Transfection efficiency and toxicity of polyethylenimine in differentiated Calu-3 and nondifferentiated COS-1 cell cultures

Abstract

In the present study, we evaluated polyethylenimine (PEI) of different molecular weights (MWs) as a DNA complexing agent for its efficiency in transfecting nondifferentiated COS-1 (green monkey fibroblasts) and well-differentiated human submucosal airway epithelial cells (Calu-3). Studying the effect of particle size, zeta potential, presence of serum proteins or chloroquine, it appeared that transfection efficiency depends on the experimental conditions and not on the MW of the PEI used. Comparing transfection efficiencies in both cell lines, we found that PEI was 3 orders of magnitude more effective in COS-1 than in Calu-3 cells, because Calu-3 cells are differentiated and secrete mucins, which impose an additional barrier to gene delivery. Transfection efficiency was strongly correlated to PEI cytotoxicity. Also, some evidence for PEI-induced apoptosis in both cell lines was found. In conclusion, our results indicate that PEI is a useful vector for nonviral transfection in undifferentiated cell lines. However, results from studies in differentiated bronchial epithelial cells suggest that PEI has yet to be optimized for successful gene therapy of cystic fibrosis (CF).

Development of Dioctadecyldimethylammonium Bromide/Monoolein Liposomes for Gene Delivery

“…These molecules selfassemble in highly organized structures capable of complexing the genetic material and later releasing it inside the cells. Whereas viruses generally impose issues of mutagenicity and immunogenicity [16,17], polymers such as polyethylenimine (PEI) are known to be highly cytotoxic [18,19]. By exclusion of alternatives, cationic liposomes have emerged as morphology (lipoplex size, charge ratio (+/-), fluidity and structure) for optimal transfection conditions.…”

Ionic liquids-based processing of electrically conducting chitin nanocomposite scaffolds for stem cell growth

“…. However, the degraded products from polyurethane or polylactic acidbased scaffolds are known to cause inflammation [54][55][56] . The chitin/MWNT carbon nanotube composite can overcome this limitation due to the natural origin of chitin, its exceptionally low immunogenicity, antimicrobial activity and biocompatibility.…”

Microbeads mediated oral plasmid DNA delivery using polymethacrylate vectors: an effectual groundwork for colorectal cancer

“…Naked -gal pDNA showed very negligible transfection and was considered as negative control. PEI was taken as a standard non-viral polymer vector which has highest transfection ability at N/P 10 (Florea et al, 2002). DOTAP/DOPE was used as a standard non-viral lipid vectors which was taken at N/P 1, 2, 3, 4, 6, 8 and 10.…”

“…As -gal is not having any toxic effect, polyplex formed by this marker plasmid could be evaluated for toxicity of non-viral vector formulations. PEI was taken as a standard non-viral polymer vector which has highest transfection ability at N/P 10 (Florea et al, 2002) in HT29 cell line. Other vectors were prepared at different N/P ratios.…”