Gene delivery through cell culture substrate adsorbed DNA complexes

Zain Bengali, Angela K Pannier, Tatiana Segura, Brian C. Anderson, Jae Hyung Jang, Thomas A. Mustoe, Lonnie D. Shea

Research output: Contribution to journalArticle

106 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)290-302
Number of pages13
JournalBiotechnology and Bioengineering
Volume90
Issue number3
DOIs
StatePublished - May 5 2005
Externally publishedYes

Fingerprint

Cell culture
Polystyrenes
Biocompatible Materials
DNA
Cell Culture Techniques
Genes
Substrates
Tissue culture
Plasmids
Serum
Immobilized Nucleic Acids
Biomaterials
Cellular Microenvironment
Polyglactin 910
Immobilized Cells
Tissue Engineering
Biotechnology
Reporter Genes
Immobilization
Adsorption

Keywords

  • Biomaterial
  • Gene delivery
  • Lipofectamine 2000
  • Polyethylenimine
  • Reverse transfection
  • Solid-phase delivery
  • Substrate-mediated

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Bengali, Z., Pannier, A. K., Segura, T., Anderson, B. C., Jang, J. H., Mustoe, T. A., & Shea, L. D. (2005). Gene delivery through cell culture substrate adsorbed DNA complexes. Biotechnology and Bioengineering, 90(3), 290-302. https://doi.org/10.1002/bit.20393

Gene delivery through cell culture substrate adsorbed DNA complexes. / Bengali, Zain; Pannier, Angela K; Segura, Tatiana; Anderson, Brian C.; Jang, Jae Hyung; Mustoe, Thomas A.; Shea, Lonnie D.

In: Biotechnology and Bioengineering, Vol. 90, No. 3, 05.05.2005, p. 290-302.

Research output: Contribution to journalArticle

Bengali, Z, Pannier, AK, Segura, T, Anderson, BC, Jang, JH, Mustoe, TA & Shea, LD 2005, 'Gene delivery through cell culture substrate adsorbed DNA complexes', Biotechnology and Bioengineering, vol. 90, no. 3, pp. 290-302. https://doi.org/10.1002/bit.20393
Bengali, Zain ; Pannier, Angela K ; Segura, Tatiana ; Anderson, Brian C. ; Jang, Jae Hyung ; Mustoe, Thomas A. ; Shea, Lonnie D. / Gene delivery through cell culture substrate adsorbed DNA complexes. In: Biotechnology and Bioengineering. 2005 ; Vol. 90, No. 3. pp. 290-302.
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