Open AccessPromoter Dependence of Transgene Expression by Lentivirus‐Transduced Human Blood–Derived Endothelial Progenitor Cells
Author(s) -
Liu Jia Wei,
Pernod Gilles,
DunoyerGeindre Sylvie,
Fish Richard J.,
Yang Hong,
Bounameaux Henri,
Kruithof Egbert K. O.
Publication year - 2006
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2004-0364
Subject(s) - biology , transgene , progenitor cell , microbiology and biotechnology , genetic enhancement , peripheral blood mononuclear cell , viral vector , green fluorescent protein , promoter , transduction (biophysics) , lentivirus , transfection , stem cell , in vitro , gene , gene expression , immunology , recombinant dna , genetics , virus , biochemistry , viral disease
Peripheral blood– derived endothelial progenitor cells (EPCs) have considerable potential for the autologous therapy of vascular lesions or ischemic tissues. By introducing stable genetic modifications into these cells, this potential might be further enhanced. We investigated to what extent transgene expression can be controlled by using different transgene promoters. This was investigated in early‐ or late‐outgrowth human EPCs obtained by culturing blood mononuclear cells for 1 or 4 weeks on type 1 collagen in medium containing endothelial growth supplements. A large fraction of these cells were stably transduced using lentiviral vectors for expression of the enhanced green fluorescent protein (EGFP). Transgene expression in vitro or in vivo after injection into nude mice was highest when under the control of the cytomegalovirus (CMV) promoter, intermediate with the EF1α promoter, and lowest with the phosphoglycerate kinase promoter. When blood mononuclear cells were cultured for 1 week in the absence of endothelial growth supplements, CMV promoter– driven expression of EGFP was two orders of magnitude lower than in similarly transduced EPCs. Our results show that lentiviral vectors are useful tools for the stable introduction of exogenous genes into EPCs and for their expression at desired levels using the appropriate gene promoter.