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Flow cytometric analysis of megakaryocytes from patients with abnormal
platelet counts
A Tomer, P Friese, R Conklin, W Bales, L Archer, LA Harker and SA Burstein
Department of Basic and Clinical Research, Scripps Clinic, La Jolla, CA.
Megakaryocytes (MKs) from 40 patients with quantitative platelet disorders
and 19 normal volunteers were analyzed by flow cytometry for size, fine
cell internal structure and granularity, membrane expression of the
glycoprotein (GP) IIb/IIIa complex, and for ploidy distribution. Analysis
was performed on unfractionated minimally manipulated marrows obtained from
routine bone marrow aspirates. MKs were labeled with a fluorescent
lineage-specific monoclonal antibody to the GPIIb/IIIa complex followed by
DNA staining with propidium iodide. Eight hundred to 3,000 MKs were
analyzed in each sample. The modal ploidy distribution in normals was 16N,
comprising about half of the megakaryocytic population, with 22.6% of the
cells less than or equal to 8N and 22.0% greater than or equal to 32N.
Twelve thrombocytopenic patients with decreased marrow MKs on biopsy (mean
platelet count [MPC] 44,600/microliters) showed an increase in low ploidy
cells with 53.2% less than or equal to 8N (P less than .01); cell size was
reduced in three patients when compared to normal cells of identical ploidy
(P less than .05). Eight thrombocytopenic patients with enhanced platelet
destruction (with normal or increased MKs on biopsy and shortened platelet
survival; MPC 41,400/microliters) showed an increased proportion of high
ploidy cells greater than or equal to 32N to 39.2% (P less than .01).
Increased cell size and granularity were found in four of these patients (P
less than .05). Six patients with thrombocytopenia secondary to multiple
mechanisms affecting both platelet production and destruction (MPC
66,700/microliters) showed no shift in ploidy. Four patients with primary
thrombocytosis (two with thrombocythemia and two with polycythemia vera;
MPC 822,500/microliters) showed a marked shift toward high ploidy cells
with 42.3% greater than or equal to 32N and 7.6% greater than or equal to
64N cells (P less than .01). The shift was accompanied by a marked increase
in cell size and granularity in the patients with thrombocythemia. Ten
patients with thrombocytosis secondary to chronic blood loss, malignant or
inflammatory disorders (MPC 714,000/microliters), showed variable
distributions with four patients exhibiting a shift in ploidy to the right
similar to that found in the patients with increased platelet destruction.
Based upon the present data, flow cytometric ploidy distribution may be
diagnostically useful in thrombocytopenic patients by discriminating
between disorders of platelet production and destruction. (ABSTRACT
TRUNCATED AT 400 WORDS)
Volume 74,
Issue 2,
pp. 594-601,
08/01/1989
Copyright © 1989 by The American Society of Hematology
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