Live fluorescent RNA-based detection of pluripotency gene expression in embryonic and induced pluripotent stem cells of different species

Harald Lahm, Stefanie Doppler, Martina Dreßen, Astrid Werner, Klaudia Adamczyk, Dominic Schrambke, Thomas Brade, Karl Ludwig Laugwitz, Marcus André Deutsch, Matthias Schiemann, Rüdiger Lange, Alessandra Moretti, Markus Krane

Research output: Contribution to journalArticlepeer-review

26 Scopus citations


The generation of induced pluripotent stem (iPS) cells has successfully been achieved in many species. However, the identification of truly reprogrammed iPS cells still remains laborious and the detection of pluripotency markers requires fixation of cells in most cases. Here, we report an approach with nanoparticles carrying Cy3-labeled sense oligonucleotide reporter strands coupled to gold-particles. These molecules are directly added to cultured cells without any manipulation and gene expression is evaluated microscopically after overnight incubation. To simultaneously detect gene expression in different species, probe sequences were chosen according to interspecies homology. With a common target-specific probe we could successfully demonstrate expression of the GAPDH house-keeping gene in somatic cells and expression of the pluripotency markers NANOG and GDF3 in embryonic stem cells and iPS cells of murine, human, and porcine origin. The population of target gene positive cells could be purified by fluorescence-activated cell sorting. After lentiviral transduction of murine tail-tip fibroblasts Nanog-specific probes identified truly reprogrammed murine iPS cells in situ during development based on their Cy3-fluorescence. The intensity of Nanog-specific fluorescence correlated positively with an increased capacity of individual clones to differentiate into cells of all three germ layers. Our approach offers a universal tool to detect intracellular gene expression directly in live cells of any desired origin without the need for manipulation, thus allowing conservation of the genetic background of the target cell.

Original languageEnglish
Pages (from-to)392-402
Number of pages11
JournalStem Cells
Issue number2
StatePublished - 1 Feb 2015


  • Embryonic stem cells
  • Fluorescent nanoparticles
  • Induced pluripotent stem cells
  • Live staining
  • Pluripotency


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