Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes

Trond Aasen; Angel Raya; Maria J. Barrero; Elena Garreta; Antonella Consiglio; Federico Gonzalez; Rita Vassena; Josipa Bilić; Vladimir Pekarik; Gustavo Tiscornia; Michael Edel; Stéphanie Boué; Juan Carlos Izpisúa Belmonte

doi:10.1038/nbt.1503

Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes

Trond Aasen, Angel Raya, Maria J. Barrero, Elena Garreta, Antonella Consiglio, Federico Gonzalez, Rita Vassena, Josipa Bilić, Vladimir Pekarik, Gustavo Tiscornia, Michael Edel, Stéphanie Boué, Juan Carlos Izpisúa Belmonte

Research output: Contribution to journal › Article › peer-review

1185 Scopus citations

Abstract

The utility of induced pluripotent stem (iPS) cells for investigating the molecular logic of pluripotency and for eventual clinical application is limited by the low efficiency of current methods for reprogramming. Here we show that reprogramming of juvenile human primary keratinocytes by retroviral transduction with OCT4, SOX2, KLF4 and c-MYC is at least 100-fold more efficient and twofold faster compared with reprogramming of human fibroblasts. Keratinocyte-derived iPS (KiPS) cells appear indistinguishable from human embryonic stem cells in colony morphology, growth properties, expression of pluripotency-associated transcription factors and surface markers, global gene expression profiles and differentiation potential in vitro and in vivo. To underscore the efficiency and practicability of this technology, we generated KiPS cells from single adult human hairs. Our findings provide an experimental model for investigating the bases of cellular reprogramming and highlight potential advantages of using keratinocytes to generate patient-specific iPS cells.

Original language	English (US)
Pages (from-to)	1276-1284
Number of pages	9
Journal	Nature biotechnology
Volume	26
Issue number	11
DOIs	https://doi.org/10.1038/nbt.1503
State	Published - Nov 2008
Externally published	Yes

Bibliographical note

Funding Information:
We are grateful to Ignacio Pizá Rodriguez for help and advice with KiPS cell characterization; José Miguel Andrés Vaquero for assistance with FACS analysis; Meritxell Carrió for expert assistance with cell culture techniques; Esther Melo, Lola Mulero Pérez and Mercé Gaudes Martí for bioimaging assistance; Yvonne Richaud and Teresa Lopez Rovira for excellent technical assistance and Luciano Di Croce, Centre for Genomic Regulation, Barcelona, for the gift of c-MYC T58A plasmid. F.G. was partially supported by a fellowship from the Swiss National Science Foundation. M.J.B. and G.T. were partially supported by the Ramón y Cajal program. This work was partially supported by grants from Ministerio de

Funding Information:
Educación y Ciencia grant BFU2006-12251, European Commission ‘Marie-Curie Reintegration Grant’ MIRG-CT-2007-046523 the Fondo de Investigaciones Sanitarias (RETIC-RD06/0010/0016, PI061897), Marató de TV3 (063430), the G. Harold and Leila Y. Mathers Charitable Foundation and Fundación Cellex.

ASJC Scopus subject areas

Applied Microbiology and Biotechnology
Bioengineering
Molecular Medicine
Biotechnology
Biomedical Engineering

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10.1038/nbt.1503

Cite this

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title = "Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes",

abstract = "The utility of induced pluripotent stem (iPS) cells for investigating the molecular logic of pluripotency and for eventual clinical application is limited by the low efficiency of current methods for reprogramming. Here we show that reprogramming of juvenile human primary keratinocytes by retroviral transduction with OCT4, SOX2, KLF4 and c-MYC is at least 100-fold more efficient and twofold faster compared with reprogramming of human fibroblasts. Keratinocyte-derived iPS (KiPS) cells appear indistinguishable from human embryonic stem cells in colony morphology, growth properties, expression of pluripotency-associated transcription factors and surface markers, global gene expression profiles and differentiation potential in vitro and in vivo. To underscore the efficiency and practicability of this technology, we generated KiPS cells from single adult human hairs. Our findings provide an experimental model for investigating the bases of cellular reprogramming and highlight potential advantages of using keratinocytes to generate patient-specific iPS cells.",

author = "Trond Aasen and Angel Raya and Barrero, {Maria J.} and Elena Garreta and Antonella Consiglio and Federico Gonzalez and Rita Vassena and Josipa Bili{\'c} and Vladimir Pekarik and Gustavo Tiscornia and Michael Edel and St{\'e}phanie Bou{\'e} and Belmonte, {Juan Carlos Izpis{\'u}a}",

note = "Funding Information: We are grateful to Ignacio Piz{\'a} Rodriguez for help and advice with KiPS cell characterization; Jos{\'e} Miguel Andr{\'e}s Vaquero for assistance with FACS analysis; Meritxell Carri{\'o} for expert assistance with cell culture techniques; Esther Melo, Lola Mulero P{\'e}rez and Merc{\'e} Gaudes Mart{\'i} for bioimaging assistance; Yvonne Richaud and Teresa Lopez Rovira for excellent technical assistance and Luciano Di Croce, Centre for Genomic Regulation, Barcelona, for the gift of c-MYC T58A plasmid. F.G. was partially supported by a fellowship from the Swiss National Science Foundation. M.J.B. and G.T. were partially supported by the Ram{\'o}n y Cajal program. This work was partially supported by grants from Ministerio de Funding Information: Educaci{\'o}n y Ciencia grant BFU2006-12251, European Commission {\textquoteleft}Marie-Curie Reintegration Grant{\textquoteright} MIRG-CT-2007-046523 the Fondo de Investigaciones Sanitarias (RETIC-RD06/0010/0016, PI061897), Marat{\'o} de TV3 (063430), the G. Harold and Leila Y. Mathers Charitable Foundation and Fundaci{\'o}n Cellex.",

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AU - Gonzalez, Federico

AU - Vassena, Rita

AU - Bilić, Josipa

AU - Pekarik, Vladimir

AU - Tiscornia, Gustavo

AU - Edel, Michael

AU - Boué, Stéphanie

AU - Belmonte, Juan Carlos Izpisúa

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Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes

Abstract

Bibliographical note

ASJC Scopus subject areas

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