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The unexpected G0/G1 cell cycle status of mobilized hematopoietic stem
cells from peripheral blood
N Uchida, D He, AM Friera, M Reitsma, D Sasaki, B Chen and A Tsukamoto
Systemix Inc, Palo Alto, CA 94304, USA.
Treatment with a combination of cytokines and chemotherapy can effectively
stimulate the release of hematopoietic stem cells (HSC) into the peripheral
blood (PB), which can then be harvested for transplantation. The cell cycle
status of the harvested HSC from mobilized PB (MPB) is of interest because
of the impact that cell cycling may have on optimizing the conditions for
ex vivo expansion, retrovirus-mediated gene transfer, and the engraftment
of transplanted tissues. Therefore, we characterized the cell cycling
status of mobilized HSC from mice and humans. The murine HSC, which express
the phenotype c-kit+ Thy-1.1lo Lin-/lo Sca-1+, were purified from PB, bone
marrow (BM), and spleen after the mice were treated with the mobilizing
regimen of granulocyte colony-stimulating factor (G-CSF) or a combination
of cyclophosphamide (CTX) and G-CSF. Human HSC (CD34+ Thy- 1+ Lin-) and
progenitor cells (CD34+ Thy-1-Lin-) were isolated from the BM of untreated
healthy volunteers and from MPB of healthy volunteers and patients treated
with G-CSF or a combination of CTX and GM-CSF. Cell cycle status was
determined by quantitating the amount of DNA in the purified cells after
staining with the dye Hoechst 33342. Fluorescence-activated cell sorting
analysis of the progenitor cells from the murine and human samples showed
an unexpected finding, ie, virtually none of the cells from the MPB was
cycling. The G0/G1 status of HSC from MPB was surprising, because a
significant proportion of HSC from BM are actively proliferating and, after
mobilization, the HSC in the spleen and BM were also actively cycling.
Volume 89,
Issue 2,
pp. 465-472,
01/15/1997
Copyright © 1997 by The American Society of Hematology
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