¹Corporate Development, Amaxa Biosystems, Cologne, Germany

^|[ast]|Presenter is employee of amaxa biosystems.

Abstract

Efficient non-viral transfection is a challenge, especially when it comes to difficult-to-transfect cell types like primary cells. For many years the performance of non-viral transfection technologies was lagging behind viral transduction. This situation has changed with the advent of the Nucleofector technology, an advanced electroporation method. Nucleofection is the first non-viral transfection technology that delivers DNA not only into the cytoplasm, but also straight into the nucleus of a cell. The Nucleofector technology routinely achieves transfection efficiencies of up to 90% with plasmid DNA and up to 99% with siRNA duplexes. The technology enables the highly efficient transfection of a broad variety of primary cell types including various kinds of stem cells. In numerous publications the potential of the technology has been shown with e.g. unstimulated T cells (transfection efficiency with DNA of up to 66 %) and stimulated T cells (47 %), chondrocytes (65 %), dendritic cells (49 %), neurons (59 %), fibroblasts (90 %), keratinocytes (53 %), CD34+ cells (71 %), mesenchymal stem cells (47 %), neural stem cells (60 %), and many others more.

Since its introduction the technology has opened up the doors to novel possibilities. While in the past most scientists made use of nucleofection in basic research and development, some scientists have already pursued the technology's potential regarding ex vivo gene therapeutic applications. Encouraged by their promising results, the technology will soon be available in a format that will allow for its use in clinical settings. Latest achievements using the Nucleofector technology in primary-cell transfection will be presented as well as latest information with respect to cGMP compliant versions of the Nucleofector technology.