|
|
 Previous Article | Table of Contents | Next Article 
Two-color flow cytometric measurement of DNA distributions of rat
megakaryocytes in unfixed, unfractionated marrow cell suspensions
CW Jackson, LK Brown, BC Somerville, SA Lyles and AT Look
The ploidy distribution of megakaryocytes shifts in response to platelet
demand and thus provides a sensitive index of megakaryocytopoiesis. Flow
cytometry (FCM) is a potentially valuable method for rapid determination of
ploidy distributions of megakaryocyte populations; however, because
megakaryocytes constitute only a very small proportion of the cells in
unfractionated marrow, other rare events, such as cell clumping, complicate
FCM analysis. We describe the measurement of cellular DNA distributions of
megakaryocytes by two- color FCM in unfixed, unfractionated marrow--a
method based on the resistance of megakaryocytes to hypotonic lysis in the
cold for at least 2 days. Specific platelet antiserum was used to label
megakaryocytes by indirect immunofluorescence with fluorescein (green
fluorescence), and DNA was stained with propidium iodide (red fluorescence)
in hypotonic citrate solution. The ploidy distribution of megakaryocytes
was selectively determined with two-color, green-gated FCM, with which the
red and green fluorescence of all cells is analyzed, but only the red
fluorescence (DNA content) of cells that specifically bound the platelet
antibody is recorded. We demonstrate that this method can readily detect
changes in megakaryocyte DNA distributions due to experimental
thrombocytopenia or platelet hypertransfusion and, therefore, should be
useful for both experimental and clinical investigations of
megakaryocytopoiesis.
Volume 63,
Issue 4,
pp. 768-778,
04/01/1984
Copyright © 1984 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
H. Wang, S. Cnhen, Y. Chen, H. Wang, H. Wu, H. Tang, W. Xiong, J. Ma, Y. Ge, Q. Lu, et al.
The role of Tyro 3 subfamily receptors in the regulation of hemostasis and megakaryocytopoiesis
Haematologica,
May 1, 2007;
92(5):
643 - 650.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Tiedt, T. Schomber, H. Hao-Shen, and R. C. Skoda
Pf4-Cre transgenic mice allow the generation of lineage-restricted gene knockouts for studying megakaryocyte and platelet function in vivo
Blood,
February 15, 2007;
109(4):
1503 - 1506.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. S. Alexander, E. M. Viney, J.-G. Zhang, D. Metcalf, M. Kauppi, C. D. Hyland, M. R. Carpinelli, W. Stevenson, B. A. Croker, A. A. Hilton, et al.
Thrombocytopenia and kidney disease in mice with a mutation in the C1galt1 gene
PNAS,
October 31, 2006;
103(44):
16442 - 16447.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. P. McCormack, M. A. Hall, S. M. Schoenwaelder, Q. Zhao, S. Ellis, J. A. Prentice, A. J. Clarke, N. J. Slater, J. M. Salmon, S. P. Jackson, et al.
A critical role for the transcription factor Scl in platelet production during stress thrombopoiesis
Blood,
October 1, 2006;
108(7):
2248 - 2256.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Wernig, T. Mercher, R. Okabe, R. L. Levine, B. H. Lee, and D. G. Gilliland
Expression of Jak2V617F causes a polycythemia vera-like disease with associated myelofibrosis in a murine bone marrow transplant model
Blood,
June 1, 2006;
107(11):
4274 - 4281.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. B. Slayton, D. A. Wainman, X. M. Li, Z. Hu, A. Jotwani, C. R. Cogle, D. Walker, R. C. Fisher, J. R. Wingard, E. W. Scott, et al.
Developmental Differences in Megakaryocyte Maturation Are Determined by the Microenvironment
Stem Cells,
September 1, 2005;
23(9):
1400 - 1408.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. D. Growney, H. Shigematsu, Z. Li, B. H. Lee, J. Adelsperger, R. Rowan, D. P. Curley, J. L. Kutok, K. Akashi, I. R. Williams, et al.
Loss of Runx1 perturbs adult hematopoiesis and is associated with a myeloproliferative phenotype
Blood,
July 15, 2005;
106(2):
494 - 504.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. L. Masters, K. R. Palmer, W. S. Stevenson, D. Metcalf, E. M. Viney, N. S. Sprigg, W. S. Alexander, N. A. Nicola, and S. E. Nicholson
Genetic Deletion of Murine SPRY Domain-Containing SOCS Box Protein 2 (SSB-2) Results in Very Mild Thrombocytopenia
Mol. Cell. Biol.,
July 1, 2005;
25(13):
5639 - 5647.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Metcalf, M. R. Carpinelli, C. Hyland, S. Mifsud, L. DiRago, N. A. Nicola, D. J. Hilton, and W. S. Alexander
Anomalous megakaryocytopoiesis in mice with mutations in the c-Myb gene
Blood,
May 1, 2005;
105(9):
3480 - 3487.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Toki, F. Katsuoka, R. Kanezaki, G. Xu, H. Kurotaki, J. Sun, T. Kamio, S. Watanabe, S. Tandai, K. Terui, et al.
