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Fig. 1 | Journal of Translational Medicine

Fig. 1

From: A short CD3/CD28 costimulation combined with IL-21 enhance the generation of human memory stem T cells for adoptive immunotherapy

Fig. 1

Gating strategy for the identification of human TSCM. T cells were identified by gating on lymphocytes (SSC versus FSC), singlets (FSC-H versus FSC-A) and live T cells (SSC versus LIVE/DEAD). CD4+ and CD8+ T cells were simultaneously identified with anti-CD4 and anti-CD8 antibodies. a Gating strategy of fresh blood cells. After gating on CD4+ and CD8+ cells, TCM and TEM subpopulations were identified based on CCR7 and CD45RO expression. In the gated CCR7+CD45RO population, cells expressing CD45RA and CD27 were further analyzed. In this later population (CCR7+CD45ROCD45RA+CD27+), TN and TSCM were identified based on the CD95 expression. TN is defined as CCR7+CD45ROCD45RA+CD27+CD95 whereas TSCM subpopulation is defined as CCR7+CD45ROCD45RA+CD27+CD95+. Red arrows indicate the sequential gating strategy. b Gating strategy of 10 days culture cells. After gating on CD4+ and CD8+ cells, TCM and TEM subpopulations were identified based on CCR7 and CD45RO expression. In the gated CCR7+CD45RO population, cells expressing CD45RA and CD27 were further analyzed. In this later population (CCR7+CD45ROCD45RA+CD27+), TSCM were identified based on the CD95 expression. TSCM subpopulation is defined as CCR7+CD45ROCD45RA+CD27+CD95+. Similarly, in the gated CCR7+CD45RO+ population, cells expressing CD45RA, CD27 and CD95+ identify a TSCM-like subpopulation, which is defined as CCR7+CD45RO+CD45RA+CD27+CD95+. Red arrows indicate the sequential gating strategy

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