Faculty Collaboration Database

A whole blood model of thrombocytopenia that controls platelet count and hematocrit. Ann Hematol 2016 Oct;95(11):1887-94

Date

08/16/2016

Pubmed ID

27515424

Pubmed Central ID

PMC6055520

DOI

10.1007/s00277-016-2777-9

Scopus ID

2-s2.0-84981501486 (requires institutional sign-in at Scopus site)   9 Citations

Abstract

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.

Author List

Bercovitz RS, Brenner MK, Newman DK

Authors

Michelle Brenner in the CTSI department at Medical College of Wisconsin - CTSI
Debra K. Newman PhD Investigator in the Blood Research Institute department at BloodCenter of Wisconsin
Debra K. Newman PhD Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Adenosine Diphosphate
Adult
Carrier Proteins
Centrifugation
Flow Cytometry
Hematocrit
Hemorrhagic Disorders
Humans
In Vitro Techniques
P-Selectin
Peptides
Platelet Activation
Platelet Count
Platelet Function Tests
Platelet Glycoprotein GPIIb-IIIa Complex
Reproducibility of Results
Thrombocytopenia