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Long-term generation of colony-forming cells in liquid culture of CD34+
cord blood cells in the presence of recombinant human stem cell factor
G Migliaccio, AR Migliaccio, ML Druzin, PJ Giardina, KM Zsebo and JW Adamson
Laboratory of Hematopoietic Growth Factors, Lindsley F. Kimball Research
Institute, New York Blood Center, New York 10021.
Human cord blood was used as a source of progenitor and stem cells to
evaluate the effect of recombinant human stem-cell factor (SCF) on colony
formation and the generation of colony-forming cells (CFC) under highly
defined, serum-deprived conditions. SCF interacted with a number of
hematopoietic growth factors to stimulate colony growth and was
particularly effective in stimulating the formation of mixed-cell colonies
from CD34+ soybean agglutinin negative (SBA-) cells. In suspension culture
of CD34+, SBA- cells, SCF alone was unable to maintain cell numbers or CFC
but, in combination with interleukin-3 (IL- 3), increased input numbers of
cells by 10-fold and increased CFC of all kinds by nearly 20-fold. This
included erythroid burst-forming cells (BFU-E), granulocyte/macrophage (GM)
CFC, and mixed-cell CFC. In contrast, CD34- SBA- cells neither gave rise to
CFC nor were maintained by combinations of growth factors including SCF.
SCF interacted with erythropoietin (Epo) and granulocyte colony-stimulating
factor (G-CSF) to maintain large numbers of cells as well as to generate a
twofold to threefold increase in CFC in the case of Epo, and a 10-fold
increase in CFC in the case of G-CSF. With Epo, the predominant CFC
generated were BFU-E and erythroid CFC and many of the cells in suspension
were erythroblasts. In contrast, SCF plus G-CSF resulted in large numbers
of granulocytes at various stages of maturation and the CFC generated were
almost exclusively granulocytic-CFC. IL-1 and IL-6, alone or in combination
with SCF, showed little or no ability to increase cell numbers or generate
CFC. In summary, SCF interacts with a variety of hematopoietic growth
factors to promote colony formation, particularly mixed-cell colony
formation, and also, in suspension culture, SCF interacts with IL-3, G-CSF,
and Epo to generate large numbers of differentiated cells as well as a
variety of CFC for up to 1 month.
Volume 79,
Issue 10,
pp. 2620-2627,
05/15/1992
Copyright © 1992 by The American Society of Hematology
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