Highly efficient mRNA-based gene transfer in feeder-free cultured H9 human embryonic stem cells.
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.
Ponsaerts
Peter
P
Laboratory of Experimental Hematology, Faculty of Medicine, Antwerp University Hospital, University of Antwerp, Antwerp, Belgium. Peter.Ponsaerts@ua.ac.be
van der Sar
Sjaak
S
Van Tendeloo
Viggo F I
VF
Jorens
Philippe G
PG
Berneman
Zwi N
ZN
Singh
Prim B
PB
D002470:Cell Survival / Q000502:physiology*; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D013234:Stem Cells / Q000166:cytology*; D014162:Transfection / Q000379:methods
D049452:Green Fluorescent Proteins