Original article

Blood Transfusion 2-2018 (March - April)

Application of a clot-based assay to measure the procoagulant activity of stored allogeneic red blood cell concentrates

Authors

Key words: extracellular vesicles, red blood cell concentrates, coagulation, storage
Publication Date: 2017-03-09

Abstract

Background. Thrombotic effects are possible complications of red blood cell transfusion. The generation and accumulation of procoagulant red blood cell extracellular vesicles during storage may play an important role in these thrombotic effects. The objective of this study was to assess the value of a simple phospholipid-dependent clot-based assay (STA®-Procoag-PPL) to estimate the procoagulant activity of stored red blood cells and changes in this activity during storage of the blood component.
Materials and methods. Extracellular vesicles from 12 red blood cell concentrates were isolated at 13 storage time-points and characterised by quantitative and functional methods: the degree of haemolysis (direct spectrophotometry), the quantification and determination of cellular origin (flow cytometry) and the procoagulant activity (thrombin generation and STA®-Procoag-PPL assays) were assessed.
Results. The mean clotting time of extracellular vesicles isolated from red blood cell concentrates decreased from 117.2±3.6 sec on the day of collection to 33.8±1.3 sec at the end of the storage period. This illustrates the phospholipid-dependent procoagulant activity of these extracellular vesicles, as confirmed by thrombin generation. Results of the peak of thrombin and the STA®-Procoag-PPL were well correlated (partial r=−0.41. p<0.001). In parallel, an exponential increase of the number of red blood cell-derived extracellular vesicles from 1,779/µL to 218,451/µL was observed.
Discussion. The STA®-Procoag-PPL is a potentially useful technique for assessing the procoagulant activity of a red blood cell concentrate.

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Authors

Bérangère Devalet - Department of Haematology, Université Catholique de Louvain, CHU UCL Namur, Namur Thrombosis and Hemostasis Center (NTHC), Yvoir

Adeline Wannez - Department of Pharmacy, University of Namur, Namur Thrombosis and Haemostasis Centre (NTHC), Namur

Nicolas Bailly - Haematology Laboratory, Université Catholique de Louvain, CHU UCL Namur, Namur Thrombosis and Hemostasis Center (NTHC), Yvoir

Lutfiye Alpan - 2Department of Pharmacy, University of Namur, Namur Thrombosis and Haemostasis Centre (NTHC), Namur

Damien v - Haematology Laboratory, Université Catholique de Louvain, CHU UCL Namur, Namur Thrombosis and Hemostasis Center (NTHC), Yvoir

Jonathan Douxfils - Department of Pharmacy, University of Namur, Namur Thrombosis and Haemostasis Centre (NTHC), Namur

Véronique Deneys - Blood Transfusion Centre-Mont-Godinne, Université Catholique de Louvain, CHU UCL Namur, Yvoir

Benoît Bihin - Scientific Support Unit, Université Catholique de Louvain, CHU UCL Namur, Yvoir, Belgium

Bernard Chatelain - Haematology Laboratory, Université Catholique de Louvain, CHU UCL Namur, Namur Thrombosis and Hemostasis Center (NTHC), Yvoir

Jean-Michel Dogné - Department of Pharmacy, University of Namur, Namur Thrombosis and Haemostasis Centre (NTHC), Namur

Christian Chatelain - Department of Pharmacy, University of Namur, Namur Thrombosis and Haemostasis Centre (NTHC), Namur

François Mullier - Haematology Laboratory, Université Catholique de Louvain, CHU UCL Namur, Namur Thrombosis and Hemostasis Center (NTHC), Yvoir

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