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Blood, 1 August 2002, Vol. 100, No. 3, pp. 869-878
HEMATOPOIESIS
Functional analysis of human hematopoietic repopulating cells
mobilized with granulocyte colony-stimulating factor alone versus
granulocyte colony-stimulating factor in combination with stem
cell factor
David A. Hess,
Krysta D. Levac,
Francis N. Karanu,
Michael Rosu-Myles,
Martin J. White,
Lisa Gallacher,
Barbara Murdoch,
Michael Keeney,
Pamela Ottowski,
Ronan Foley,
Ian Chin-Yee, and
Mickie Bhatia
From Developmental Stem Cell Biology and Regenerative
Medicine, John P. Robarts Research Institute; the Department of
Microbiology and Immunology, University of Western Ontario; and the
London Health Sciences Center, London, Ontario; and Hamilton Health
Sciences Center, Ontario.
Using in vitro progenitor assays, serum-free in vitro cultures, and
the nonobese diabetic/severe combined immune-deficient (NOD/SCID)
ecotropic murine virus knockout xenotransplantation model to
detect human SCID repopulating cells (SRCs) with multilineage reconstituting function, we have characterized and compared purified subpopulations harvested from the peripheral blood (PB) of patients receiving granulocyte colony-stimulating factor (G-CSF) alone or in
combination with stem cell factor (SCF). Mobilized G-CSF plus
SCF PB showed a 2-fold increase in total mononuclear cell content and a
5-fold increase in CD34-expressing cells depleted for lineage-marker
expression (CD34+Lin ) as compared with
patients treated with G-CSF alone. Functionally, G-CSF plus
SCF-mobilized CD34+CD38Lin
cells contained a 2-fold enhancement in progenitor frequency as
compared with G-CSF-mobilized subsets. Despite enhanced cellularity and progenitor capacity, G-CSF plus SCF mobilization did not
increase the frequency of SRCs as determined by limiting dilution
analysis by means of unfractionated PB cells. Purification of SRCs from these sources demonstrated that as few as 1000 CD34+CD38Lin cells from
G-CSF-mobilized PB contained SRC capacity while G-CSF plus
SCF-mobilized CD34+CD38Lin
cells failed to repopulate at doses up to 500 000 cells. In addition, primitive
CD34CD38AC133+Lin
cells derived from G-CSF plus SCF-mobilized PB were capable of differentiation into CD34-expressing cells, while the identical subfractions from G-CSF PB were unable to produce CD34+
cells in serum-free cultures. Our study defines qualitative and quantitative distinctions among subsets of primitive cells mobilized by
means of G-CSF plus SCF versus G-CSF alone, and therefore has implications for the utility of purified repopulating cells from these sources.
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