Skip to main content

Generation of Induced Pluripotent Stem Cells with CytoTune, a Non-Integrating Sendai Virus

  • Protocol
  • First Online:
Pluripotent Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 997))

Abstract

One of the major obstacles in generating induced pluripotent stem cells for research or downstream applications is the potential modifications of cellular genome as a result of using integrating viruses during reprogramming. Another major disadvantage of reprogramming cells with integrating vectors is that silencing and activation of transgenes are unpredictable, which may affect terminal differentiation potential and increase the risk of using iPSC-derived cells. Here we describe a protocol for the generation of induced pluripotent stem cells using a non-integrating RNA virus, Sendai virus, to efficiently generate transgene-free iPSCs starting with different cell types as well as in feeder-free conditions.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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

References

  1. Takeda A, Igarashi H, Kawada M, Tsukamoto T, Yamamoto H, Inoue M, Iida A, Shu T, Hasegawa M, Matano T (2008) Evaluation of the immunogenicity of replication-competent V-knocked-out and replication-defective F-deleted Sendai virus vector-based vaccines in macaques. Vaccine 26:6839–6843

    Article  PubMed  CAS  Google Scholar 

  2. Wernig M, Meissner A, Foreman R, Brambrink T, Ku M, Hochedlinger K, Bernstein BE, Jaenisch R (2007) In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 448:318–324

    Article  PubMed  CAS  Google Scholar 

  3. Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science 318:1917–1920

    Article  PubMed  CAS  Google Scholar 

  4. Park IH, Lerou PH, Zhao R, Huo H, Daley GQ (2008) Generation of human-induced pluripotent stem cells. Nat Protoc 3: 1180–1186

    Article  PubMed  CAS  Google Scholar 

  5. Dimos JT, Rodolfa KT, Niakan KK, Weisenthal LM, Mitsumoto H, Chung W, Croft GF, Saphier G, Leibel R, Goland R, Wichterle H, Henderson CE, Eggan K (2008) Induced pluripotent stem cells generated from patients with ALS can be differentiated into motor neurons. Science 321:1218–1221

    Article  PubMed  CAS  Google Scholar 

  6. Ebert AD, Yu J, Rose FF Jr, Mattis VB, Lorson CL, Thomson JA, Svendsen CN (2009) Induced pluripotent stem cells from a spinal muscular atrophy patient. Nature 457: 277–280

    Article  PubMed  CAS  Google Scholar 

  7. Soldner F, Hockemeyer D, Beard C, Gao Q, Bell GW, Cook EG, Hargus G, Blak A, Cooper O, Mitalipova M, Isacson O, Jaenisch R (2009) Parkinson’s disease patient-derived induced pluripotent stem cells free of viral reprogramming factors. Cell 136:964–977

    Article  PubMed  CAS  Google Scholar 

  8. Okita K, Ichisaka T, Yamanaka S (2007) Generation of germline-competent induced pluripotent stem cells. Nature 448:313–317

    Article  PubMed  CAS  Google Scholar 

  9. Okita K, Matsumura Y, Sato Y, Okada A, Morizane A, Okamoto S, Hong H, Nakagawa M, Tanabe K, Tezuka K, Shibata T, Kunisada T, Takahashi M, Takahashi J, Saji H, Yamanaka S (2011) A more efficient method to generate integration-free human iPS cells. Nat Methods 8:409–412

    Article  PubMed  CAS  Google Scholar 

  10. Muller LU, Daley GQ, Williams DA (2009) Upping the ante: recent advances in direct reprogramming. Mol Ther 17:947–953

    Article  PubMed  CAS  Google Scholar 

  11. Li HO, Zhu YF, Asakawa M, Kuma H, Hirata T, Ueda Y, Lee YS, Fukumura M, Iida A, Kato A, Nagai Y, Hasegawa M (2000) A cytoplasmic RNA vector derived from nontransmissible Sendai virus with efficient gene transfer and expression. J Virol 74:6564–6569

    Article  PubMed  CAS  Google Scholar 

  12. Ikeda Y, Yonemitsu Y, Sakamoto T, Ishibashi T, Ueno H, Kato A, Nagai Y, Fukumura M, Inomata H, Hasegawa M, Sueishi K (2002) Recombinant Sendai virus-mediated gene transfer into adult rat retinal tissue: efficient gene transfer by brief exposure. Exp Eye Res 75:39–48

    Article  PubMed  CAS  Google Scholar 

  13. Masaki I, Yonemitsu Y, Komori K, Ueno H, Nakashima Y, Nakagawa K, Fukumura M, Kato A, Hasan MK, Nagai Y, Sugimachi K, Hasegawa M, Sueishi K (2001) Recombinant Sendai virus-mediated gene transfer to ­vasculature: a new class of efficient gene transfer vector to the vascular system. FASEB J 15:1294–1296

    PubMed  CAS  Google Scholar 

  14. Fusaki N, Ban H, Nishiyama A, Saeki K, Hasegawa M (2009) Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome. Proc Jpn Acad 85:348–362

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by Life Technologies Corporation. The products within this publication are for Research Use Only, not intended for animal or human therapeutic or diagnostic use.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media New York

About this protocol

Cite this protocol

Lieu, P.T., Fontes, A., Vemuri, M.C., MacArthur, C.C. (2013). Generation of Induced Pluripotent Stem Cells with CytoTune, a Non-Integrating Sendai Virus. In: Lakshmipathy, U., Vemuri, M. (eds) Pluripotent Stem Cells. Methods in Molecular Biology, vol 997. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-348-0_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-348-0_5

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-347-3

  • Online ISBN: 978-1-62703-348-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics