<  Back to the Polytechnique Montréal portal

Enhanced gene delivery mediated by low molecular weight chitosan/DNA complexes: effect of pH and serum

Surendra Nimesh, Marc Thibault, Marc Lavertu and Michael D. Buschmann

Article (2010)

[img]
Preview
Published Version
Terms of Use: Creative Commons Attribution Non-commercial .
Download (904kB)
Cite this document: Nimesh, S., Thibault, M., Lavertu, M. & Buschmann, M. D. (2010). Enhanced gene delivery mediated by low molecular weight chitosan/DNA complexes: effect of pH and serum. Molecular Biotechnology, 46(2), p. 182-96. doi:10.1007/s12033-010-9286-1
Show abstract Hide abstract

Abstract

This study was designed to systematically evaluate the influence of pH and serum on the transfection process of chitosan-DNA complexes, with the objective of maximizing their efficiency. The hydrodynamic diameter of the complexes, measured by dynamic light scattering (DLS), was found to increase with salt and pH from 243 nm in water to 1244 nm in PBS at pH 7.4 and aggregation in presence of 10% serum. The cellular uptake of complexes into HEK 293 cells assessed by flow cytometry and confocal fluorescent imaging was found to increase at lower pH and serum. Based on these data, new methodology were tested and high levels of transfection (>40%) were achieved when transfection was initiated at pH 6.5 with 10% serum for 8-24 h to maximize uptake and then the media was changed to pH 7.4 with 10% serum for an additional 24-40 h period. Cytotoxicity of chitosan/DNA complexes was also considerably lower than Lipofectamine. Our study demonstrates that the evaluation of the influence of important parameters in the methodology of transfection enables the understanding of crucial physicochemical and biological mechanisms which allows for the design of methodologies maximising transgene expression.

Uncontrolled Keywords

Animals; Cattle; *Chitosan; DNA/*administration & dosage/*metabolism; *Gene Transfer Techniques; HEK293 Cells; Humans; Hydrogen-Ion Concentration; Luciferases/analysis; Transfection/*methods

Open Access document in PolyPublie
Department: Département de génie chimique
Date Deposited: 05 Apr 2022 14:46
Last Modified: 06 Apr 2022 01:20
PolyPublie URL: https://publications.polymtl.ca/5060/
Document issued by the official publisher
Journal Title: Molecular Biotechnology (vol. 46, no. 2)
Publisher: Springer
Official URL: https://doi.org/10.1007/s12033-010-9286-1

Statistics

Total downloads

Downloads per month in the last year

Origin of downloads

Dimensions

Repository Staff Only