Programm                 "Degeneration und Regeneration– Grundlagen, Diagnostik und Therapie"


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Abstract
Abstract

Efficient Non-viral Transfection of Neural Stem Cells

Richard I.1, Ader M.1, Sytnyk V.2, Richard G.1, Schachner M.2, Bartsch U.2 
1University Eye Clinic Hamburg Eppendorf and 2Zentrum für Molekulare Neurobiologie, Hamburg

Purpose: Cell therapy (i.e. replacement of degenerated or dysfunctional cells) with neural stem cells (NSCs) is discussed as a possible strategy to treat neurodegenerative disorders. In addition, transplantations of genetically engineered NSCs might be of use to target therapeutic gene products to the diseased nervous system. Here, we describe a novel non-viral transfection technique which allows the efficient transfection of in vitro expanded NSCs derived from the spinal cord of mouse embryos.
Method: NSCs were isolated from the spinal cord of wild-type, L1-deficient or enhanced green fluorescent protein- (EGFP) transgenic mouse embryos, and expanded in vitro in the presence of epidermal growth factor and fibroblast growth factor-2. Cells from the third to sixth passage were transfected with the reporter gene EGFP or the cell recognition molecule L1 using the NucleofectorTM technique (Amaxa Biosystems, Köln, Germany). Transfected cells were analysed between one day and 3.5 weeks after transfection in vitro, and up to 3.5 weeks after intraretinal transplantation in vivo.
Results: Quantitative analysis of wild-type cultures and L1-deficient cultures one day after transfection with EGFP and L1, respective


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