Derivation and characterization of sleeping beauty transposon-mediated porcine induced pluripotent stem cells

Research output: Contribution to journalJournal articleResearchpeer-review

  • Wilfried A. Kues
  • Doris Herrmann
  • Brigitte Barg-Kues
  • Srividiameena Haridoss
  • Monika Nowak-Imialek
  • Thomas Buchholz
  • Miriam Streeck
  • Antonia Grebe
  • Ivana Grabundzija
  • Sylvia Merkert
  • Ulrich Martin
  • Hall, Vanessa Jane
  • Mikkel Aabech Rasmussen
  • Zoltan Ivics
  • Poul Hyttel
  • Heiner Niemann
The domestic pig is an important large animal model for preclinical testing of novel cell therapies. Recently, we produced pluripotency reporter pigs in which the Oct4 promoter drives expression of the enhanced green fluorescent protein (EGFP). Here, we reprogrammed Oct4-EGFP fibroblasts employing the nonviral Sleeping Beauty transposon system to deliver the reprogramming factors Oct4, Sox2, Klf4, and cMyc. Successful reprogramming to a pluripotent state was indicated by changes in cell morphology and reactivation of the Oct4-EGFP reporter. The transposon-reprogrammed induced pluripotent stem (iPS) cells showed long-term proliferation in vitro over >40 passages, expressed transcription factors typical of embryonic stem cells, including OCT4, NANOG, SOX2, REX1, ESRRB, DPPA5, and UTF1 and surface markers of pluripotency, including SSEA-1 and TRA-1-60. In vitro differentiation resulted in derivatives of the 3 germ layers. Upon injection of putative iPS cells under the skin of immunodeficient mice, we observed teratomas in 3 of 6 cases. These results form the basis for in-depth studies toward the derivation of porcine iPS cells, which hold great promise for preclinical testing of novel cell therapies in the pig model.
Original languageEnglish
JournalStem Cells and Development
Volume22
Issue number1
Pages (from-to)124-135
Number of pages12
ISSN1547-3287
DOIs
Publication statusPublished - 2013

    Research areas

  • Animals, Antigens, Differentiation, Cell Transformation, Neoplastic, Cells, Cultured, Coculture Techniques, DNA Transposable Elements, Fibroblasts, Green Fluorescent Proteins, Induced Pluripotent Stem Cells, Mice, Mice, Nude, Microscopy, Fluorescence, Neurogenesis, Octamer Transcription Factor-3, Promoter Regions, Genetic, SOXB1 Transcription Factors, Sus scrofa, Teratoma, Transcriptome, Transgenes

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