Retroviral vector-mediated transfer of the bacterial neomycin resistance
gene into fetal and adult sheep and human hematopoietic progenitors in
vitro
D Ekhterae, T Crumbleholme, E Karson, MR Harrison, WF Anderson and ED Zanjani
Department of Veterans' Affairs Medical Center, University of Nevada School
of Medicine, Reno.
We compared the efficiency of retroviral vector (N2)-mediated transfer of
the bacterial neomycin resistance gene (NeoR) into adult and fetal
hematopoietic progenitors of sheep and humans by assessing their ability to
form colonies in the presence of lethal doses of the neomycin analogue G418
in vitro. Fetal cells from both sheep and humans exhibited a higher degree
of NeoR transfer than adult cells. The overall level of NeoR expression was
significantly higher for sheep than human cells. The transfer/expression of
NeoR into adult human bone marrow hematopoietic progenitors was not
affected by the presence or absence of T cells and monocyte/macrophages.
The efficiency of NeoR transfer into both adult and fetal human cells,
however, was improved when transduction was carried out in the presence of
recombinant human interleukin-3 and granulocyte-macrophage
colony-stimulating factor. These results demonstrate the greater efficiency
of NeoR gene transfer into fetal hematopoietic progenitors, which may
provide a basis for the relatively higher efficiency of the in utero
approach to gene therapy.
Volume 75,
Issue 2,
pp. 365-369,
01/15/1990
Copyright © 1990 by The American Society of Hematology
