TY - JOUR
T1 - Removal of reprogramming transgenes improves the tissue reconstitution potential of keratinocytes generated from human induced pluripotent stem cells
AU - Igawa, Ken
AU - Kokubu, Chikara
AU - Yusa, Kosuke
AU - Horie, Kyoji
AU - Yoshimura, Yasuhide
AU - Yamauchi, Kaori
AU - Suemori, Hirofumi
AU - Yokozeki, Hiroo
AU - Toyoda, Masashi
AU - Kiyokawa, Nobutaka
AU - Okita, Hajime
AU - Miyagawa, Yoshitaka
AU - Akutsu, Hidenori
AU - Umezawa, Akihiro
AU - Katayama, Ichiro
AU - Takeda, Junji
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - Differentiation
KW - Human
KW - Induced pluripotent stem cell
KW - Keratinocyte
KW - Transgene
UR - http://www.scopus.com/inward/record.url?scp=84906846740&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84906846740&partnerID=8YFLogxK
U2 - 10.5966/sctm.2013-0179
DO - 10.5966/sctm.2013-0179
M3 - Article
C2 - 25024429
AN - SCOPUS:84906846740
VL - 3
SP - 992
EP - 1001
JO - Stem cells translational medicine
JF - Stem cells translational medicine
SN - 2157-6564
IS - 9
ER -