Highly efficient mRNA-based gene transfer in feeder-free cultured H9 human embryonic stem cells

Cloning Stem Cells. 2004;6(3):211-6. doi: 10.1089/clo.2004.6.211.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Survival / physiology*
  • Cells, Cultured
  • Electroporation / methods*
  • Flow Cytometry
  • Green Fluorescent Proteins / genetics*
  • Humans
  • Stem Cells / cytology*
  • Transfection / methods

Substances

  • Green Fluorescent Proteins