Studies of α-granule proteins in cultured human megakaryocytes

Dragoslava Kika Veljkovic; Elisabeth M. Cramer; Gulie Alimardani; Serge Fichelson; Jean-Marc Massé; Catherine P. M. Hayward

doi:10.1160/TH03-02-0125

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 90(05): 844-852
DOI: 10.1160/TH03-02-0125

Platelets and Blood Cells

Schattauer GmbH

Studies of α-granule proteins in cultured human megakaryocytes

Dragoslava Kika Veljkovic

¹Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France

,

Elisabeth M. Cramer

¹Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France

,

Gulie Alimardani

¹Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France

,

Serge Fichelson

¹Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France

,

Jean-Marc Massé

¹Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France

,

Catherine P. M. Hayward

¹Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France

› Author Affiliations

Abstract

Summary

α-Granule protein storage is important for producing platelets with normal haemostatic function. The low to undetectable levels of several megakaryocyte-synthesized α-granule proteins in normal plasma suggest megakaryocytes are important to sequester these proteins in vivo. α-Granule protein storage in vitrohas been studied using other cell types, with differences observed in how some proteins are processed compared to platelets. Human megakaryocytes, cultured from cord blood CD34⁺cells and grown in serum-free media containing thrombopoietin, were investigated to determine if they could be used as a model for studying normal α-granule protein processing and storage. ELISA indicated that cultured megakaryocytes contained the α-granule proteins multimerin, von Willebrand factor, thrombospondin-1, β-thromboglobulin and platelet factor 4, but no detectable fibrinogen and factor V. A significant proportion of the α-granule protein in megakaryocyte cultures was contained within the cells (averages: 41 – 71 %), consistent with storage. Detailed analyses of multimerin and von Willebrand factor confirmed that α-granule proteins were processed to mature forms and were predominantly located in the α-granules of cultured megakaryocytes. Thrombopoietin-stimulated cultured megakaryocytes provide a useful model for studying α-granule protein processing and storage.

Keywords

Megakaryocytes - platelets - α-granules - protein processing

Full Text

References

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