Efficient generation of integration-free human induced pluripotent stem cells from keratinocytes by simple transfection of episomal vectors

Yulan Piao, Sandy Shen Chi Hung, Shiang Y. Lim, Raymond Ching Bong Wong, Minoru S.H. Ko

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Keratinocytes represent an easily accessible cell source for derivation of human induced pluripotent stem (hiPS) cells, reportedly achieving higher reprogramming efficiency than fibroblasts. However, most studies utilized a retroviral or lentiviral method for reprogramming of keratinocytes, which introduces undesirable transgene integrations into the host genome. Moreover, current protocols of generating integration-free hiPS cells from keratinocytes are mostly inefficient. In this paper, we describe a more efficient, simple-to-use, and cost-effective method for generating integration-free hiPS cells from keratinocytes. Our improved method using lipid-mediated transfection achieved a reprogramming efficiency of ~0.14% on average. Keratinocyte-derived hiPS cells showed no integration of episomal vectors, expressed stem cell-specific markers and possessed potentials to differentiate into all three germ layers by in vitro embryoid body formation as well as in vivo teratoma formation. To our knowledge, this represents the most efficient method to generate integration-free hiPS cells from keratinocytes.

Original languageEnglish
Pages (from-to)787-791
Number of pages5
JournalStem Cells Translational Medicine
Volume3
Issue number7
DOIs
Publication statusPublished - 2014

Keywords

  • Derivation of human induced pluripotent stem cells
  • Episomal vectors
  • Integration-free
  • Keratinocytes
  • Lipid mediated transfection

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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