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In vitro expansion of human peripheral blood CD34+ cells
N Sato, K Sawada, K Koizumi, T Tarumi, M Ieko, T Yasukouchi, M Yamaguchi, TA Takahashi, S Sekiguchi and T Koike
Department of Internal Medicine II, Hokkaido University School of Medicine,
Japan.
To elucidate the role of recombinant human colony-stimulating factors
(CSFs) for expanding peripheral blood (PB) CD34+ cells, these cells were
purified up to 94.5% +/- 1.3% and the effects of individual and combined
CSFs on the proliferation and differentiation of these cells were studied
in a 7-day suspension culture. The majority of CD34+ cells coexpressed CD38
(81.8% +/- 5.1%), but was negative for CD33 (88.5% +/- 3.4%). Among the
individual CSFs examined, recombinant interleukin-3 (rIL-3) was identified
as the most potent factor for expanding PB progenitor cells and increased
nonerythroid progenitor cells 13- +/- 4- fold (P < .01). Recombinant
granulocyte-macrophage colony-stimulating factor (rGM-CSF), recombinant
granulocyte-CSF (rG-CSF), recombinant macrophage-CSF (rM-CSF), rIL-6,
rIL-11, and recombinant stem cell factor (rSCF) did not alone expand
nonerythroid progenitor cells. A combination of 5 CSFs, ie, rIL-3, rIL-6,
rGM-CSF, rG-CSF, and rSCF, was identified as the most potent combination of
those tested and increased nonerythroid progenitor cells 57- +/- 11-fold.
After a 7-day suspension culture of CD34+ cells with these 5 CSFs, CD34+
cells expanded 14.5- fold, and CD34+/CD33- cells and CD34+/CD33+ cells were
also expanded 2.9-fold and 307-fold, respectively. Most secondary colonies
derived from expanded cells were small; however, the absolute number of
large- sized colonies expanded 5.9- +/- 3.3-fold. Thus, the combination of
CSFs can achieve a degree of amplification of PB CD34+ cells. The
capability of in vitro expansion of PB CD34+ cells as an adjunct to PB stem
cell transplantation is worthy of consideration.
Volume 82,
Issue 12,
pp. 3600-3609,
12/15/1993
Copyright © 1993 by The American Society of Hematology
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