Mar 12, 2005

Neural retina limits the nonviral gene transfer to retinal pigment epithelium in an in vitro bovine eye model

The AAPS Journal
Leena PitkänenArto Urtti

Abstract

We investigated the permeation of liposomal and polymeric gene delivery systems through neural retina into retinal pigment epithelium (RPE) and determined the roles of various factors in permeation and subsequent uptake of the delivery systems by RPE. Anterior parts and vitreous of fresh bovine eyes were removed. Retina was left intact or peeled away. Complexes of ethidium monoazide (EMA)-labeled plasmid DNA and cationic carriers (polyethyleneimine, poly-L-lysine, DOTAP liposomes) were pipetted on the retina or RPE. Two hours later the neural retina was removed, if present, and the RPE cells were detached. Contaminants were removed by sucrose centrifugation, and the RPE cells were analyzed for DNA uptake by flow cytometry. Cellular uptake of FITC-dextrans (molecular weight [mw] 20,000, 500,000 and 2,000,000), FITC-poly-L-lysine (mw 20,000), FITC-labeled oligonucleotide (15-mer), and naked EMA-labeled plasmid DNA was determined after pipetting the solutions on the RPE or neural retina. Location of the fluorescent materials in the retina was visualized with fluorescence microscopy. Neural retina decreased the cellular uptake of DNA complexes by an order of magnitude, the uptake of FITC-dextrans slightly, whereas delivery of polyc...Continue Reading

  • References26
  • Citations21

References

  • References26
  • Citations21

Citations

Mentioned in this Paper

Flow Cytometry
Gene Transfer Techniques
1,2-dioleoyloxy-3-(trimethylammonium)propane
Entire Retina
Pigment Epithelium of Eye
Complex (molecular entity)
Fluorescence-Activated Cell Sorting
Uptake
Bos taurus
Retinal Diseases

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