Polyplex-mediated gene transfer into human retinal pigment epithelial cells in vitro

Edward Chaum, Mark P. Hatton, Gary Stein

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The human retinal pigment epithelium (RPE) is a potential target tissue for directed transfer of candidate genes to treat age-related macular degeneration (AMD). The RPE is uniquely suited to gene therapy protocols that use liposome-mediated DNA transfer because of its high intrinsic phagocytic function in vivo. In these studies, we examined the efficacy of human RPE cell uptake and expression of the green fluorescent protein (GFP) and neomycin resistance marker genes by polyplex-mediated gene transfer in vitro. The effects of varying DNA and polyplex concentration and ratios on GFP transgene expression were examined. A narrow range of experimental conditions were found to maximize transgene expression; most important were the DNA concentration and the DNA:polyplex ratio. The transfection efficiency for human RPE cells was reproducibly 20% in vitro by this method and reached a maximum level of expression after 48 h. There was a rapid decline in gene expression over 2 weeks following polyplex-mediated gene transfer, but stable integration does occur at low frequencies with and without selection.

Original languageEnglish (US)
Pages (from-to)153-160
Number of pages8
JournalJournal of Cellular Biochemistry
Volume76
Issue number1
DOIs
StatePublished - Dec 1 1999

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Gene transfer
Retinal Pigments
Retinal Pigment Epithelium
Epithelial Cells
DNA
Green Fluorescent Proteins
Transgenes
Genes
Gene therapy
Neomycin
Macular Degeneration
Gene expression
Liposomes
Genetic Therapy
Transfection
Tissue
Gene Expression
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Polyplex-mediated gene transfer into human retinal pigment epithelial cells in vitro. / Chaum, Edward; Hatton, Mark P.; Stein, Gary.

In: Journal of Cellular Biochemistry, Vol. 76, No. 1, 01.12.1999, p. 153-160.

Research output: Contribution to journalArticle

Chaum, Edward ; Hatton, Mark P. ; Stein, Gary. / Polyplex-mediated gene transfer into human retinal pigment epithelial cells in vitro. In: Journal of Cellular Biochemistry. 1999 ; Vol. 76, No. 1. pp. 153-160.
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