Transfectability and Transducibility of chicken liver cell line LMH compared to human cell line HEK-293T

The aim of this study is to examine the efficiency of DNA absorption and gene transfusion to chicken hepatoma cells line LMH at both transfection and transduction levels using recombinant lentiviruses. Then transgene expression level was studied.

The cells were cultivated in 96-well plates for viral transfection or transduction and then studied using fluorescent microscope in different stages. Green Fluorescent Protein-positive (GFP) cells counted by Grid Cell Counter software and transgen expression level calculated using ImageJ software. Data analyzed using SPSS.

GFP gene expression began 6 and 9 hours after transfection in HEK and LMH cells, respectively. These times were increased, respectively, to 22 and 36 hours post-transduction. Counting GFP+ cells 48 hours post-transfection showed that HEK and LMH cells have received and expressed 40% and 35% of DNA. These levels increased to 95% and 48% respectively in 72 hours. Also counting HEK+ cells at 72 and 96 hours viral post-transduction confirmed the similar pattern of receiving DNA by both HEK and LMH cells. Therefore, analyzing of GFP expression level showed that HEK and LMH classes expressed GFP in 74% and 89% (P<0.01 significant).

Overall, our results indicated that HEK cells have higher potential to absorption of foreign genes whereas the intensity of expression is higher in LMH cells.