Optimizing A Lipocomplex-Based Gene Transfer Method into HeLa Cell Line

One of the most significant steps in gene expression studies is transferring genes into cellcultures. despite there are different methods for gene delivery such as viral and non-viralproducers, some cationic lipid reagents have recently developed to transfect into mammaliancell lines. the main aim of this study was optimizing and improving lipocomplexbased transient transfection procedures into hela cell line which is being used widely asa typical cell in biological studies. this study was an experimental research. in this work, pcmv: b-gal dna plasmid wasused as a reporter dna for determining the rate of gene transfection into hela cells. toaccomplish the highest gene delivery into hela cells, optimizing experiments were carriedout in different volumes of fugene-hd, lipofectamine2000 and x-tremegene.also, we investigated tranasfection efficiency in presence of various cell densities of helacells. then, transfection efficiency and cell toxicity were measured by beta gal stainingand trypan blue methods, respectively. using fugene-hd in volume of 4µl along with 105tm hela cells, transfection efficiency was higher (43.66 ± 1.52%) in comparison with the cationic lipids lipofectamine2000and x-tremegene. in addition, the rate of cell toxicity in presence of fugene-hd wasless than 5%.in summary, the cationic lipid fugene-hd indicates a suitable potential to transfer dnainto hela cells and it can be an efficient reagent for gene delivery for hela cells in vitro.moreover, it is worth designing and optimizing gene transfer experiments for other celllines with fugene-hd due to its low toxicity and high efficiency. tm