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A comparative study of the phenotype and proliferative capacity of
peripheral blood (PB) CD34+ cells mobilized by four different protocols and
those of steady-phase PB and bone marrow CD34+ cells
LB To, DN Haylock, T Dowse, PJ Simmons, S Trimboli, LK Ashman and CA Juttner
Leukaemia Research Unit, Hanson Centre for Cancer Research, Institute of
Medical and Veterinary Science, Adelaide, Australia.
Peripheral blood (PB) CD34+ cells from four commonly used mobilization
protocols were studied to compare their phenotype and proliferative
capacity with steady-state PB or bone marrow (BM) CD34+ cells. Mobilized PB
CD34+ cells were collected during hematopoietic recovery after
myelosuppressive chemotherapy with or without granulocyte- macrophage
colony-stimulating factor (GM-CSF) or granulocyte colony- stimulating
factor (G-CSF) or during G-CSF administration alone. The expression of
activation and lineage-associated markers and c-kit gene product were
studied by flow cytometry. Proliferative capacity was measured by
generation of nascent myeloid progenitor cells (granulocyte- macrophage
colony-stimulating factor; CFU-GM) and nucleated cells in a stroma-free
liquid culture stimulated by a combination of six hematopoietic growth
factors (interleukin-1 (IL-1), IL-3, IL-6, GM-CSF, G-CSF, and stem cell
factor). G-CSF-mobilized CD34+ cells have the highest percentage of CD38-
cells (P < .0081), but otherwise, CD34+ cells from different
mobilization protocols were similar to one another in their phenotype and
proliferative capacity. The spectrum of primitive and mature myeloid
progenitors in mobilized PB CD34+ cells was similar to their steady-state
counterparts, but the percentages of CD34+ cells expressing CD10 or CD19
were lower (P < .0028). Although steady-state PB and
chemotherapy-mobilized CD34+ cells generated fewer CFU-GM at day 21 than
G-CSF-mobilized and steady-state BM CD34+ cells (P < .0449), the
generation of nucleated cells and CFU-GM were otherwise comparable. The
presence of increased or comparable numbers of hematopoietic progenitors
within PB collections with equivalent proliferative capacity to BM CD34+
cells is not unexpected given the rapid and complete hematopoietic
reconstitution observed with mobilized PB. However, all four types of
mobilized PB CD34+ cells are different from steady-state BM CD34+ cells in
that they express less c-kit (P < .0002) and CD71 (P < .04) and
retain less rhodamine 123 (P < .0001). These observations are novel and
suggest that different mobilization protocols may act via similar pathways
involving the down-regulation of c-kit and may be independent of cell-cycle
status.
Volume 84,
Issue 9,
pp. 2930-2939,
11/01/1994
Copyright © 1994 by The American Society of Hematology
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