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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
1   Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France
,
Elisabeth M. Cramer
1   Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France
,
Gulie Alimardani
1   Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France
,
Serge Fichelson
1   Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France
,
Jean-Marc Massé
1   Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France
,
Catherine P. M. Hayward
1   Departments of Pathology and Molecular Medicine and Medicine, McMaster University, Hamilton, Ontario, Canada INSERM U474, Hôpital de Port Royal, Paris, France
› Author AffiliationsFinancial support: Supported by grant 42450 from the Canadian Institutes of Health Research (C.P.M.H.) and a Premier’s Research Excellence Award from the Government of Ontario. C.P.M.H. is the recipient of a Career Investigator Award from the Heart and Stroke Foundation of Ontario and a Canada Research Chair in Molecular Hemostasis.
Further Information

Publication History

Received 28 February 2003

Accepted after resubmission 27 July 2003

Publication Date:
05 December 2017 (online)

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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
 

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