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Growth and differentiation of a human megakaryoblastic cell line, CMK
N Komatsu, T Suda, M Moroi, N Tokuyama, Y Sakata, M Okada, T Nishida, Y Hirai, T Sato and A Fuse
Department of Medicine and Biochemistry II, Jichi Medical School, Tochigi,
Japan.
Recently, a human megakaryoblastic cell line, CMK, was established from the
peripheral blood of a megakaryoblastic leukemia patient with Down syndrome.
Using this cell line, we studied the proliferation and differentiation of
megakaryocytic cells in the presence of highly purified human hematopoietic
factors and phorbol 12-myristate-13- acetate (PMA). In a methylcellulose
culture system, interleukin-3 (IL- 3) and granulocyte-macrophage
colony-stimulating factor (GM-CSF) facilitated colony formation by CMK
cells in a dose-dependent manner. The maximum stimulating doses of these
factors were 10 and 200 U/mL, respectively. These concentrations were
comparable to those that stimulate activity in normal hematopoietic cells.
In contrast, granulocyte-colony stimulating factor (G-CSF),
macrophage-colony stimulating factor (M-CSF), and erythropoietin (EPO) had
no effects on the colony formation of CMK cells. In a liquid culture
system, 20% of the CMK cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa)
antigen without hematopoietic factors, whereas 40% of the cells expressed
GPIIb/IIIa with the addition of IL-3 and GM-CSF. EPO also slightly enhanced
expression of GPIIb/IIIa. On the other hand, PMA inhibited growth of CMK
cells and induced most of them to express the GPIIb/IIIa antigen.
Furthermore, PMA induced CMK cells to produce growth activity toward new
inocula of CMK cells. This growth factor (GF) contained colony-stimulating
activity (CSA) in normal bone marrow (BM) cells. The activity was believed
to be attributable mainly to GM-CSF, since 64% of this activity was
neutralized by anti-GM-CSF antibodies and a transcript of GM-CSF was
detected in mRNA from PMA-treated CMK cells by Northern blot analysis.
These observations suggest that GM-CSF, as well as IL-3, should play an
important role in megakaryocytopoiesis.
Volume 74,
Issue 1,
pp. 42-48,
07/01/1989
Copyright © 1989 by The American Society of Hematology
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