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12.08.2016 | Original Article | Ausgabe 11/2016

Annals of Hematology 11/2016

A whole blood model of thrombocytopenia that controls platelet count and hematocrit

Annals of Hematology > Ausgabe 11/2016
R. S. Bercovitz, M. K. Brenner, D. K. Newman
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1007/​s00277-016-2777-9) contains supplementary material, which is available to authorized users.


In patients with thrombocytopenia, it can be difficult to predict a patient’s bleeding risk based on platelet count alone. Platelet reactivity may provide additional information; however, current clinical assays cannot reliably assess platelet function in the setting of thrombocytopenia. New methods to study platelet reactivity in thrombocytopenic samples are needed. In this study, we sought to develop a laboratory model of thrombocytopenia using blood from healthy subjects that preserves the whole blood environment and reproducibly produces samples with a specific platelet count and hematocrit. We compared the activation state of unstimulated and agonist-stimulated platelets in thrombocytopenic samples derived from this method with normocytic controls. Whole blood was diluted with autologous red blood cell concentrate and platelet-poor plasma, which were obtained via centrifugation, in specific ratios to attain a final sample with a predetermined platelet count and hematocrit. P-selectin exposure and GPIIbIIIa activation in unstimulated platelets and platelets stimulated with collagen-related peptide (CRP) or adenosine diphosphate (ADP) in thrombocytopenic samples and the normocytic control from which they were derived were quantified by flow cytometry. Our methodology reliably produced thrombocytopenic samples with a platelet count ≤50,000/μL and an accurately and precisely controlled hematocrit. P-selectin exposure and GPIIbIIIa activation on unstimulated platelets or on ADP- or CRP-stimulated platelets did not differ in thrombocytopenic samples compared to normocytic controls. We describe a new method for creating thrombocytopenic blood that can be used to better understand the contributions of platelet number and function to hemostasis.

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Supplemental Material 1 Description of how the equations to calculate volumes of whole blood, red cell concentrate, and platelet-poor plasma should be added together to get the reconstituted whole blood with the goal platelet count and hematocrit. (PDF 1675 kb)
Supplemental Material 2 Calculator to determine volumes of whole blood, RBCs, and PPP to use to achieve goal hematocrit and platelet count. (XLS 21 kb)
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