Generation of induced pluripotent stem cells with CytoTune, a non-integrating Sendai virus

Methods Mol Biol. 2013:997:45-56. doi: 10.1007/978-1-62703-348-0_5.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / metabolism
  • Base Sequence
  • Cell Dedifferentiation
  • Coculture Techniques
  • DNA Primers / genetics
  • Feeder Cells
  • Fibroblasts / physiology
  • Genetic Vectors
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / physiology*
  • Matrix Metalloproteinase 2 / chemistry
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sendai virus / genetics*
  • Transcription Factors / genetics

Substances

  • Antigens, CD34
  • DNA Primers
  • Transcription Factors
  • Matrix Metalloproteinase 2