Journal of Stem Cell Research & Therapy

Safe and efficient gene transfer to retinal cells via a novel DOTMA-based niosomal vector

4^th International Conference and Exhibition on Cell & Gene Therapy

August 10-12, 2015 London, UK

Noha Attia

Scientific Tracks Abstracts: J Stem Cell Res Ther

Abstract:

Niosomes represent a recent promising approach for gene delivery purposes. Our group formulated a novel cationic niosome
based on N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA) cationic lipid combined with
polysorbate 60 and lycopene to evaluate the transfection efficiency in retinal pigment cells. Cationic niosomes prepared by
emulsification-evaporation technique were mixed with the anionic pCMS-EGFP plasmid to form by electrostatic interaction
nioplexes which were characterized in terms of morphology, size, surface charge and ability for DNA condensation, release and
protection. In vitro studies were conducted to evaluate cell uptake, cytoplasmic distribution, transfection efficiency and viability
in ARPE-19 cells. Nioplexes at 18/1 w/w ratio (cationic lipid/plasmid ratio) were 94 nm in size, +27 mV in zeta potential and
exhibited imperfectly spherical morphology. At such mass ratio, niosomes condensed, released and protected the DNA from
enzymatic digestion. When compared with commercial Lipofectamine 2000 lipoplexes, our nioplexes efficiently transfected
ARPE-19 cells without hampering their viability. Nioplexes were quickly internalized and homogeneously distributed in
the cytoplasm of cells. Furthermore, our results show that macropinocytosis, caveola-mediated endocytosis and to lesser
extent clathrin-dependent endocytosis, all contribute to nioplexes uptake and transport into lysosomes. This novel niosome
formulation represents a promising approach to deliver genetic material into the retina to treat inherited retinal diseases.

Biography :

Noha Attia has completed her MD in 2014 from Alexandria University, Egypt and Postdoctoral studies at University of the Basque Country, Spain. She is a Lecturer at Histology
& Cell Biology Department, Alexandria Medical School and the Manager of Tissue Culture Lab of Alexandria Center of Excellence in Regenerative Medicine. She is the PI
of research project entitled “Gene transfection of mesenchymal stem cells in 3D culture using non-viral vectors” funded by Science & Technology Development Fund, 2015.