The effect on platelets of two standard methods of platelet concentrate (PC) preparation was studied by flow cytometry. The findings were correlated with those obtained in an experimental in vitro perfusion model. STUDY DESIGN AND METHODS: PCs were prepared from whole blood by the platelet-rich plasma (PRP) or buffy coat (BC) method and placed on a flatbed platelet agitator at 22 degrees C for up to 5 days. Platelet glycoproteins (GP)lbalpha, GPIIb/IIIa, and GPIV, p-selectin and lysosomal integral membrane protein, and the binding of von Willebrand factor, fibrinogen, fibronectin, and coagulation factor Va were measured with the corresponding specific conjugated antibodies. Perfusions were carried out in an annular chamber with citrated blood depleted of platelets and white cells by filtration, to which samples from PCs were added. RESULTS: PRP-PC production provoked intense platelet activation. In contrast, in BC-derived PCs, platelet activation was milder, and only a significant increase in bound fibrinogen was seen. After 1 day of storage, differences between the methods that had been observed immediately after separation had almost disappeared. During the remaining storage period, increases in activation-dependent antigens and in procoagulant activity were measured. Of the studied platelet GPs, only GPIIIb/ IIIa decreased by 25 percent in PRP-PCs. Differences in covered surface were not significant in perfusion studies performed on Day 0 and after 5 days of storage in PRP-PCs (26.8 +/- 6.9 vs. 20.5 +/- 5.8) or BC-PCs (23.8 +/- 11 vs. 24.8 +/- 10.2). CONCLUSION: Platelet activation occurred during the separation and storage of PCs prepared by both methods, and it was higher in PRP-PCs only in samples obtained immediately after preparation. Despite these changes, platelet adhesive and cohesive functions were similar in both types of PCs and remained basically unchanged after storage.
Keywords:BC = buffy coat    FITC = fluorescein isothiocyanate    GP(s) = glycoprotein(s)    LIMP = lysosomal integral membrane protein    MFI = mean fluorescence intensity    MoAb(s) = monoclonal antibody(ies)    PC(s) = platelet concentrate(s)    PE = phycoerythrin    PRP = platelet-rich plasma    vWF = von Willebrand factor
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