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

Fingerprint

Sendai virus
Induced Pluripotent Stem Cells
T-cells
Stem cells
Viruses
T-Lymphocytes
Genes
T-Cell Antigen Receptor
Cell culture
Cell Culture Techniques
Genome
Feeder Cells
T-Cell Receptor Genes
Pluripotent Stem Cells
Biopsy
Pathogens
Fibroblasts
Cell- and Tissue-Based Therapy
Transgenes
Virulence

Keywords

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

ASJC Scopus subject areas

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

Cite this

Generation of induced pluripotent stem cells from human peripheral T cells using sendai virus in feeder-free conditions. / Kishino, Yoshikazu; Seki, Tomohisa; Yuasa, Shinsuke; Fujita, Jun; Fukuda, Keiichi.

In: Journal of Visualized Experiments, Vol. 2015, No. 105, e53225, 11.11.2015.

Research output: Contribution to journalArticle

@article{e1974375e4ca4fa78533bc7e7157e1de,
title = "Generation of induced pluripotent stem cells from human peripheral T cells using sendai virus in feeder-free conditions",
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.",
keywords = "Cell reprogramming, Developmental biology, Induced pluripotent stem cells, Issue 105, Matrigel, Peripheral blood, Sendai virus, T cells",
author = "Yoshikazu Kishino and Tomohisa Seki and Shinsuke Yuasa and Jun Fujita and Keiichi Fukuda",
year = "2015",
month = "11",
day = "11",
doi = "10.3791/53225",
language = "English",
volume = "2015",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "105",

}

TY - JOUR

T1 - Generation of induced pluripotent stem cells from human peripheral T cells using sendai virus in feeder-free conditions

AU - Kishino, Yoshikazu

AU - Seki, Tomohisa

AU - Yuasa, Shinsuke

AU - Fujita, Jun

AU - Fukuda, Keiichi

PY - 2015/11/11

Y1 - 2015/11/11

N2 - 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.

AB - 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.

KW - Cell reprogramming

KW - Developmental biology

KW - Induced pluripotent stem cells

KW - Issue 105

KW - Matrigel

KW - Peripheral blood

KW - Sendai virus

KW - T cells

UR - http://www.scopus.com/inward/record.url?scp=84949472605&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84949472605&partnerID=8YFLogxK

U2 - 10.3791/53225

DO - 10.3791/53225

M3 - Article

AN - SCOPUS:84949472605

VL - 2015

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 105

M1 - e53225

ER -