Skip to main content
SpringerLink
Log in

Pluripotent Stem Cell Differentiation Toward Functional Basal Stratified Epithelial Cells

  • Protocol
  • First Online:
Induced Pluripotent Stem (iPS) Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2454))

Abstract

In this chapter, an efficient feeder-free protocol of differentiating human pluripotent stem cells (hPSCs) toward the epidermal lineage to generate induced epidermal keratinocytes (iKCs) is described. The iKCs are able to terminally differentiate supra-basally. This hPSC-to-iKC differentiation can serve as a useful model to study epidermal development and disease as well as for therapeutic applications.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 279.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Takahashi K, Yamanaka S (2016) A decade of transcription factor-mediated reprogramming to pluripotency. Nat Rev Mol Cell Biol 17:183–193. https://doi.org/10.1038/nrm.2016.8

    Article  CAS  PubMed  Google Scholar 

  2. Soares E, Xu Q, Li Q et al (2019) Single-cell RNA-seq identifies a reversible mesenchymal activation in abnormally specified epithelia of p63 EEC syndrome. Proc Natl Acad Sci U S A 116:437632. https://doi.org/10.1101/437632

    Article  CAS  Google Scholar 

  3. Shalom-Feuerstein R, Serror L, Aberdam E et al (2013) Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1MET. Proc Natl Acad Sci U S A 110:2152–2156. https://doi.org/10.1073/pnas.1201753109

    Article  PubMed  PubMed Central  Google Scholar 

  4. Shen J, van den Bogaard EH, Kouwenhoven EN et al (2013) APR-246/PRIMA-1MET rescues epidermal differentiation in skin keratinocytes derived from EEC syndrome patients with p63 mutations. Proc Natl Acad Sci 110:2157–2162. https://doi.org/10.1073/pnas.1201993110

    Article  PubMed  PubMed Central  Google Scholar 

  5. Umegaki-Arao N, Pasmooij AMG, Itoh M et al (2014) Induced pluripotent stem cells from human revertant keratinocytes for the treatment of epidermolysis bullosa. Sci Transl Med 6:1–10. https://doi.org/10.1126/scitranslmed.3009342

    Article  CAS  Google Scholar 

  6. Sebastiano V, Zhen HH, Haddad B et al (2014) Human COL7A1-corrected induced pluripotent stem cells for the treatment of recessive dystrophic epidermolysis bullosa. Sci Transl Med 6:1–12

    Article  Google Scholar 

  7. Ji LIN, Allen-hoffmann BL, Ph D et al (2006) Generation and differentiation of human embryonic stem cell-derived keratinocyte precursors. Tissue Eng 12:665–679

    Article  CAS  Google Scholar 

  8. Schuldiner M, Yanuka O, Itskovitz-eldor J et al (2000) Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells. Proc Natl Acad Sci U S A 97:11307–11312

    Article  CAS  Google Scholar 

  9. Bagutti C, Hutter C, Chiquet-ehrismann R, Watt FM (2001) Dermal fibroblast-derived growth factors restore the ability of Beta 1 integrin-deficient Embryonal stem cells to differentiate into keratinocytes. Dev Biol 333:321–333. https://doi.org/10.1006/dbio.2000.0149

    Article  CAS  Google Scholar 

  10. Coraux C, Hilmi C, Rouleau M et al (2003) Reconstituted skin from murine embryonic stem cells. Curr Biol 13:849–853. https://doi.org/10.1016/s0960-9822(03)00296-3

    Article  CAS  PubMed  Google Scholar 

  11. Troy T-C, Turksen K (2005) Commitment of embryonic stem cells to an epidermal cell fate and differentiation in vitro. Dev Dyn 232:293–300. https://doi.org/10.1002/dvdy.20223

    Article  CAS  PubMed  Google Scholar 

  12. Itoh M, Umegaki-Arao N, Guo Z et al (2013) Generation of 3D skin equivalents fully reconstituted from human induced pluripotent stem cells (iPSCs). PLoS One 8:1–9. https://doi.org/10.1371/journal.pone.0077673

    Article  CAS  Google Scholar 

  13. Bilousova G, Chen J, Roop DR (2011) Differentiation of mouse induced pluripotent stem cells into a multipotent keratinocyte lineage. J Invest Dermatol 131:857–864. https://doi.org/10.1038/jid.2010.364

    Article  CAS  PubMed  Google Scholar 

  14. Pattison JM, Melo SP, Piekos SN et al (2018) Retinoic acid and BMP4 cooperate with p63 to alter chromatin dynamics during surface epithelial commitment. Nat Genet 50:1658–1665. https://doi.org/10.1038/s41588-018-0263-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kogut I, Roop DR, Bilousova G (2014) Differentiation of human induced pluripotent stem cells into a keratinocyte lineage. Methods Mol Biol 1195:1–12. https://doi.org/10.1007/7651_2013_64

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Li L, Wang Y, Torkelson JL et al (2019) TFAP2C- and p63-dependent networks sequentially rearrange chromatin landscapes to drive human epidermal lineage commitment. Cell Stem Cell 24:271–284.e8. https://doi.org/10.1016/j.stem.2018.12.012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by Royal Dutch Academy of Science (KNAW/CEP 530-5CDP08) and Brazilian Science without Borders program (ES).

Author information

Authors and Affiliations

  1. Department of Molecular Developmental Biology, Faculty of Science, Radboud Institute for Molecular Life Sciences, Radboud University, Nijmegen, The Netherlands

    Eduardo Soares & Huiqing Zhou

  2. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Foundation, Ministry of Education of Brazil, Brasília, Brazil

    Eduardo Soares

  3. Department of Human Genetics, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands

    Huiqing Zhou

Authors
  1. Eduardo Soares
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Huiqing Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Huiqing Zhou .

Editor information

Editors and Affiliations

  1. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada

    Andras Nagy

  2. Ottawa, ON, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media New York

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Soares, E., Zhou, H. (2020). Pluripotent Stem Cell Differentiation Toward Functional Basal Stratified Epithelial Cells. In: Nagy, A., Turksen, K. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 2454. Humana, New York, NY. https://doi.org/10.1007/7651_2020_301

Download citation

  • DOI: https://doi.org/10.1007/7651_2020_301

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2118-9

  • Online ISBN: 978-1-0716-2119-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation