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

• Main Authors: Alimohammad Asgharian, Mehdi Banan, Hossein Najmabadi
• Format: Article
• Language: English
• Published: Royan Institute (ACECR), Tehran 2013-01-01
• Series: Cell Journal
• Subjects: Transfection; HeLa Cells; Lipids
• Online Access: http://celljournal.org/library/upload/article/af_572534623272666234776225455236564524243613-Asgharian-1.pdf

One of the most significant steps in gene expression studies is transferring genes into cell cultures. Despite there are different methods for gene delivery such as viral and non-viral producers, some cationic lipid reagents have recently developed to transfect into mammalian cell lines. The main aim of this study was optimizing and improving lipocomplex based transient transfection procedures into HeLa cell line which is being used widely as a typical cell in biological studies.This study was an experimental research. In this work, pCMV: β-Gal DNA plasmid was used as a reporter DNA for determining the rate of gene transfection into HeLa cells. To accomplish the highest gene delivery into HeLa cells, optimizing experiments were carried out in different volumes of FuGENE-HD, LipofectamineTM2000 and X-tremeGENE. Also, we investigated tranasfection efficiency in presence of various cell densities of HeLa cells. Then, transfection efficiency and cell toxicity were measured by beta gal staining and trypan blue methods, respectively.Using FuGENE-HD in volume of 4μl along with 105 HeLa cells, transfection efficiency was higher (43.66 ± 1.52%) in comparison with the cationic lipids lipofectamineTM2000 and X-tremeGENE. In addition, the rate of cell toxicity in presence of FuGENE-HD was less than 5%.In summary, the cationic lipid FuGENE-HD indicates a suitable potential to transfer DNA into 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 cell lines with FuGENE-HD due to its low toxicity and high efficiency.