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Retroviral transfer of the recombinant human erythropoietin receptor gene
into single hematopoietic stem/progenitor cells from human cord blood
increases the number of erythropoietin-dependent erythroid colonies
L Lu, Y Ge, ZH Li, W Keeble, D Kabat, GC Bagby, HE Broxmeyer and ME Hoatlin
Department of Medicine (Hematology/Oncology), Indiana University School of
Medicine, Indianapolis 46202-5121, USA.
To test whether an enforced expression of a lineage-specific cytokine
receptor would influence the proliferation/differentiation of hematopoietic
stem/progenitor cells, retroviral vectors containing the human
erythropoietin receptor (hEpoR) gene were used to transduce the hEpoR gene
into phenotypically sorted subsets of cells. CD34 , CD34++CD33-, and
CD34++CD33+ populations of human cord blood, highly enriched for
hematopoietic stem/progenitor cells, were sorted and plated as single cells
per well in methylcellulose culture medium containing early acting growth
factors in the presence or absence of Epo. The hEpoR gene was efficiently
transduced into single high proliferative potential colony-forming cells
(HPP-CFC) and multipotential (colony-forming unit granulocyte, erythroid,
monocyte, megakaryocyte [CFU-GEMM]), erythroid (burst-forming
unit-erythroid [BFU- E]), and granulocyte-macrophage (colony-forming
unit-granulocyte- macrophage [CFU-GM]) progenitor cells. As expected in
cultures grown in the absence of Epo, no BFU-E or CFU-GEMM colonies grew.
In the presence of Epo, the hEpoR-gene transduced cells formed
significantly more CFU- GEMM and BFU-E colonies than did the controls. A
significant decrease in HPP-CFC colonies was also observed under these
conditions. Little or no effect of hEpoR gene transduction was apparent in
the numbers of CFU- GM colonies formed in the presence or absence of Epo.
All of the above results were similar whether the cell populations assessed
were CD34 or their CD33- or CD33+ subsets plated in the presence of growth
factors at 200 cells/mL or after limiting dilution at 2 cells/well. These
results suggest that the profile of detectable stem/progenitors can be
altered by retrovirus-mediated expression of the hEpoR gene.
Volume 87,
Issue 2,
pp. 525-534,
01/15/1996
Copyright © 1996 by The American Society of Hematology
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