Article

Gene delivery through cell culture substrate adsorbed DNA complexes.

Department of Interdepartmental Biological Sciences, Northwestern University, 2145 Sheridan Rd./E156, Evanston, Illinois 60208-3120, USA.
Biotechnology and Bioengineering (impact factor: 3.95). 06/2005; 90(3):290-302. DOI:10.1002/bit.20393 pp.290-302
Source: PubMed

ABSTRACT Efficient gene delivery is a fundamental goal of biotechnology and has numerous applications in both basic and applied science. Substrate-mediated delivery and reverse transfection enhance gene transfer by increasing the concentration of DNA in the cellular microenvironment through immobilizing a plasmid to a cell culture substrate prior to cell seeding. In this report, we examine gene delivery of plasmids that were complexed with cationic polymers (polyplexes) or lipids (lipoplexes) and subsequently immobilized to cell culture or biomaterial substrates by adsorption. Polyplexes and lipoplexes were adsorbed to either tissue culture polystyrene or serum-adsorbed tissue culture polystyrene. The quantity of DNA immobilized increased with time of exposure, and the deposition rate and final amount deposited depended upon the properties of the substrate and complex. For polyplexes, serum modification enhanced reporter gene expression up to 1500-fold relative to unmodified substrates and yielded equivalent or greater expression compared to bolus delivery. For lipoplexes, serum modification significantly increased the number of transfected cells relative to unmodified substrates yet provided similar levels of expression. Immobilized complexes transfect primary cells with improved cellular viability relative to bolus delivery. Finally, this substrate-mediated delivery approach was extended to a widely used biomaterial, poly(lactide-co-glycolide). Immobilization of DNA complexes to tissue culture polystyrene substrates can be a useful tool for enhancing gene delivery for in vitro studies. Additionally, adapting this system to biomaterials may facilitate application to fields such as tissue engineering.

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Keywords

biomaterial substrates
 
bolus delivery
 
cell culture substrate
 
cell seeding
 
deposition rate
 
DNA immobilized
 
Efficient gene delivery
 
final amount
 
gene delivery
 
gene transfer
 
reverse transfection
 
serum-adsorbed tissue culture polystyrene
 
Substrate-mediated delivery
 
substrate-mediated delivery approach
 
tissue culture polystyrene substrates
 
tissue engineering
 
transfected cells
 
unmodified substrates
 
used biomaterial
 
useful tool