Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions

Yoshikazu Kishino, Tomohisa Seki, Jun Fujita, Shinsuke Yuasa, Shugo Tohyama, Akira Kunitomi, Ryota Tabei, Kazuaki Nakajima, Marina Okada, Akinori Hirano, Hideaki Kanazawa, Keiichi Fukuda

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Recently, induced pluripotent stem cells (iPSCs) were established as promising cell sources for revolutionary regenerative therapies. The initial culture system used for iPSC generation needed fetal calf serum in the culture medium and mouse embryonic fibroblast as a feeder layer, both of which could possibly transfer unknown exogenous antigens and pathogens into the iPSC population. Therefore, the development of culture systems designed to minimize such potential risks has become increasingly vital for future applications of iPSCs for clinical use. On another front, although donor cell types for generating iPSCs are wide-ranging, T cells have attracted attention as unique cell sources for iPSCs generation because T cell-derived iPSCs (TiPSCs) have a unique monoclonal T cell receptor genomic rearrangement that enables their differentiation into antigen-specific T cells, which can be applied to novel immunotherapies. In the present study, we generated transgene-free human TiPSCs using a combination of activated human T cells and Sendai virus under defined culture conditions. These TiPSCs expressed pluripotent markers by quantitative PCR and immunostaining, had a normal karyotype, and were capable of differentiating into cells from all three germ layers. This method of TiPSCs generation is more suitable for the therapeutic application of iPSC technology because it lowers the risks associated with the presence of undefined, animal-derived feeder cells and serum. Therefore this work will lead to establishment of safer iPSCs and extended clinical application.

Original languageEnglish
Article numbere97397
JournalPLoS One
Volume9
Issue number5
DOIs
Publication statusPublished - 2014 May 13

Fingerprint

Induced Pluripotent Stem Cells
T-cells
Stem cells
Transgenes
Cell culture
transgenes
cell culture
T-lymphocytes
T-Lymphocytes
Feeder Cells
cells
differentiation antigens
Sendai virus
induced pluripotent stem cells
Antigens
Germ Layers
therapeutics
immunotherapy
Differentiation Antigens
fetal bovine serum

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions. / Kishino, Yoshikazu; Seki, Tomohisa; Fujita, Jun; Yuasa, Shinsuke; Tohyama, Shugo; Kunitomi, Akira; Tabei, Ryota; Nakajima, Kazuaki; Okada, Marina; Hirano, Akinori; Kanazawa, Hideaki; Fukuda, Keiichi.

In: PLoS One, Vol. 9, No. 5, e97397, 13.05.2014.

Research output: Contribution to journalArticle

Kishino, Yoshikazu ; Seki, Tomohisa ; Fujita, Jun ; Yuasa, Shinsuke ; Tohyama, Shugo ; Kunitomi, Akira ; Tabei, Ryota ; Nakajima, Kazuaki ; Okada, Marina ; Hirano, Akinori ; Kanazawa, Hideaki ; Fukuda, Keiichi. / Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions. In: PLoS One. 2014 ; Vol. 9, No. 5.
@article{5facc9d6e84f4848a1f67ba220ed4c40,
title = "Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions",
abstract = "Recently, induced pluripotent stem cells (iPSCs) were established as promising cell sources for revolutionary regenerative therapies. The initial culture system used for iPSC generation needed fetal calf serum in the culture medium and mouse embryonic fibroblast as a feeder layer, both of which could possibly transfer unknown exogenous antigens and pathogens into the iPSC population. Therefore, the development of culture systems designed to minimize such potential risks has become increasingly vital for future applications of iPSCs for clinical use. On another front, although donor cell types for generating iPSCs are wide-ranging, T cells have attracted attention as unique cell sources for iPSCs generation because T cell-derived iPSCs (TiPSCs) have a unique monoclonal T cell receptor genomic rearrangement that enables their differentiation into antigen-specific T cells, which can be applied to novel immunotherapies. In the present study, we generated transgene-free human TiPSCs using a combination of activated human T cells and Sendai virus under defined culture conditions. These TiPSCs expressed pluripotent markers by quantitative PCR and immunostaining, had a normal karyotype, and were capable of differentiating into cells from all three germ layers. This method of TiPSCs generation is more suitable for the therapeutic application of iPSC technology because it lowers the risks associated with the presence of undefined, animal-derived feeder cells and serum. Therefore this work will lead to establishment of safer iPSCs and extended clinical application.",
author = "Yoshikazu Kishino and Tomohisa Seki and Jun Fujita and Shinsuke Yuasa and Shugo Tohyama and Akira Kunitomi and Ryota Tabei and Kazuaki Nakajima and Marina Okada and Akinori Hirano and Hideaki Kanazawa and Keiichi Fukuda",
year = "2014",
month = "5",
day = "13",
doi = "10.1371/journal.pone.0097397",
language = "English",
volume = "9",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "5",

