title: The Comparative Utility of Viromer RED and Lipofectamine for Transient Gene Introduction into Glial Cells.
abstract: The introduction of genes into glial cells for mechanistic studies of cell function and as a therapeutic for gene delivery is an expanding field. Though viral vector based systems do exhibit good delivery efficiency and long-term production of the transgene, the need for transient gene expression, broad and rapid gene setup methodologies, and safety concerns regarding in vivo application still incentivize research into the use of nonviral gene delivery methods. In the current study, aviral gene delivery vectors based upon cationic lipid (Lipofectamine 3000) lipoplex or polyethylenimine (Viromer RED) polyplex technologies were examined in cell lines and primary glial cells for their transfection efficiencies, gene expression levels, and toxicity. The transfection efficiencies of polyplex and lipoplex agents were found to be comparable in a limited, yet similar, transfection setting, with or without serum across a number of cell types. However, differential effects on cell-specific transgene expression and reduced viability with cargo loaded polyplex were observed. Overall, our data suggests that polyplex technology could perform comparably to the market dominant lipoplex technology in transfecting various cells lines including glial cells but also stress a need for further refinement of polyplex reagents to minimize their effects on cell viability.
authors.affiliation: The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
authors.forename: Sudheendra
authors.initials: S
authors.lastname: Rao
authors.affiliation: The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
authors.forename: Alejo A
authors.initials: AA
authors.lastname: Morales
authors.affiliation: The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA ; The Departments of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL 33136, USA ; The Neuroscience Program, University of Miami Miller School of Medicine, Miami, FL 33136, USA ; The Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
authors.forename: Damien D
authors.initials: DD
authors.lastname: Pearse
mesh_terms: D000818:Animals; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry* / Q000235:genetics; D009457:Neuroglia / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D014162:Transfection / Q000379:methods
chemical_list: D008055:Lipids; C086724:Lipofectamine