Optimization of gene delivery to HEK293T cells by microporation using a central composite design methodology

C. Madeira (1) , S. C. Ribeiro (1) , M. Z. Turk (1) , J. M. S. Cabral (1)

1 IBB - Institute for Biotechnology and Bioengineering

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.

Mots clés: Cell recovery; Central Composite Design; Gene delivery; Microporation; Plasmid DNA

C. Madeira, S. C. Ribeiro, M. Z. Turk, J. M. S. Cabral. Optimization of gene delivery to HEK293T cells by microporation using a central composite design methodology. Biotechnology Letters, 2010, 32 (10), pp.1393-1399. ⟨10.1007/s10529-010-0327-4⟩. ⟨hal-00601524⟩