Highly efficient mRNA-based gene transfer in feeder-free cultured H9 human embryonic stem cells |
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| Authors: | Ponsaerts Peter van der Sar Sjaak Van Tendeloo Viggo F I Jorens Philippe G Berneman Zwi N Singh Prim B |
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| Institution: | Laboratory of Experimental Hematology, Faculty of Medicine, Antwerp University Hospital, University of Antwerp, Antwerp, Belgium. Peter.Ponsaerts@ua.ac.be |
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| Abstract: | Several protocols have been described for virus-based gene transfer in human embryonic stem (hES) cells, while efficient non-viral methods are currently non-existing. In this study, we investigated the efficiency of mRNA-based gene transfer in feeder-free cultured H9 hES cells, based on electroporation of in vitro transcribed mRNA encoding the enhanced green fluorescent protein (EGFP). Optimisation of culture and electroporation conditions for feeder-free cultured H9 hES cells resulted a highly pure, transgene-expressing (90% positive cells) H9 hES cell population. |
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