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Enhancement of murine blast cell colony formation in culture by recombinant
rat stem cell factor, ligand for c-kit
K Tsuji, KM Zsebo and M Ogawa
Department of Medicine, Medical University of South Carolina.
Mice with W mutations characterized by hypopigmentation, sterility, anemia,
and mast cell deficiency have abnormalities in c-kit, a receptor with
tyrosine kinase activity. Recently, the ligand for c-kit was cloned by
investigators in several laboratories. Zsebo et al identified and cloned a
gene for a cytokine termed stem cell factor (SCF) in the medium conditioned
by buffalo rat liver cells, and this cytokine proved to be c-kit ligand. We
have examined the effects of recombinant rat SCF (rrSCF) on colony
formation from primitive hematopoietic progenitors in culture. rrSCF and
erythropoietin (Ep) supported formation of granulocyte/macrophage (GM)
colonies as well as a small number of multilineage and blast cell colonies
from marrow cells of normal mice. We then examined the effects of rrSCF
using marrow and spleen cells of mice that had been treated with 150 mg/kg
5- fluorouracil (5-FU). Unlike single factors, combinations of factors such
as rrSCF plus interleukin-3 (IL-3), rrSCF plus IL-6, and rrSCF plus
granulocyte colony-stimulating factor (G-CSF) markedly stimulated the
growth of multilineage colonies. In contrast to these factor combinations
and a combination of IL-3 and IL-6, a combination of rrSCF and IL-4 did not
support multilineage colony formation. Mapping studies of the development
of multipotential blast cell colonies further indicated that rrSCF, like
IL-6, G-CSF, and IL-11, shortens the dormant period in which the stem cells
reside. When we tested the effects of rrSCF using pooled blast cells, which
are highly enriched for progenitors and are devoid of stromal cells, rrSCF
plus Ep supported formation of only a few multilineage colonies, indicating
that rrSCF itself is ineffective in support of the proliferation of
multipotential progenitors. However, rrSCF supported formation of a
significant number of neutrophil and neutrophil/macrophage colonies from
pooled blast cells, indicating that rrSCF is able to support directly the
proliferation of progenitors in neutrophil/monocyte lineages. c-kit ligand
may play important roles in adult hematopoiesis.
Volume 78,
Issue 5,
pp. 1223-1229,
09/01/1991
Copyright © 1991 by The American Society of Hematology
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