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Possible mechanisms accounting for the growth factor independence of
hematopoietic progenitors from umbilical cord blood
KR Schibler, Y Li, RK Ohls, NC Nye, MC Durham, W White, KW Liechty, T Le and RD Christensen
Division of Human Development and Aging, University of Utah School of
Medicine, Salt Lake City 84132.
Hematopoietic progenitors obtained from the bone marrow of healthy adults
fail to undergo clonogenic maturation in vitro if a source of hematopoietic
growth factors is not included in the culture dishes. In contrast, a
fraction of similarly purified progenitors obtained from umbilical cord
blood undergo clonogenic maturation even in the absence of added growth
factors. We postulated that production of hematopoietic growth factors
within the culture dishes containing the progenitors of umbilical cord
blood origin might be responsible. We postulated further, that this
production might be by non-progenitor cells co- plated along with the
progenitors, or alternatively by CD34+ cells themselves, or by cells
clonally derived from CD34+ cells. To test these possibilities we first
assessed the effect of including in the cultures neutralizing antibody
directed against various growth factors. Inclusion of anti-granulocyte
macrophage colony-stimulating factor (GM- CSF) and anti-interleukin-3
(IL-3) (but not anti-IL-2) significantly reduced the growth factor
independence of cord blood progenitors (P < .005 and P < .01).
Inclusion of both anti-GM-CSF and anti-IL-3 almost completely ablated the
spontaneous colony growth (P < .001). Inclusion of IL-10 also reduced,
in a concentration-dependent fashion, the spontaneous generation of
umbilical cord blood-derived colonies. Transcripts for GM-CSF and IL-3 were
detected, by reverse transcriptase- polymerase chain reaction (RT-PCR), in
the CD34+ cells from cord blood and from adult marrow. When plated without
added growth factors, however, the CD34+ cells of adult marrow origin
failed to produce colonies, whereas 6% of cord blood CD34+ cells similarly
cultured did so. When these growth factor independent colonies were plucked
from culture, transcripts for GM-CSF and IL-3 were identified in all. We
conclude that production of GM-CSF and IL-3 occurs within culture dishes
containing hematopoietic progenitors of umbilical cord origin, and that
this explains some of their apparently unique features of in vitro growth.
Volume 84,
Issue 11,
pp. 3679-3684,
12/01/1994
Copyright © 1994 by The American Society of Hematology
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