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

• Published in: Biotechnology letters Vol. 32; no. 10; pp. 1393 - 1399
• Main Authors: Madeira, C, Ribeiro, S. C, Turk, M. Z, Cabral, J. M. S
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01.10.2010; Springer Netherlands; Springer; Springer Nature B.V; Springer Verlag
• Subjects: Applied Microbiology; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Biotechnology - methods; Cell Line; Cell recovery; Cell Survival; Cells; Central Composite Design; Composite materials; Deoxyribonucleic acid; DNA; Electroporation - methods; Feasibility; Fundamental and applied biological sciences. Psychology; Gene delivery; Genes; Humans; Life Sciences; Methodology; Microbiology; Microporation; Optimization; Original Research Paper; Plasmid DNA; Recovery; Transfection; Viability; Central Composite Design; Gene delivery; Microporation; Plasmid DNA; Cell recovery; Design; Plasmid; DNA; Optimization; Genetic transfer
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-010-0327-4

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.