Lentiviral vector: a useful tool for transduction of human liver endothelial cells.
Endothelial cells play multiple roles in pathophysiologic processes and are increasingly being recognized as target cells of gene therapy. Lentiviral vectors derived from human immunodeficiency virus type 1 have an ability to infect both dividing and nondividing cells and currently receive a great deal of attention as an innovative tool for transduction of target cells. The purpose of the present work was to evaluate the efficacy of a lentiviral vector for transducing human liver endothelial cells (HLECs) in vitro. For the present study, a pseudotyped lentiviral vector encoding a green fluorescent protein (GFP) gene, LtV-GFP, was generated by means of FuGENE 6 method and allowed to infect HLECs. Approximately 95% of HLECs were positive for GFP expression after LtV-GFP infection at a multiplicity of infection of 10. Notably, LtV-GFP transduced HLECs had stable and long term GFP expression, showed gene expression of endothelial markers including CD 34, factor VIII, flt-1, KDR/flk-1 and HGF, and maintained in vitro angiogenic potential in a Matrigel assay to the same extent as primarily cultured HLECs. These findings provide evidence that lentivirus based gene delivery is an efficient tool for transduction of endothelial cells that could be considered for cell and gene therapies and hybrid artificial organs.
Totsugawa
Toshinori
T
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Japan.
Kobayashi
Naoya
N
Maruyama
Masanobu
M
Kosaka
Yoshikazu
Y
Okitsu
Teru
T
Arata
Takashi
T
Sakaguchi
Masakiyo
M
Ueda
Tadayoshi
T
Kurabayashi
Yuzuru
Y
Tanaka
Noriaki
N
D001672:Biocompatible Materials; D003094:Collagen; D004338:Drug Combinations; D042783:Endothelial Cells / Q000502:physiology*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D007797:Laminin; D008099:Liver / Q000166:cytology*; D008164:Luminescent Proteins / Q000235:genetics; D018919:Neovascularization, Physiologic; D011509:Proteoglycans; D017186:Titrimetry; D014161:Transduction, Genetic / Q000379:methods*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*
D001672:Biocompatible Materials; D004338:Drug Combinations; D007797:Laminin; D008164:Luminescent Proteins; D011509:Proteoglycans; C059485:matrigel; D049452:Green Fluorescent Proteins; D003094:Collagen