Transgenic expression of BACH1 transcription factor results in megakaryocytic impairment
Blood,
April 15, 2005;
105(8):
3100 - 3108.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Tomer
Human marrow megakaryocyte differentiation: multiparameter correlative analysis identifies von Willebrand factor as a sensitive and distinctive marker for early (2N and 4N) megakaryocytes
Blood,
November 1, 2004;
104(9):
2722 - 2727.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. Tong and H. F. Lodish
Lnk Inhibits Tpo-mpl Signaling and Tpo-mediated Megakaryocytopoiesis
J. Exp. Med.,
September 7, 2004;
200(5):
569 - 580.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. E. Geddis and K. Kaushansky
Megakaryocytes express functional Aurora-B kinase in endomitosis
Blood,
August 15, 2004;
104(4):
1017 - 1024.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Chang, P. A. Nakagawa, S. A. Williams, M. R. Schwartz, K. L. Imfeld, J. S. Buzby, and D. J. Nugent
Immune thrombocytopenic purpura (ITP) plasma and purified ITP monoclonal autoantibodies inhibit megakaryocytopoiesis in vitro
Blood,
August 1, 2003;
102(3):
887 - 895.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Kaluzhny, G. Yu, S. Sun, P. A. Toselli, B. Nieswandt, C. W. Jackson, and K. Ravid
BclxL overexpression in megakaryocytes leads to impaired platelet fragmentation
Blood,
August 13, 2002;
100(5):
1670 - 1678.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Angchaisuksiri, S. R. Grigus, P. L. Carlson, G. W. Krystal, and E. N. Dessypris
Secretion of a unique peptide from interleukin-2-stimulated natural killer cells that induces endomitosis in immature human megakaryocytes
Blood,
January 1, 2002;
99(1):
130 - 136.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S.-M. Luoh, E. Stefanich, G. Solar, H. Steinmetz, T. Lipari, T. I. Pestina, C. W. Jackson, and F. J. de Sauvage
Role of the Distal Half of the c-Mpl Intracellular Domain in Control of Platelet Production by Thrombopoietin In Vivo
Mol. Cell. Biol.,
January 15, 2000;
20(2):
507 - 515.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
J. Lin and D. I. H. Linzer
Induction of Megakaryocyte Differentiation by a Novel Pregnancy-specific Hormone
J. Biol. Chem.,
July 23, 1999;
274(30):
21485 - 21489.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Vyas, K. Ault, C. W. Jackson, S. H. Orkin, and R. A. Shivdasani
Consequences of GATA-1 Deficiency in Megakaryocytes and Platelets
Blood,
May 1, 1999;
93(9):
2867 - 2875.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Vitrat, K. Cohen-Solal, C. Pique, J. P. LeCouedic, F. Norol, A. K. Larsen, A. Katz, W. Vainchenker, and N. Debili
Endomitosis of Human Megakaryocytes Are Due to Abortive Mitosis
Blood,
May 15, 1998;
91(10):
3711 - 3723.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Norol, N. Vitrat, E. Cramer, J. Guichard, S. A. Burstein, W. Vainchenker, and N. Debili
Effects of Cytokines on Platelet Production From Blood and Marrow CD34+ Cells
Blood,
February 1, 1998;
91(3):
830 - 843.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Ishida, T. Ito, and S.-i. Kuriya
Effects of C-Mpl Ligand on Cytoplasmic Maturation of Murine Megakaryocytes and on Platelet Production
J. Histochem. Cytochem.,
January 1, 1998;
46(1):
49 - 58.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
J. T. Arnold, N. C. Daw, P. E. Stenberg, D. Jayawardene, D. K. Srivastava, and C. W. Jackson
A Single Injection of Pegylated Murine Megakaryocyte Growth and Development Factor (MGDF ) Into Mice Is Sufficient to Produce a Profound Stimulation of Megakaryocyte Frequency, Size, and Ploidization
Blood,
February 1, 1997;
89(3):
823 - 833.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Thompson, Y. Zhang, D. Kamen, C. W. Jackson, R. D. Cardiff, and K. Ravid
Deregulated Expression of c-myc in Megakaryocytes of Transgenic Mice Increases Megakaryopoiesis and Decreases Polyploidization
J. Biol. Chem.,
September 20, 1996;
271(38):
22976 - 22982.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. J. Kuter
Thrombopoietin: Biology and Clinical Applications
Oncologist,
February 1, 1996;
1(1):
98 - 106.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Kimura, A. Hart, M. Hirashima, C. Wang, D. Holmyard, J. Pittman, X.-L. Pang, C. W. Jackson, and A. Bernstein
Zinc Finger Protein, Hzf, Is Required for Megakaryocyte Development and Hemostasis
J. Exp. Med.,
April 1, 2002;
195(7):
941 - 952.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
