Effect of Adenoviral Transduction of Hepatic Stellate Cells with Adv5-optHGF-RFP on their Phenotype

Abstract

Adenovirus is a promising gene delivery vector that has a high efficiency and relative ease of construction. These advantages make this system attractive for diverse research applications. In this project we performed transduction of hepatics stellate cells (HSC) with adenoviral vector containing hepatocytes growth factor (HGF) and red fluorescent protein (RFP) as a reporter (Adv5-optHGF-RFP), which let us to visualize the transduced cells. Further changes of phenotype were studied by real-time PCR and immunocytochemical staining. According to our results most of the cells were transduced and demonstrated stable expression of RFP. Effectiveness of the transduction was also confirmed by high expression of HGF (1200 times higher than in control HSC culture). Adenoviral transduction of HSC with HGF gene didn`t change their morphology, but stimulated expression of HSC marker desmin, lead to HSC activation (-SMA) and appearance of -FP, one of the hepatoblasts markers. During the entire ex