}

TY - JOUR

T1 - Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions

AU - Kishino, Yoshikazu

AU - Seki, Tomohisa

AU - Fujita, Jun

AU - Yuasa, Shinsuke

AU - Tohyama, Shugo

AU - Kunitomi, Akira

AU - Tabei, Ryota

AU - Nakajima, Kazuaki

AU - Okada, Marina

AU - Hirano, Akinori

AU - Kanazawa, Hideaki

AU - Fukuda, Keiichi

PY - 2014/5/13

Y1 - 2014/5/13

N2 - Recently, induced pluripotent stem cells (iPSCs) were established as promising cell sources for revolutionary regenerative therapies. The initial culture system used for iPSC generation needed fetal calf serum in the culture medium and mouse embryonic fibroblast as a feeder layer, both of which could possibly transfer unknown exogenous antigens and pathogens into the iPSC population. Therefore, the development of culture systems designed to minimize such potential risks has become increasingly vital for future applications of iPSCs for clinical use. On another front, although donor cell types for generating iPSCs are wide-ranging, T cells have attracted attention as unique cell sources for iPSCs generation because T cell-derived iPSCs (TiPSCs) have a unique monoclonal T cell receptor genomic rearrangement that enables their differentiation into antigen-specific T cells, which can be applied to novel immunotherapies. In the present study, we generated transgene-free human TiPSCs using a combination of activated human T cells and Sendai virus under defined culture conditions. These TiPSCs expressed pluripotent markers by quantitative PCR and immunostaining, had a normal karyotype, and were capable of differentiating into cells from all three germ layers. This method of TiPSCs generation is more suitable for the therapeutic application of iPSC technology because it lowers the risks associated with the presence of undefined, animal-derived feeder cells and serum. Therefore this work will lead to establishment of safer iPSCs and extended clinical application.

AB - Recently, induced pluripotent stem cells (iPSCs) were established as promising cell sources for revolutionary regenerative therapies. The initial culture system used for iPSC generation needed fetal calf serum in the culture medium and mouse embryonic fibroblast as a feeder layer, both of which could possibly transfer unknown exogenous antigens and pathogens into the iPSC population. Therefore, the development of culture systems designed to minimize such potential risks has become increasingly vital for future applications of iPSCs for clinical use. On another front, although donor cell types for generating iPSCs are wide-ranging, T cells have attracted attention as unique cell sources for iPSCs generation because T cell-derived iPSCs (TiPSCs) have a unique monoclonal T cell receptor genomic rearrangement that enables their differentiation into antigen-specific T cells, which can be applied to novel immunotherapies. In the present study, we generated transgene-free human TiPSCs using a combination of activated human T cells and Sendai virus under defined culture conditions. These TiPSCs expressed pluripotent markers by quantitative PCR and immunostaining, had a normal karyotype, and were capable of differentiating into cells from all three germ layers. This method of TiPSCs generation is more suitable for the therapeutic application of iPSC technology because it lowers the risks associated with the presence of undefined, animal-derived feeder cells and serum. Therefore this work will lead to establishment of safer iPSCs and extended clinical application.

UR - http://www.scopus.com/inward/record.url?scp=84901253207&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84901253207&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0097397

DO - 10.1371/journal.pone.0097397

M3 - Article

C2 - 24824994

AN - SCOPUS:84901253207

VL - 9

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 5

M1 - e97397

ER -