Nucleofection has been successfully used for generation of iPS cells

Capsugel Australia Pty Ltd
Tuesday, 07 September, 2010

Reprogramming of somatic cells to pluripotent iPS (induced pluripotent stem) cells promises to revolutionize regenerative medicine.

Up to now mainly viral vectors have been used for reprogramming experiments. Now safety issues related to viral transduction methods can be circumvented by using Nucleofection:

Kaji K, Norrby K, Paca A, Mileikovsky M, Mohseni P, Woltjen K. Virus-free induction of pluripotency and subsequent excision of reprogramming factors. Nature 2009 Apr 9;458(7239):771-5. Epub 2009 Mar 1.

Nucleofection of a single expression vector comprising the coding sequences of 4 transcription factors (c-Myc, Klf4, Oct4 and Sox2) into mouse embryonic fibroblasts (MEFs)
Reprogramming of fibroblasts confirmed by robust expression of pluripotency markers, in vitro differentiation assays and formation of adult chimeric mice
After generation of iPS cells, excision of expression cassette (Cre-mediated)

See additional data (not published) to the left.

MEF cells were stably transfected with a vector encoding for transcription factors c-Myc, Klf4, Oct4 and Sox2 and EGFP. 2 days post Nucleofection, cells were stained for Oct4 and Sox2 and analyzed for GFP expression.

8 days post Nucleofection, cells show ES cell like morphology and were analyzed for GFP expression. All data kindly provided by and published with permission of Keisuke Kaji, Institute for Stem Cell research, University of Edinburgh, UK.