Optimization of gene delivery to HEK293T cells by microporation using a central composite design methodology.
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.
Madeira
C
C
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
Ribeiro
S C
SC
Turk
M Z
MZ
Cabral
J M S
JM
D001709:Biotechnology / Q000379:methods*; D002460:Cell Line; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D006801:Humans; D014162:Transfection