Journal of Cell Science, Vol 97, Issue 1 59-70, Copyright © 1990 by Company of Biologists

JOURNAL ARTICLES

Blood platelet formation in vitro. The role of the cytoskeleton in megakaryocyte fragmentation

F Tablin, M Castro and RM Leven
Department of Anatomy, School of Veterinary Medicine, University of California, Davis 95616.

We have developed a unique in vitro model that promotes differentiation of megakaryocytes into platelets. When megakaryocytes isolated from guinea pig bone marrow were cultured on hydrated rat tail collagen gels, cells spontaneously formed elongated, beaded processes that fragmented to yield cytoplasmic pieces with the same size and internal composition as individual platelets. Addition of nocodazole at the initiation of cultures blocked process formation, while addition of nocodazole to cells with previously established processes resulted in their retraction. The addition of taxol to cultures resulted in abnormally thick processes that were tightly adherent to the underlying substratum, and did not bead or fragment. Cytochalasin D accelerated process formation and fragmentation of megakaryocytes cultured on collagen gels by twofold. On the basis of these results, we propose a model for platelet formation in culture that involves the following steps: adherence of megakaryocytes to the underlying extracellular matrix; dilation of the demarcation membrane system and breakdown of the actin-rich peripheral zone; microtubule-based extension of pseudopodia, which are no longer adherent to the substratum; and fragmentation into platelets by the coalescence and fusion of demarcation membrane vesicles with the plasma membrane. We feel that this distinctive culture system closely approximates thrombocytopoiesis in vivo, thus allowing detailed elucidation of this important process.

This article has been cited by other articles:

				M. K. Larson and S. P. Watson Regulation of proplatelet formation and platelet release by integrin {alpha}IIbbeta3 Blood, September 1, 2006; 108(5): 1509 - 1514. [Abstract] [Full Text] [PDF]

				H. Schulze, M. Korpal, J. Hurov, S.-W. Kim, J. Zhang, L. C. Cantley, T. Graf, and R. A. Shivdasani Characterization of the megakaryocyte demarcation membrane system and its role in thrombopoiesis Blood, May 15, 2006; 107(10): 3868 - 3875. [Abstract] [Full Text] [PDF]

				C. S. Abrams Packing platelets to go Blood, December 15, 2005; 106(13): 4019 - 4020. [Full Text] [PDF]

				J. L. Richardson, R. A. Shivdasani, C. Boers, J. H. Hartwig, and J. E. Italiano Jr Mechanisms of organelle transport and capture along proplatelets during platelet production Blood, December 15, 2005; 106(13): 4066 - 4075. [Abstract] [Full Text] [PDF]

				S. R. Patel, J. L. Richardson, H. Schulze, E. Kahle, N. Galjart, K. Drabek, R. A. Shivdasani, J. H. Hartwig, and J. E. Italiano Jr Differential roles of microtubule assembly and sliding in proplatelet formation by megakaryocytes Blood, December 15, 2005; 106(13): 4076 - 4085. [Abstract] [Full Text] [PDF]

				H. Schulze, M. Korpal, W. Bergmeier, J. E. Italiano Jr, S. M. Wahl, and R. A. Shivdasani Interactions between the megakaryocyte/platelet-specific {beta}1 tubulin and the secretory leukocyte protease inhibitor SLPI suggest a role for regulated proteolysis in platelet functions Blood, December 15, 2004; 104(13): 3949 - 3957. [Abstract] [Full Text] [PDF]

				H. Kojima, H. Kanada, S. Shimizu, E. Kasama, K. Shibuya, H. Nakauchi, T. Nagasawa, and A. Shibuya CD226 Mediates Platelet and Megakaryocytic Cell Adhesion to Vascular Endothelial Cells J. Biol. Chem., September 19, 2003; 278(38): 36748 - 36753. [Abstract] [Full Text] [PDF]

				S. de Botton, S. Sabri, E. Daugas, Y. Zermati, J. E. Guidotti, O. Hermine, G. Kroemer, W. Vainchenker, and N. Debili Platelet formation is the consequence of caspase activation within megakaryocytes Blood, July 30, 2002; 100(4): 1310 - 1317. [Abstract] [Full Text] [PDF]

				R. Zunino, Q. Li, S. D. Rose, M. M. I. Romero-Benitez, T. Lejen, N. C. Brandan, and J.-M. Trifaro Expression of scinderin in megakaryoblastic leukemia cells induces differentiation, maturation, and apoptosis with release of plateletlike particles and inhibits proliferation and tumorigenesis Blood, October 1, 2001; 98(7): 2210 - 2219. [Abstract] [Full Text] [PDF]

				R. A. Shivdasani Molecular and Transcriptional Regulation of Megakaryocyte Differentiation Stem Cells, September 1, 2001; 19(5): 397 - 407. [Abstract] [Full Text] [PDF]

				P. Rojnuckarin and K. Kaushansky Actin reorganization and proplatelet formation in murine megakaryocytes: the role of protein kinase C{alpha} Blood, January 1, 2001; 97(1): 154 - 161. [Abstract] [Full Text] [PDF]

				P. Lecine, J. E. Italiano Jr, S.-W. Kim, J.-L. Villeval, and R. A. Shivdasani Hematopoietic-specific beta 1 tubulin participates in a pathway of platelet biogenesis dependent on the transcription factor NF-E2 Blood, August 15, 2000; 96(4): 1366 - 1373. [Abstract] [Full Text] [PDF]

				J. E. Italiano , Jr., P. Lecine, R. A. Shivdasani, and J. H. Hartwig Blood Platelets Are Assembled Principally at the Ends of Proplatelet Processes Produced by Differentiated Megakaryocytes J. Cell Biol., December 13, 1999; 147(6): 1299 - 1312. [Abstract] [Full Text] [PDF]

				P. Lecine, J.-L. Villeval, P. Vyas, B. Swencki, Y. Xu, and R. A. Shivdasani Mice Lacking Transcription Factor NF-E2 Provide In Vivo Validation of the Proplatelet Model of Thrombocytopoiesis and Show a Platelet Production Defect That Is Intrinsic to Megakaryocytes Blood, September 1, 1998; 92(5): 1608 - 1616. [Abstract] [Full Text] [PDF]

				G. Zauli, M. Vitale, E. Falcieri, D. Gibellini, A. Bassini, C. Celeghini, M. Columbaro, and S. Capitani In Vitro Senescence and Apoptotic Cell Death of Human Megakaryocytes Blood, September 15, 1997; 90(6): 2234 - 2243. [Abstract] [Full Text] [PDF]

				E. M. Cramer, F. Norol, J. Guichard, J. Breton-Gorius, W. Vainchenker, J.-M. Masse, and N. Debili Ultrastructure of Platelet Formation by Human Megakaryocytes Cultured With the Mpl Ligand Blood, April 1, 1997; 89(7): 2336 - 2346. [Abstract] [Full Text] [PDF]