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).
Similar content being viewed by others
References
Flotte TR, Laube BL. Gene therapy in cystic fibrosis. Chest. 2001;120(Suppl 3):124–131.
Dick CR, Ham GE. Characterisation of polyethylenimine. J Macromol Sci. 1970;4:1301–1314.
Klotz IM, Royer GP, Sloniewsky AR. Macromolecule-small molecule interactions: Strong binding and cooperativity in a model synthetic polymer. Biochemistry. 1969;8:4752–4756.
Boussif O, Lezoualch F, Zanta MA, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci U S A. 1995;92:7297–7301.
Godbey WT, Wu KK, Mikos AG. Poly(ethylenimine) and its role in gene delivery. J Control Release. 1999;60:149–160.
Godbey WT, Wu KK, Mikos AG. Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery. Proc Natl Acad Sci U S A. 1999;96:5177–5181.
Pollard H, Remy JS, Loussouarn G, Demolombe S, Behr JP, Escande D. Polyethylenimine but not cationic lipids promotes transgene delivery to the nucleus in mammalian cells. J Biol Chem. 1998;273:7507–7511.
Wiethoff CM, Smith JG, Koe GS, Middaugh CR. The potential role of proteoglycans in cationic lipid-mediated gene delivery: studies of the interaction of cationic lipid-DNA complexes with model glycosaminoglycans. J Biol Chem. 2001;276:32806–32813.
Lechardeur D, Sohn KJ, Haardt M, et al. Metabolic instability of plasmid DNA in the cytosol: a potential barrier to gene transfer. Gene Ther. 1999;6:482–497.
Moret I, Peris JE, Guillem VM, et al. Stability of PEI/DNA and DOTAP/DNA complexes: effect of alkaline pH, heparin, and serum. J Control Release. 2001;76:169–181.
Godbey WT, Wu KK, Mikos AG. Poly(ethylenimine)-mediated gene delivery affects endothelial cell function and viability. Biomaterials. 2001;22:471–480.
Putnam D, Gentry CA, Pack DW, Langer R. Polymer-based gene delivery with low cytotoxicity by a unique balance of side-chain termini. Proc Natl Acad Sci U S A. 2001;98:1200–1205.
Witschi C, Mrsny RJ. In vitro evaluation of microparticles and polymer gels for use as nasal platforms for protein delivery. Pharm Res. 1999;16:382–390.
Winton HL, Wan H, Cannell MB, et al. Cell lines of pulmonary and non-pulmonary origin as tools to study the effects of house dust mite proteinases on the regulation of epithelial permeability. Clin Exp Allergy. 1998;28:1273–1285.
Foster KA, Avery ML, Yazdanian M, Audus KL. Characterization of the Calu-3 cell line as a tool to screen pulmonary drug delivery. Int J Pharm. 2000;208:1–11.
Florea BI, van der Sandt I, Schrier SM, et al. Evidence of P-glycoprotein mediated apical to basolateral transport of flunisolide in human broncho-tracheal epithelial cells (Calu-3). Br J Pharmacol. 2001;134:1555–1563.
Meaney C, Florea BI, Borchard G, Junginger HE. Characterization of a human submucosal gland cell line (Calu-3) as in vitro model of the airway epithelium. Proc Int Symp Control Release Bioact Mater. 1999;26:198–199.
Ax W, Soldan M, Koch L, Maser E. Development of daunorubicin resistance in tumour cells by induction of carbonyl reduction. Biochem Pharmacol. 2000;59:293–300.
Holt PS, Buckley S, Deloach JR. Detection of the lethal effects of T-2 mycotoxin on cells using a rapid colorimetric viability assay. Toxicol Lett. 1987;39:301–312.
Lee TC, Wu R, Brody AR, Barrett JC, Nettesheim P. Growth and differentiation of hamster tracheal epithelial cells in culture. Exp Lung Res. 1984;6:27–45.
Guo W, Lee RJ. Efficient gene delivery via non-covalent complexes of folic acid and polyethylenimine. J Control Release. 2001;77:131–138.
Berger JT, Voynow JA, Peters KW, Rose MC. Respiratory carcinoma cell lines: MUC genes and glycoconjugates. Am J Respir Cell Mol Biol. 1999;20:500–510.
Lee MC, Penland CM, Widdicombe JH, Wine JJ. Evidence that Calu-3 human airway cells secrete bicarbonate. Am J Physiol. 1998;274:L450-L453.
Zhang Y, Doranz B, Yankaskas JR, Engelhardt JF. Genotypic analysis of respiratory mucous sulfation defects in cystic fibrosis. J Clin Invest. 1995;96:2997–3004.
Olavarrieta L, Martinez-Robles ML, Sogo JM, et al. Supercoiling, knotting and replication fork reversal in partially replicated plasmids. Nucleic Acids Res. 2002;30:656–666.
Von Harpe A, Petersen H, Li Y, Kissel T. Characterization of commercially available and synthesized polyethylenimines for gene delivery. J Control Release. 2000;69:309–322.
Matsuo A, Watanabe A, Takahashi T, et al. A simple method for classification of cell death by use of thin layer collagen gel for the detection of apoptosis and/or necrosis after cancer chemotherapy. Jpn J Cancer Res. 2001;92:813–819.
Kang YH, Chung SJ, Kang IJ, Park JH, Bunger R. Intramitochondrial pyruvate attenuates hydrogen peroxide-induced apoptosis in bovine pulmonary artery endothelium. Mol Cell Biochem. 2001;216:37–46.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published: July 9, 2002
Rights and permissions
About this article
Cite this article
Florea, B.I., Meaney, C., Junginger, H.E. et al. Transfection efficiency and toxicity of polyethylenimine in differentiated Calu-3 and nondifferentiated COS-1 cell cultures. AAPS J 4, 12 (2002). https://doi.org/10.1208/ps040312
Received:
Accepted:
Published:
DOI: https://doi.org/10.1208/ps040312