In Vitro Generation of Stem Cell Memory-Like T Cells from Activated T Cells

Makoto Ando, Mari Ikeda, Akihiko Yoshimura, Taisuke Kondo

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)


Adoptive T-cell therapy is an attractive strategy for cancer immunotherapy. The transfer of in vitro expanded tumor-associated antigen (TAA)-specific T cells from patients may effectively fight against the original tumor cells. The chimeric antigen receptor-engineered T (CAR-T) cells are also shown to be a promising therapy for hematologic malignancies. However, one of the limitations of these T-cell-based therapies is a rapid acquisition of tolerant (anergy, deletion, dysfunctional and/or exhausted) phenotypes of T cells during activation in vitro and/or after transfer in vivo. We and others found that stem cell memory T (TSCM) cells are strongly resistant against such tolerance, showing strong expansion and persistence in vivo, and provide long-lasting antitumor effects. Here we describe a protocol for the generation of phenotypically TSCM-like cells (iTSCM cells), which can be induced by simple co-culture of activated T cells with OP9 stroma cells expressing a Notch ligand. We also showed the methods of cancer immunotherapy by using NSG mice.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages13
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Adoptive T-cell therapy
  • Co-culture with feeder cells
  • EB virus-specific T cells
  • NSG mice
  • Notch signaling
  • Stem cell memory T cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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