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Platelets. 2019;30(4):428-437. doi: 10.1080/09537104.2018.1513473. Epub 2018 Oct 4.

Flow cytometry for pediatric platelets.

Author information

1
a Cellular Hemostasis and Thrombosis Lab , National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare , Moscow , Russian Federation.
2
b Faculty of Biology, Moscow State University named after M.V. Lomonosov , Moscow , Russian Federation.
3
c Day Hospital , National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare , Moscow , Russian Federation.
4
d Neonatal Intensive Care and Resuscitation Unit , National Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov, Russian Ministry of Healthcare , Moscow , Russian Federation.
5
e Hematological Center , City Clinical Hospital named after S.P. Botkin , Moscow , Russia.
6
f Institute of Hematology, Immunology and Cell Technologies , National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare , Moscow , Russian Federation.
7
g Medical administration , National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare , Moscow , Russian Federation.
8
h Faculty of Biological and Medical Physics , Moscow Institute of Physics and Technology , Dolgoprudny , Russian Federation.
9
i Laboratory of Molecular Mechanisms of Hemostasis , Center for Theoretical Problems of Physicochemical Pharmacology , Moscow , Russian Federation.

Abstract

The ability of platelets to carry out their hemostatic function can be impaired in a wide range of inherited and acquired conditions: trauma, surgery, inflammation, pre-term birth, sepsis, hematological malignancies, solid tumors, chemotherapy, autoimmune disorders, and many others. Evaluation of this impairment is vitally important for research and clinical purposes. This problem is particularly pronounced in pediatric patients, where these conditions occur frequently, while blood volume and the choice of blood collection methods could be limited. Here we describe a simple flow cytometry-based screening method of comprehensive whole blood platelet function testing that was validated for a range of pediatric and adult samples (n = 31) in the hematology hospital setting including but not limited to: classic inherited platelet function disorders (Glanzmann's thrombasthenia; Bernard-Soulier, Wiscott-Aldrich, and Hermasky-Pudlak syndromes, MYH9-dependent thrombocytopenia), healthy and pre-term newborns, acute and chronic immune thrombocytopenia, chronic lympholeukemia, effects of therapy on platelet function, etc. The method output includes levels of forward and side scatter, levels of major adhesion and aggregation glycoproteins Ib and IIb-IIIa, active integrins' level based on PAC-1 binding, major alpha-granule component P-selectin, dense granule function based on mepacrine uptake and release, and procoagulant activity quantified as a percentage of annexin V-positive platelets. This analysis is performed for both resting and dual-agonist-stimulated platelets. Preanalytical and analytical variables are provided and discussed. Parameter distribution within the healthy donor population for adults (n = 72) and children (n = 17) is analyzed.

KEYWORDS:

Flow cytometry; immune thrombocytopenia; pediatric platelets; platelet function; thrombocytopathy

PMID:
30285517
DOI:
10.1080/09537104.2018.1513473
[Indexed for MEDLINE]

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