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Maintenance of high levels of pluripotent hematopoietic stem cells in
vitro: effect of stromal cells and c-kit
JP Wineman, S Nishikawa and CE Muller-Sieburg
Medical Biology Institute, La Jolla, CA 92037.
We show here that mouse pluripotent hematopoietic stem cells can be
maintained in vitro on stroma for at least 3 weeks at levels close to those
found in bone marrow. The extent of stem cell maintenance is affected by
the nature of the stromal cells. The stromal cell line S17 supported stem
cells significantly better than heterogeneous, primary stromal layers or
the stromal cell line Strofl-1. Stem cells cultured on S17 repopulated all
hematopoietic lineages in marrow-ablated hosts for at least 10 months,
indicating that this culture system maintained primitive stem cells with
extensive proliferative capacity. Furthermore, we demonstrate that, while
pluripotent stem cells express c-kit, this receptor appears to play only a
minor role in stem cell maintenance in vitro. The addition of an antibody
that blocks the interaction of c-kit with its ligand essentially abrogated
myelopoiesis in cultures. However, the level of stem cells in
antibody-treated cultures was similar to that found in untreated cultures.
Thus, it seems likely that the maintenance of primitive stem cells in vitro
depends on yet unidentified stromal cell-derived factor(s).
Volume 81,
Issue 2,
pp. 365-372,
01/15/1993
Copyright © 1993 by The American Society of Hematology
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