Lentiviral transfer of the LacZ gene into human endothelial cells and human bone marrow mesenchymal stem cells.
Because one of the attractive characteristics of human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors is that it can infect even nondividing cells, a lentivirus-mediated gene delivery system is currently being paid a great deal of attention as an innovative tool for gene transfer into target cells. The purpose of the work was to investigate the efficacy of lentiviral transfer of the LacZ gene into human umbilical vein endothelial cells (HUVECs) and human bone marrow mesenchymal stem cells (HMSCs) in vitro. For the present study, a vesicular stomatitis virus G-protein (VSV-G)-pseudotyped lentiviral vector encoding the E. coli LacZ gene tagged with nuclear localization signal (NLS) was generated in 293T cells by means of the three-plasmid system. The resulting lentiviral vector, LtV-NLS/LacZ, was allowed to infect HUVECs and HMSCs. Approximately 70% of HUVECs were positive for LacZ expression and 50% of HMSCs showed LacZ activity. There was no significant difference in transduction efficacy between early and late-passage phases in both cells. LtV-NLS/LacZ-transduced HUVECs showed gene expression of endothelial markers including CD34 and flt-1 and KDR/flk-1 of vascular endothelial growth factor (VEGF) receptors and had angiogenic potential as efficiently as primarily cultured HUVECs in a Matrigel assay. These findings provide evidence that lentiviral vectors are efficient tools for gene transfer and expression in human endothelial cells and stem cells that could be useful for tissue engineering.
Totsugawa
Toshinori
T
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Japan.
Kobayashi
Naoya
N
Okitsu
Teru
T
Noguchi
Hirofumi
H
Watanabe
Takamasa
T
Matsumura
Toshihisa
T
Maruyama
Masanobu
M
Fujiwara
Toshiyoshi
T
Sakaguchi
Masakiyo
M
Tanaka
Noriaki
N
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D003094:Collagen; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000821:virology; D005786:Gene Expression Regulation; D006801:Humans; D007763:Lac Operon; D007797:Laminin; D016086:Lentivirus / Q000235:genetics*; D008648:Mesoderm / Q000166:cytology*; D018919:Neovascularization, Physiologic; D010710:Phosphates / Q000737:chemistry; D011509:Proteoglycans; D051381:Rats; D013234:Stem Cells / Q000378:metabolism* / Q000821:virology; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics* / Q000378:metabolism
D004338:Drug Combinations; D007797:Laminin; D010710:Phosphates; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D001616:beta-Galactosidase