Blood online
Home About Blood Authors Subscriptions Permission Advertising Public Access contact us
 

 
Advanced
Current Issue
First Edition
Future Articles
Archives
Submit to Blood
Search
American Society of Hematology
Meeting Abstracts
Email Alerts
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Right arrow Rights and Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Faraday, N.
Right arrow Articles by Bray, P. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Faraday, N.
Right arrow Articles by Bray, P. F.
Related Collections
Right arrow Hematopoiesis and Stem Cells
Social Bookmarking
 Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

arrow to previous article Previous Article  |  Table of Contents  |  Next Article next article arrow

Blood, Vol. 94 No. 12 (December 15), 1999: pp. 4084-4092

Ex Vivo Cultured Megakaryocytes Express Functional Glycoprotein IIb-IIIa Receptors and Are Capable of Adenovirus-Mediated Transgene Expression

Nauder Faraday, Jeffrey J. Rade, David C. Johns, Gopal Khetawat, Stephen J. Noga, John F. DiPersio, Ying Jin, Janet L. Nichol, Jeff S. Haug, and Paul F. Bray

From the Department of Anesthesiology and Critical Care Medicine, the Divisions of Cardiology and Hematology, the Department of Medicine, and the Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD; the Division of BMT and Cell Biology, Washington University School of Medicine, St Louis, MO; and Amgen, Inc, Thousand Oaks, CA.

Investigation of the molecular basis of megakaryocyte (MK) and platelet biology has been limited by an inadequate source of genetically manipulable cells exhibiting physiologic MK and platelet functions. We hypothesized that ex vivo cultured MKs would exhibit agonist inducible glycoprotein (GP) IIb-IIIa activation characteristic of blood platelets and that these cultured MKs would be capable of transgene expression. Microscopic and flow cytometric analyses confirmed that human hematopoietic stem cells cultured in the presence of pegylated recombinant human MK growth and development factor (PEG-rHuMGDF) differentiated into morphologic and phenotypic MKs over 2 weeks. Cultured MKs expressed functional GPIIb-IIIa receptors as assessed by agonist inducible soluble fibrinogen and PAC1 binding. The specificity and kinetics of fibrinogen binding to MK GPIIb-IIIa receptors were similar to those described for blood platelets. The reversibility and internalization of ligands bound to MK GPIIb-IIIa also shared similarities with those observed in platelets. Cultured MKs were transduced with an adenoviral vector encoding green fluorescence protein (GFP) or beta -galactosidase (beta -gal). Efficiency of gene transfer increased with increasing multiplicities of infection and incubation time, with 45% of MKs expressing GFP 72 hours after viral infection. Transduced MKs remained capable of agonist induced GPIIb-IIIa activation. Thus, ex vivo cultured MKs (1) express agonist responsive GPIIb-IIIa receptors, (2) are capable of expressing transgenes, and (3) may prove useful for investigation of the molecular basis of MK differentiation and GPIIb-IIIa function.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
Arterioscler. Thromb. Vasc. Bio.Home page
A. Gillitzer, M. Peluso, K.-L. Laugwitz, G. Munch, S. Massberg, I. Konrad, M. Gawaz, and M. Ungerer
Retroviral Infection and Selection of Culture-Derived Platelets Allows Study of the Effect of Transgenes on Platelet Physiology Ex Vivo and on Thrombus Formation In Vivo
Arterioscler Thromb Vasc Biol, August 1, 2005; 25(8): 1750 - 1755.
[Abstract] [Full Text] [PDF]


Home page
Circ. Res.Home page
M. Ungerer, M. Peluso, A. Gillitzer, S. Massberg, U. Heinzmann, C. Schulz, G. Munch, and M. Gawaz
Generation of Functional Culture-Derived Platelets From CD34+ Progenitor Cells to Study Transgenes in the Platelet Environment
Circ. Res., September 3, 2004; 95(5): e36 - e44.
[Abstract] [Full Text] [PDF]


Home page
Proc. Natl. Acad. Sci. USAHome page
K. Eto, R. Murphy, S. W. Kerrigan, A. Bertoni, H. Stuhlmann, T. Nakano, A. D. Leavitt, and S. J. Shattil
Megakaryocytes derived from embryonic stem cells implicate CalDAG-GEFI in integrin signaling
PNAS, October 1, 2002; 99(20): 12819 - 12824.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
A. Bertoni, S. Tadokoro, K. Eto, N. Pampori, L. V. Parise, G. C. White, and S. J. Shattil
Relationships between Rap1b, Affinity Modulation of Integrin alpha IIbbeta 3, and the Actin Cytoskeleton
J. Biol. Chem., July 5, 2002; 277(28): 25715 - 25721.
[Abstract] [Full Text] [PDF]


Home page
BloodHome page
M. Majka, A. Janowska-Wieczorek, J. Ratajczak, M. A. Kowalska, G. Vilaire, Z. K. Pan, M. Honczarenko, L. A. Marquez, M. Poncz, and M. Z. Ratajczak
Stromal-derived factor 1 and thrombopoietin regulate distinct aspects of human megakaryopoiesis
Blood, December 15, 2000; 96(13): 4142 - 4151.
[Abstract] [Full Text] [PDF]



 click for free articles
home about blood authors subscriptions permissions advertising public access contact us
  Copyright © 1999 by American Society of Hematology         Online ISSN: 1528-0020