Transfection efficiency and toxicity of polyethylenimine in differentiated Calu-3 and nondifferentiated COS-1 cell cultures.
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).
Florea
Bogdan I
BI
Division of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, The Netherlands.
Meaney
Clare
C
Junginger
Hans E
HE
Borchard
Gerrit
G
D000818:Animals; D006690:Bisbenzimidazole / Q000494:pharmacology; D019556:COS Cells / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004247:DNA / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D007820:Laryngeal Mucosa / Q000166:cytology / Q000187:drug effects*; D009690:Nucleic Acid Conformation; D011094:Polyethyleneimine / Q000378:metabolism / Q000633:toxicity*; D013194:Staining and Labeling / Q000379:methods; D014162:Transfection / Q000379:methods* / Q000592:standards*
D005456:Fluorescent Dyes; D011094:Polyethyleneimine; D004247:DNA; D006690:Bisbenzimidazole