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Blood, Vol. 94 No. 6 (September 15), 1999:
pp. 1926-1932
Rapid Differentiation of a Rare Subset of Adult Human
Lin CD34CD38 Cells
Stimulated by Multiple Growth Factors In Vitro
Tomoaki Fujisaki,
Marc G. Berger,
Stefan Rose-John, and
Connie J. Eaves
From the Terry Fox Laboratory, British Columbia Cancer Agency, and
the University of British Columbia, Vancouver, British Columbia,
Canada; and I. Medizinische Klinik, Johannes Gutenberg Universitat
Mainz, Mainz, Germany.
Recently, several reports of lineage-negative (lin )
CD34 cells with in vivo hematopoietic activity have
focused interest on the properties and growth factor response
characteristics of these cells. We have now identified a combination of
5 growth factors that are necessary and sufficient to stimulate a
marked mitogenic and differentiation response by a subset of human
linCD34CD38 cells present
in normal adult human marrow and granulocyte colony-stimulating factor
(G-CSF)-mobilized blood. Less than 0.1% of the cells in highly
purified (including doubly sorted)
linCD34CD38 cells from
these 2 sources formed colonies directly in semisolid medium or
generated such cells after 6 weeks in long-term culture. Nevertheless,
approximately 1% of the same
linCD34CD38 cells were
able to proliferate rapidly in serum-free liquid suspension cultures
containing human flt-3 ligand, Steel factor, thrombopoietin, interleukin-3 (IL-3), and hyper-IL-6 to produce a net 28- ± 8-fold increase in total cells within 10 days. Of the cells present in these
10-day cultures, 5% ± 2% were CD34+ and 2.5% ± 0.9% were erythroid, granulopoietic, megakaryocytopoietic, or
multilineage colony-forming cells (CFC) (13 ± 7 CFC per
linCD34CD38 pre-CFC). In
contrast to linCD34+CD38
cells, this response of
linCD34CD38 cells required
exposure to all of the 5 growth factors used. Up to 1.7 × 105 linCD34 adult marrow
cells failed to engraft sublethally irradiated
NOD/SCID- 2M/ mice. These studies
demonstrate unique properties of a rare subset of
linCD34CD38 cells present
in both adult human marrow and mobilized blood samples that allow their
rapid proliferation and differentiation in vitro within an overall
period of 3 to 4 weeks. The rapidity of this response challenges
current concepts about the normal duration and coordinated control of
these processes in adults.
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