Skip to main content
SpringerLink
Log in
Book cover

p53 Protocols pp 157–164Cite as

Generation of p53-Deficient Induced Pluripotent Stem Cells from Mouse Embryo Fibroblasts

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 962))

Abstract

Here we describe a method for generating induced pluripotent stem (iPS) cells from mouse embryonic fibroblasts (MEFs). Recombinant retroviruses carrying human transcription factors for Klf4, Oct3/4, Sox2, with or without c-Myc, are used to transduce early passage MEFs several times. Based on morphologic criteria, the resulting iPS colonies are picked manually at first, and then propagated and expanded by standard methods. iPS cells can then be differentiated into virtually any cell type or lineage, thus allowing for discoveries of new functions of p53 and mutant p53.

This is a preview of subscription content, 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   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Okita K et al (2007) Generation of germline-competent induced pluripotent stem cells. Nature 448:313–317

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  3. Li H et al (2009) The Ink4/Arf locus is a barrier for iPS cell reprogramming. Nature 460:1136–1141

    Article  PubMed  CAS  Google Scholar 

  4. http://www.stanford.edu/group/nolan/retroviral_systems/phx.html

  5. https://www.wicell.org/index.php?option=com_docman%26task=doc_download%26gid=687

  6. Petrenko O et al (1999) The molecular characterization of the fetal stem cell marker AA4. Immunity 10:691–700

    Article  PubMed  CAS  Google Scholar 

  7. http://www.stemcell.com/~/media/Tech­nical%20Resources/2/29141MAN_1_1_1.ashx

  8. Whibley C et al (2010) Wild-type and Hupki (human p53 knock-in) murine embryonic fibroblasts: p53/ARF pathway disruption in spontaneous escape from senescence. J Biol Chem 285:11326–11335

    Article  PubMed  CAS  Google Scholar 

  9. Hong H et al (2009) Suppression of induced pluripotent stem cell generation by the p53-p21 pathway. Nature 460:1132–1135

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Dr. Olexi Petrenko for sharing his expertise and generating pRebna constructs of reprogramming factors, Dr. Alice Nemajerova for technical advice and useful discussions, and Dr. Flaminia Talos for critical reading of this chapter.

Author information

Authors and Affiliations

  1. Department of Pathology, Stony Brook University, Stony Brook, NY, USA

    Evguenia M. Alexandrova & Ute M. Moll

Authors
  1. Evguenia M. Alexandrova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ute M. Moll
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ute M. Moll .

Editor information

Editors and Affiliations

  1. Massey Cancer Center, Department of Biochemistry, Virginia Commonwealth University, College Street 401, Richmond, 23298, Virginia, USA

    Sumitra Deb

  2. Massey Cancer Center, Department of Biochemistry, Virginia Commonwealth University, College Street 401, Richmond, 23298, Virginia, USA

    Swati Palit Deb

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media New York

About this protocol

Cite this protocol

Alexandrova, E.M., Moll, U.M. (2013). Generation of p53-Deficient Induced Pluripotent Stem Cells from Mouse Embryo Fibroblasts. In: Deb, S., Deb, S. (eds) p53 Protocols. Methods in Molecular Biology, vol 962. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-236-0_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-236-0_13

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-235-3

  • Online ISBN: 978-1-62703-236-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation