Generation of induced pluripotent stem cells from human peripheral T cells using sendai virus in feeder-free conditions

Yoshikazu Kishino, Tomohisa Seki, Shinsuke Yuasa, Jun Fujita, Keiichi Fukuda

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Recently, iPSCs have attracted attention as a new source of cells for regenerative therapies. Although the initial method for generating iPSCs relied on dermal fibroblasts obtained by invasive biopsy and retroviral genomic insertion of transgenes, there have been many efforts to avoid these disadvantages. Human peripheral T cells are a unique cell source for generating iPSCs. iPSCs derived from T cells contain rearrangements of the T cell receptor (TCR) genes and are a source of antigen-specific T cells. Additionally, T cell receptor rearrangement in the genome has the potential to label individual cell lines and distinguish between transplanted and donor cells. For safe clinical application of iPSCs, it is important to minimize the risk of exposing newly generated iPSCs to harmful agents. Although fetal bovine serum and feeder cells have been essential for pluripotent stem cell culture, it is preferable to remove them from the culture system to reduce the risk of unpredictable pathogenicity. To address this, we have established a protocol for generating iPSCs from human peripheral T cells using Sendai virus to reduce the risk of exposing iPSCs to undefined pathogens. Although handling Sendai virus requires equipment with the appropriate biosafety level, Sendai virus infects activated T cells without genome insertion, yet with high efficiency. In this protocol, we demonstrate the generation of iPSCs from human peripheral T cells in feeder-free conditions using a combination of activated T cell culture and Sendai virus.

Original languageEnglish
Article numbere53225
JournalJournal of Visualized Experiments
Volume2015
Issue number105
DOIs
Publication statusPublished - 2015 Nov 11

Keywords

  • Cell reprogramming
  • Developmental biology
  • Induced pluripotent stem cells
  • Issue 105
  • Matrigel
  • Peripheral blood
  • Sendai virus
  • T cells

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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