Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions.
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.
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Yoshikazu
Y
Kishino
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan; Japan Society for the Promotion of Science, Chiyodaku, Tokyo, Japan.
Tomohisa
T
Seki
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Jun
J
Fujita
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Shinsuke
S
Yuasa
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan; Japan Society for the Promotion of Science, Chiyodaku, Tokyo, Japan.
Shugo
S
Tohyama
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Akira
A
Kunitomi
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Ryota
R
Tabei
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Kazuaki
K
Nakajima
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Marina
M
Okada
Department of Cardiovascular surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Akinori
A
Hirano
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Hideaki
H
Kanazawa
Department of Cardiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Keiichi
K
Fukuda
D018929:Cell Culture Techniques / Q000379:methods*; D054337:Cell Dedifferentiation / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D007621:Karyotyping; D060888:Real-Time Polymerase Chain Reaction; D029082:Sendai virus; D013053:Spectrophotometry; D013601:T-Lymphocytes / Q000166:cytology* / Q000502:physiology; D019076:Transgenes / Q000235:genetics