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Long-term generation of human mast cells in serum-free cultures of CD34+
cord blood cells stimulated with stem cell factor and interleukin- 3
B Durand, G Migliaccio, NS Yee, K Eddleman, T Huima-Byron, AR Migliaccio and JW Adamson
Laboratory of Hematopoietic Growth Factors, New York Blood Center, New York
10021.
The generation of murine mast cells is supported by several cytokines, and
mast cell lines are frequently established in long-term cultures of normal
murine marrow cells. In contrast, growth of human mast cells was initially
dependent on coculture with murine fibroblasts. The growth factor produced
by murine fibroblasts and required to observe differentiation of human mast
cells is attributable in part to stem cell factor (SCF). However, other
factors are likely involved. We have previously shown that the combination
of SCF and interleukin-3 (IL-3) efficiently sustains proliferation and
differentiation of colony- forming cells (CFCs) from pre-CFC enriched from
human umbilical cord blood by CD34+ selection. With periodic medium changes
and the addition of fresh growth factors, five consecutive cultures of
different cord blood samples gave rise to differentiated cells and CFCs for
more than 2 months. Although differentiated cells continued to be generated
for more than 5 months, CFCs were no longer detectable by day 50 of
culture. The cells have the morphology of immature mast cells, are
Toluidine blue positive, are karyotypically normal, are CD33+, CD34-,
CD45+, c-kit-, and c-fms-, and die in the absence of either SCF or IL- 3.
These cells do not form colonies in semisolid culture and are propagated in
liquid culture stimulated with SCF and IL-3 at a seeding concentration of
no less than 10(4) cells/mL. At refeedings, the cultures contain a high
number (> 50%) of dead cells and have a doubling time ranging from 5 to
12 days. This suggests that subsets of the cell population die because of a
requirement for a growth factor other than SCF or IL-3. These results
indicate that the combination of cord blood progenitor and stem cells, plus
a cocktail of growth factors including SCF and IL-3, is capable with high
efficiency of giving rise in serum-deprived culture to human mast cells
that behave like factor- dependent cell lines. These cells may represent a
useful tool for studies of human mast cell differentiation and leukemia.
Volume 84,
Issue 11,
pp. 3667-3674,
12/01/1994
Copyright © 1994 by The American Society of Hematology
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