Published online before print August 5, 2004, doi: 10.1161/01.RES.0000141700.96085.2e
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© 2004 American Heart Association, Inc.
UltraRapid Communications |
Generation of Functional Culture–Derived Platelets From CD34+ Progenitor Cells to Study Transgenes in the Platelet Environment
From 1. Medizinische Klinik, Klinikum rechts der Isar und Deutsches Herzzentrum, Technische Universität München (S.M., C.S., M.G.); ProCorde (M.U., M.P., A.G., G.M.), Martinsried; and GSF-Forschungszentrum für Strahlen- und Umweltforschung, Institut für Pathologie (U.H.), Munich, Germany.
Correspondence to Prof Dr M. Gawaz, 1. Medizinische Klinik, Klinikum rechts der Isar und Deutsches Herzzentrum, Technische Universität München, Ismaningerstrasse 22, 81675 München. E-mail meinrad.gawaz{at}med1.med.tu-muenchen.de; or PD Dr M. Ungerer, ProCorde GmbH, Fraunhoferstrasse 9, 82152 Martinsried, Germany. E-mail ungerer@procorde.com
Abstract:— The possibility of evaluating the function of transgenes in platelets requires the generation of platelets from nucleated progenitor cells in vitro. In this article, we provide effective culture conditions for generating functional culture-derived (CD) human and mouse platelets from CD34+ progenitor cells that allow expression of any foreign protein of interest. We have evolved an effective cytokine cocktail (thrombopoietin, stem cell factor, interleukin [IL]-1ß, IL-6) that induces a high yield of CD platelets and optimal shedding from cultivated megakaryocytes generated from CD34+ progenitor cells. CD platelets showed similar functional and morphological characteristics compared with isolated blood platelets, including surface expression of platelet antigens (CD41, CD42, CD62P), aggregation, release of granule constituents (P-selectin, platelet factor 4, serotonin). Moreover, transmission electron microscopy revealed the presence of typical 
- and dense granules and dense tubular system in CD platelets. Additionally, we showed that stable transgene expression in CD platelets can be performed through infection of CD34+ progenitor cells using adenoviral vectors. Thus, we describe a methodology that enables studying functional consequences of transgenes of interest in the natural environment of platelets that may impose substantial impact on potential future platelet research and therapeutic target evaluation. The full text of this article is available online at http://circres.ahajournals.org.
Key Words: platelets • progenitor cells • megakaryopoiesis • thrombosis
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