|
|
 Previous Article | Table of Contents | Next Article 
Flow cytometric analysis of normal human megakaryocytes
A Tomer, LA Harker and SA Burstein
Department of Basic and Clinical Research, Scripps Clinic and Research
Foundation, La Jolla, CA 92037.
Megakaryocytes from normal routine human bone marrow aspirates were
analyzed by flow cytometry for size, fine cell structure and granularity,
membrane expression of glycoprotein (GP) IIb/IIIa and ploidy. Marrow cells
were initially enriched for megakaryocytes by a Percoll density gradient
and megakaryocytes were labeled with a fluoresceinated monoclonal antibody
directed to the GPIIb/IIIa complex. The cells were fixed with
paraformaldehyde and stained with propidium iodide (PI) for DNA
quantitation. Using two-color flow cytometry, megakaryocytes were
identified by their high membrane immunofluorescence and their ploidy was
determined according to the relative fluorescence intensity of the PI.
Forward light scatter (FSC), correlating with cell size, 90 degrees side
light scatter (SSC), reflecting primarily cell internal fine structure and
granularity, and total cell membrane fluorescence were examined. To
evaluate independently the relationship between size and cell membrane
fluorescence obtained by flow cytometry, megakaryocytes were sorted
directly on slides and analyzed by a laser-based anchored cell analyzer
(ACAS). There was a strong correlation among size, SSC, and the level of
membrane fluorescence. The mean diameter of megakaryocytes was 28.1 +/-
12.3 micron. The modal ploidy distribution was 16N with approximately
one-fifth of the cells less than or equal to 4N. The mean FSC and SSC
levels increased with increasing ploidy. However, the marked overlap
observed between the ranges of these parameters in adjacent ploidy classes
suggested that size and SSC increase continuously rather than by discrete
steps as is characteristic for ploidy. The total surface membrane
fluorescence was correlated with cell size (r = 0.98) as measured by FSC or
directly by the ACAS (r = 0.85), and with cell ploidy (r = 0.99) indicating
an augmentation in total membrane GPIIb/IIIa expression with an increase in
cell size and ploidy. However, estimated GPIIb/IIIa fluorescence density
was inversely correlated with FSC suggesting that the GPIIb/IIIa surface
epitope density is decreased with increasing cell maturity. We conclude
that flow cytometry is a useful technique for the rapid analysis of human
megakaryocytes obtained by marrow aspiration, and should be applicable to
studies of pathologic states.
Volume 71,
Issue 5,
pp. 1244-1252,
05/01/1988
Copyright © 1988 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:
|
|
|
|
 
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]
|
|
|
|
|
|
|
A. Tomer
Effects of anagrelide on in vivo megakaryocyte proliferation and maturation in essential thrombocythemia
Blood,
March 1, 2002;
99(5):
1602 - 1609.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Shiotsu, K. Yamashita, F. Kanai, Y. Ikuina, C. Murakata, M. Teramura, H. Mizoguchi, T. Tamaoki, and S. Akinaga
Chemoprotective effects of KF41399, a derivative of carbazole compounds, on nimustine-induced thrombocytopenia
Blood,
June 15, 2000;
95(12):
3771 - 3780.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. A. Harker, L. K. Roskos, U. M. Marzec, R. A. Carter, J. K. Cherry, B. Sundell, E. N. Cheung, D. Terry, and W. Sheridan
Effects of megakaryocyte growth and development factor on platelet production, platelet life span, and platelet function in healthy human volunteers
Blood,
April 15, 2000;
95(8):
2514 - 2522.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Riviere, F. Subra, K. Cohen-Solal, V. Cordette-Lagarde, R. Letestu, C. Auclair, W. Vainchenker, and F. Louache
Phenotypic and Functional Evidence for the Expression of CXCR4 Receptor During Megakaryocytopoiesis
Blood,
March 1, 1999;
93(5):
1511 - 1523.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Stoffel, S. Ziegler, N. Ghilardi, B. Ledermann, F. J. de Sauvage, and R. C. Skoda
Permissive role of thrombopoietin and granulocyte colony-stimulating factor receptors in hematopoietic cell fate decisions in vivo
PNAS,
January 19, 1999;
96(2):
698 - 702.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. A. Harker, U. M. Marzec, F. Novembre, I. B. Sundell, E. K. Waller, S. Karpatkin, H. M. McClure, A. B. Kelly, and R. B. Stead
Treatment of Thrombocytopenia in Chimpanzees Infected With Human Immunodeficiency Virus by Pegylated Recombinant Human Megakaryocyte Growth and Development Factor
Blood,
June 15, 1998;
91(12):
4427 - 4433.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. L. Cole, U. M. Marzec, C. J. Gunthel, S. Karpatkin, L. Worford, I. B. Sundell, J. L. Lennox, J. L. Nichol, and L. A. Harker
Ineffective Platelet Production in Thrombocytopenic Human Immunodeficiency Virus-Infected Patients
Blood,
May 1, 1998;
91(9):
3239 - 3246.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. A. Harker, U. M. Marzec, A. B. Kelly, E. Cheung, A. Tomer, J. L. Nichol, S. R. Hanson, and R. B. Stead
Prevention of Thrombocytopenia and Neutropenia in a Nonhuman Primate Model of Marrow Suppressive Chemotherapy by Combining Pegylated Recombinant Human Megakaryocyte Growth and Development Factor and Recombinant Human Granulocyte Colony-Stimulating Factor
Blood,
January 1, 1997;
89(1):
155 - 165.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Ayala, A Tomer, and K. Kellar
Flow cytometric analysis of megakaryocyte-associated antigens on CD34 cells and their progeny in liquid culture
Stem Cells,
May 1, 1996;
14(3):
320 - 329.
[Abstract]
|
|
|
|
|
