Recombinant lentivirus-mediated gene transfer into chicken cell line LMH

Lentiviruses are considered one of the most effective recombinant viruses for gene transfer to mammalian cells and tissues. In this study, the potential of HIV-1-based lentiviral vector to deliver transgenes into avian cells was examined. We co-transfected human embryonic kidney cell line HEK-293T with three lentivirus vectors called transfer, packaging and envelope vectors. We collected the supernatant from transfected cells 24 and 48 hours post-transfection and filtered them immediately. Then we subjected the filtered supernatant to Amicon protein columns for concentration purposes. Centrifugation removed a larger part of the supernatant presumably free of viruses and left behind a small volume of darken solution full of virions. We thereby produced a 500-µl-volume of virus stock. Various dilutions of this stock were added to chicken liver cell line LMH. The initial sign of infection appeared within 48 hours and by 96 hours post-infection 100% the LMH cells positively expressed transgenes. Our results indicated that the human HIV-1-based lentivirus vectors are capable of transducing and transferring foreign genes into chicken cells. Given the need for a high-titer virus stock for successful target cell transduction, our results indicate that the filtration method of virus concentration is able to produce high virus titer and is cost-effective and less time consuming than ultracentrifugation or other traditional methods.