Using the Inducible Caspase-9 Suicide-Safeguard System with iPSC and Bioluminescent Tracking

  • Jonathan Villanueva
  • Toshinobu Nishimura
  • Hiromitsu NakauchiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2048)


For scientists working within the field of induced pluripotent stem cells (iPSCs), this protocol will provide a thorough walk-through on how to conduct in vitro and in vivo experiments that validate the function of a particular safeguard system technology. In short, we provide instructions on how to generate inducible Caspase-9 (iC9) safeguard system with human iPSCs that act as normal or abnormal models of the cells for therapeutics to be tried after differentiation. These iC9-iPSCs should be modified prior to beginning this protocol by constitutively expressing luciferase, an enzyme capable of generating bioluminescent signals through the oxidation of the substrate luciferin. Monitoring the bioluminescent signal over time provides the information on whether a safeguard system is working or not.

Key words

Immunotherapy Cancer Bioluminescence Cytotoxicity T cell 


  1. 1.
    Fesnak AD, June CH, Levine BL (2016) Engineered T cells: the promise and challenges of cancer immunotherapy. Nat Rev Cancer 16:566–581. Scholar
  2. 2.
    Zhao S, Weinreich MA, Ihara K, Richard RE, Blau CA (2004) In vivo selection of genetically modified erythroid cells using a jak2-based cell growth switch. Mol Ther 10:456–468. Scholar
  3. 3.
    Peterson SE et al (2011) Teratoma generation in the testis capsule. J Vis Exp (57):e3177.
  4. 4.
    Ando M et al (2015) A safeguard system for induced pluripotent stem cell-derived rejuvenated T cell therapy. Stem Cell Reports 5:597–608. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jonathan Villanueva
    • 1
  • Toshinobu Nishimura
    • 1
  • Hiromitsu Nakauchi
    • 1
    Email author
  1. 1.Institute for Stem Cell Biology and Regenerative MedicineStanford University School of MedicineStanfordUSA

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