Protocol

Pluripotent Stem Cells

Volume 997 of the series Methods in Molecular Biology pp 45-56

Date:

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

  • Pauline T. LieuAffiliated withPrimary and Stem Cell Systems, Life Technologies
  • , Andrew FontesAffiliated withPrimary and Stem Cell Systems, Life Technologies
  • , Mohan C. VemuriAffiliated withPrimary and Stem Cell Systems, Life Technologies
  • , Chad C. MacArthurAffiliated withPrimary and Stem Cell Systems, Life Technologies

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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.

Key words

Induced pluripotent stem cells Non-integrating Sendai virus High efficiency Human cells Reprogramming Feeder free