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Recombinant rat stem cell factor stimulates the amplification and
differentiation of fractionated mouse stem cell populations
N Williams, I Bertoncello, H Kavnoudias, K Zsebo and I McNiece
Department of Physiology, University of Melbourne, Parkville, Victoria,
Australia.
The role of recombinant rat stem cell factor (rrSCF) was studied on defined
primitive bone marrow cell populations. In agar culture of 500
lineage-negative/Sca-1-positive (Lin-/Sca-1+) cells, rrSCF alone stimulates
small colonies of predominantly granulocytic cells. The combinations of
rrSCF plus interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating
factor (GM-CSF), or macrophage CSF (CSF-1) stimulated primitive progenitor
cells defined as high proliferative potential colony-forming cells
(HPP-CFC). Synergistic increases in total colony numbers were obtained with
rrSCF plus GM-CSF, granulocyte CSF (G-CSF), CSF-1, or IL-6, but not IL-1 or
IL-3. Lin-/Sca-1+ cells were incubated in liquid culture at 3,000 cells/mL
for 6 days in the presence of rrSCF alone or in combination with other
growth factors. The total number of cells was increased twofold in the
presence of rrSCF, with the progeny primarily myeloid in nature. The
greatest increase in cell number was obtained with rrSCF plus IL-3, where
the cell number increased 40-fold. These factors also stimulated an
increase in HPP-CFC (10-fold) and GM-CFC (500-fold). To determine if these
interactions were direct, single Lin-/Sca-1+ cells were sorted into
microtiter wells and the cell proliferation scored 6 days later. RrSCF
synergized with IL-3, IL-6, and G-CSF to stimulate the proliferation of
single cells. The cells in positive wells were subcultured into
colony-forming assays and up to 400 CFC per well were obtained after 14
days incubation of the secondary cultures. These data demonstrate that
rrSCF acts in combination with various growth factors to directly stimulate
the amplification potential of hematopoietic primitive precursors,
resulting in differentiation of these precursors.
Volume 79,
Issue 1,
pp. 58-64,
01/01/1992
Copyright © 1992 by The American Society of Hematology
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