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


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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: KiPS cell colony formation and cell line characterization.
Figure 2: KiPS cells display retroviral silencing, promoter demethylation and activation of endogenous pluripotency factors.
Figure 5: q-PCR and microarray analysis comparing keratinocytes, fibroblasts, KiPS cells and ES cells.
Figure 3: KiPS cells can differentiate into all three primary germ layers in vitro and in vivo.
Figure 4: The dynamics of efficient keratinocyte reprogramming.
Figure 6: Generation and characterization of KiPS cells from a single plucked hair.

Accession codes


Gene Expression Omnibus


  1. 1

    Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006).

    CAS  Article  Google Scholar 

  2. 2

    Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007).

    CAS  Article  Google Scholar 

  3. 3

    Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917–1920 (2007).

    CAS  Article  Google Scholar 

  4. 4

    Lowry, W.E. et al. Generation of human induced pluripotent stem cells from dermal fibroblasts. Proc. Natl. Acad. Sci. USA 105, 2883–2888 (2008).

    CAS  Article  Google Scholar 

  5. 5

    Park, I.H. et al. Reprogramming of human somatic cells to pluripotency with defined factors. Nature 451, 141–146 (2008).

    CAS  Article  Google Scholar 

  6. 6

    Aoi, T. et al. Generation of pluripotent stem cells from adult mouse liver and stomach cells. Science 321, 699–702 (2008).

    CAS  Article  Google Scholar 

  7. 7

    Wernig, M. et al. A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types. Nat. Biotechnol. 26, 916–924 (2008).

    CAS  Article  Google Scholar 

  8. 8

    Fuchs, E. Scratching the surface of skin development. Nature 445, 834–842 (2007).

    CAS  Article  Google Scholar 

  9. 9

    Unsworth, H.C., Aasen, T., McElwaine, S. & Kelsell, D.P. Tissue-specific effects of wild-type and mutant connexin 31: a role in neurite outgrowth. Hum. Mol. Genet. 16, 165–172 (2007).

    CAS  Article  Google Scholar 

  10. 10

    Hawley, R.G., Lieu, F.H., Fong, A.Z. & Hawley, T.S. Versatile retroviral vectors for potential use in gene therapy. Gene Ther. 1, 136–138 (1994).

    CAS  PubMed  Google Scholar 

  11. 11

    Nakagawa, M. et al. Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat. Biotechnol. 26, 101–106 (2008).

    CAS  Article  Google Scholar 

  12. 12

    O'Connor, M.D. et al. Alkaline phosphatase-positive colony formation is a sensitive, specific, and quantitative indicator of undifferentiated human embryonic stem cells. Stem Cells 26, 1109–1116 (2008).

    CAS  Article  Google Scholar 

  13. 13

    Draper, J.S. et al. Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat. Biotechnol. 22, 53–54 (2004).

    CAS  Article  Google Scholar 

  14. 14

    Mitalipova, M.M. et al. Preserving the genetic integrity of human embryonic stem cells. Nat. Biotechnol. 23, 19–20 (2005).

    CAS  Article  Google Scholar 

  15. 15

    Raya, A. et al. Generation of cardiomyocytes from new human embryonic stem cell lines derived from poor-quality blastocysts. Cold Spring Harb. Symp. Quant. Biol. (in press).

  16. 16

    Brambrink, T. et al. Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells. Cell Stem Cell 2, 151–159 (2008).

    CAS  Article  Google Scholar 

  17. 17

    Assou, S. et al. A meta-analysis of human embryonic stem cells transcriptome integrated into a web-based expression atlas. Stem Cells 25, 961–973 (2007).

    CAS  Article  Google Scholar 

  18. 18

    Limat, A. & Noser, F.K. Serial cultivation of single keratinocytes from the outer root sheath of human scalp hair follicles. J. Invest. Dermatol. 87, 485–488 (1986).

    CAS  Article  Google Scholar 

  19. 19

    Kurata, S., Itami, S., Terashi, H. & Takayasu, S. Successful transplantation of cultured human outer root sheath cells as epithelium. Ann. Plast. Surg. 33, 290–294 (1994).

    CAS  Article  Google Scholar 

  20. 20

    Sridharan, R. & Plath, K. Illuminating the black box of reprogramming. Cell Stem Cell 2, 295–297 (2008).

    CAS  Article  Google Scholar 

  21. 21

    Segre, J.A., Bauer, C. & Fuchs, E. Klf4 is a transcription factor required for establishing the barrier function of the skin. Nat. Genet. 22, 356–360 (1999).

    CAS  Article  Google Scholar 

  22. 22

    Gandarillas, A. & Watt, F.M. c-Myc promotes differentiation of human epidermal stem cells. Genes Dev. 11, 2869–2882 (1997).

    CAS  Article  Google Scholar 

  23. 23

    Kim, J.B. et al. Pluripotent stem cells induced from adult neural stem cells by reprogramming with two factors. Nature 454, 646–650 (2008).

    CAS  Article  Google Scholar 

  24. 24

    Amoh, Y., Li, L., Katsuoka, K., Penman, S. & Hoffman, R.M. Multipotent nestin-positive, keratin-negative hair-follicle bulge stem cells can form neurons. Proc. Natl. Acad. Sci. USA 102, 5530–5534 (2005).

    CAS  Article  Google Scholar 

  25. 25

    Yu, H. et al. Isolation of a novel population of multipotent adult stem cells from human hair follicles. Am. J. Pathol. 168, 1879–1888 (2006).

    CAS  Article  Google Scholar 

  26. 26

    Reich, M. et al. GenePattern 2.0. Nat. Genet. 38, 500–501 (2006).

    CAS  Article  Google Scholar 

  27. 27

    de Hoon, M.J., Imoto, S., Nolan, J. & Miyano, S. Open source clustering software. Bioinformatics 20, 1453–1454 (2004).

    CAS  Article  Google Scholar 

  28. 28

    Freberg, C.T., Dahl, J.A., Timoskainen, S. & Collas, P. Epigenetic reprogramming of OCT4 and NANOG regulatory regions by embryonal carcinoma cell extract. Mol. Biol. Cell 18, 1543–1553 (2007).

    CAS  Article  Google Scholar 

  29. 29

    Virtaneva, K. et al. Expression profiling reveals fundamental biological differences in acute myeloid leukemia with isolated trisomy 8 and normal cytogenetics. Proc. Natl. Acad. Sci. USA 98, 1124–1129 (2001).

    CAS  Article  Google Scholar 

  30. 30

    Wu, Z., Irizarry, R.A., Gentleman, R., Martinez-Murillo, F. & Spencer, F. A model-based background adjustment for oligonucleotide expression arrays. J. Am. Stat. Assoc. 99, 909–917 (2004).

    Article  Google Scholar 

Download references


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 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.

Author information



Corresponding author

Correspondence to Juan Carlos Izpisúa Belmonte.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–13, Supplementary Data (PDF 24671 kb)

Supplementary Movie 1

Real-time movie of KiPS4F4 cells differentiated into beating cardiomyocytes. (MOV 3384 kb)

Supplementary Movie 2

Real-time movie of hair-derived iPS cells differentiated into beating cardiomyocytes (Hair sample 1). (MOV 2843 kb)

Supplementary Movie 3

Real-time movie of hair-derived iPS cells differentiated into beating cardiomyocytes (Hair sample 2). (MOV 2678 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Aasen, T., Raya, A., Barrero, M. et al. Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes. Nat Biotechnol 26, 1276–1284 (2008). https://doi.org/10.1038/nbt.1503

Download citation

Further reading


Sign up for the Nature Briefing newsletter for a daily update on COVID-19 science.
Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing