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Interactions between recombinant human erythropoietin and serum factor(s)
on murine megakaryocyte colony formation
J Tsukada, M Misago, M Kikuchi, T Sato, R Ogawa, T Ota, S Oda, I Morimoto, S Chiba and S Eto
First Department of Internal Medicine, School of Medicine, University of
Occupational and Environmental Health, Fukuoka, Japan.
We investigated the interactions between human erythropoietin (hEpo) and
serum factor(s) on murine megakaryocyte (MK) colony formation. Serum-free
cultures supported the growth of a large number of murine MK colonies in
the presence of murine interleukin-3 (mIL-3). The addition of fetal calf
serum (FCS) to mIL-3-containing cultures resulted in only a minimal
increase in the number of murine MK colonies. In contrast, hEpo alone had
no murine MK colony-stimulating activities in serum-free cultures. hEpo
required the presence of FCS, murine serum, or human serum in cultures to
promote murine MK colony growth and synergized with these sera to stimulate
murine MK colony formation. Furthermore, sera from patients with aplastic
anemia showed higher synergistic activities with hEpo than sera from
hematologically normal persons (normal human serum). When normal human
serum was fractionated by gel- filtration chromatography, two peaks with
the synergistic activity were observed in the eluent. However, serum did
not show any synergistic effects with hEpo on the growth of murine GM
colonies or murine colony- forming unit-erythroid-derived colonies.
Although human serum synergized with hEpo to stimulate murine MK colony
formation, human cytokines such as IL-3, IL-4, IL-6, granulocyte-macrophage
colony- stimulating factor (GM-CSF) and granulocyte-CSF (G-CSF) failed to
induce murine MK colony formation in Epo-containing cultures. In cultures
containing human IL-1 alpha + human IL-6 + hEpo as well as in cultures
containing hEpo, human IL-3 and human GM-CSF failed to show stimulatory
effects on murine MK colony formation. Moreover, the synergistic activity
of human serum with hEpo could not be neutralized by antibodies such as
antihuman IL-1 alpha, antihuman IL-3, antihuman IL-4, antihuman IL-6,
antihuman G-CSF, and antihuman GM-CSF. Our data show that serum contains a
growth factor(s) that synergizes with Epo to stimulate the proliferation
and differentiation of MK precursors, and strongly suggest that this
factor(s) is an unique growth factor(s) that is distinct from IL-1 alpha,
IL-3, IL-4, IL-6, G-CSF, and GM-CSF.
Volume 80,
Issue 1,
pp. 37-45,
07/01/1992
Copyright © 1992 by The American Society of Hematology
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