title: Optimization of gene delivery to HEK293T cells by microporation using a central composite design methodology.
abstract: Microporation is an efficient method for delivering plasmid DNA molecules into cultured cells. Herein, we present the optimization of gene delivery by microporation using a Central Composite Design methodology. It was given relevance not only to the transfection efficiency but also to the cell recovery. Different amounts of DNA (1 and 3 μg) mainly affected cell viabilities and cell recoveries, which decrease from 93 to 76% and from 47 to 25% respectively, when higher DNA quantity is used. With this work we suggest an easy methodology to improve transfection of mammalian cells underlining the feasibility to achieve 60% of gene delivery efficiencies whilst recovering 50% of cells, with 90% of viability.
authors.affiliation: IBB-Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001, Lisbon, Portugal. catarina.madeira@ist.utl.pt
authors.forename: C
authors.initials: C
authors.lastname: Madeira
authors.forename: S C
authors.initials: SC
authors.lastname: Ribeiro
authors.forename: M Z
authors.initials: MZ
authors.lastname: Turk
authors.forename: J M S
authors.initials: JM
authors.lastname: Cabral
mesh_terms: D001709:Biotechnology / Q000379:methods*; D002460:Cell Line; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D006801:Humans; D014162:Transfection