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Granulocyte-macrophage colony-stimulating factor-dependent growth and
erythropoietin-induced differentiation of a human cell line MB-02
DA Morgan, DL Gumucio and I Brodsky
Department of Neoplastic Diseases, Hahnemann University, Philadelphia, PA
19102.
Peripheral blood blasts from a patient with acute megakaryoblastic leukemia
were placed into liquid cultures with recombinant growth factors. Growth,
but not differentiation, was supported by interleukin- 3 (IL-3) or
granulocyte-macrophage colony-stimulating factor (GM-CSF) for the first 30
days of culture. Sustained growth occurred only with GM-CSF and gave rise
to the cell line MB-02, which has been in continuous culture for over 1
year. The cell line retained the surface phenotype of the leukemic
megakaryoblasts except for the loss of glycoproteins Ib and IIb/IIIa, which
were induced after exposure to phorbol esters. The induction of
erythropoiesis occurred when GM-CSF- deprived cells were cultured with
erythropoietin (Epo). Well-defined morphologic stages of differentiation
ranging from primitive erythroblasts to nuclei-extruding normoblasts were
seen. Transforming growth factor-beta inhibited GM-CSF- and Epo-dependent
growth, but not erythroid maturation. Indirect immunofluorescence using
globin chain- specific monoclonal antibodies detected fetal, but not adult
hemoglobin in the uninduced cells. beta-globin was induced and gamma-globin
was increased after Epo exposure. Both globin species accumulated in the
developing erythrocytes until terminal differentiation. Quantitative S1
analysis of beta-like globin transcripts showed very low levels of epsilon-
and beta-globin expression and high levels of gamma-globin expression in
cells maintained in GM-CSF. Five days after induction with Epo, epsilon
message decreased to barely detectable levels while gamma and beta
transcripts increased threefold and 20-fold, respectively. This novel cell
line not only retains many characteristics of the leukemic megakaryoblasts
from which it was derived, but also can be induced to recapitulate apparent
normal erythropoiesis.
Volume 78,
Issue 11,
pp. 2860-2871,
12/01/1991
Copyright © 1991 by The American Society of Hematology
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