Removal of reprogramming transgenes improves the tissue reconstitution potential of keratinocytes generated from human induced pluripotent stem cells

Ken Igawa, Chikara Kokubu, Kosuke Yusa, Kyoji Horie, Yasuhide Yoshimura, Kaori Yamauchi, Hirofumi Suemori, Hiroo Yokozeki, Masashi Toyoda, Nobutaka Kiyokawa, Hajime Okita, Yoshitaka Miyagawa, Hidenori Akutsu, Akihiro Umezawa, Ichiro Katayama, Junji Takeda

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Human induced pluripotent stem cell (hiPSC) lines have a great potential for therapeutics because customized cells and organs can be induced from such cells. Assessment of the residual reprogramming factors after the generation of hiPSC lines is required, but an ideal system has been lacking. Here, we generated hiPSC lines from normal human dermal fibroblasts with piggyBac transposon bearing reprogramming transgenes followed by removal of the transposon by the transposase. Under this condition, we compared the phenotypes of transgene-residual and -free hiPSCs of the same genetic background. The transgene-residual hiPSCs, in which the transcription levels of the reprogramming transgenes were eventually suppressed, were quite similar to the transgene-free hiPSCs in a pluripotent state. However, after differentiation into keratinocytes, clear differences were observed. Morphological, functional, and molecular analyses including single-cell gene expression profiling revealed that keratinocytes from transgene-free hiPSC lines were more similar to normal human keratinocytes than those from transgene-residual hiPSC lines, which may be partly explained by reactivation of residual transgenes upon induction of keratinocyte differentiation. These results suggest that transgene-free hiPSC lines should be chosen for therapeutic purposes.

Original languageEnglish
Pages (from-to)992-1001
Number of pages10
JournalStem Cells Translational Medicine
Volume3
Issue number9
DOIs
Publication statusPublished - 2014
Externally publishedYes

Keywords

  • Differentiation
  • Human
  • Induced pluripotent stem cell
  • Keratinocyte
  • Transgene

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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  • Cite this

    Igawa, K., Kokubu, C., Yusa, K., Horie, K., Yoshimura, Y., Yamauchi, K., Suemori, H., Yokozeki, H., Toyoda, M., Kiyokawa, N., Okita, H., Miyagawa, Y., Akutsu, H., Umezawa, A., Katayama, I., & Takeda, J. (2014). Removal of reprogramming transgenes improves the tissue reconstitution potential of keratinocytes generated from human induced pluripotent stem cells. Stem Cells Translational Medicine, 3(9), 992-1001. https://doi.org/10.5966/sctm.2013-0179