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. 2022 Jan 27;7:11. doi: 10.1038/s41536-022-00210-1

Fig. 4. SCF and IL-7 promote T-cell proliferation while TNFα inhibits maturation.

Fig. 4

a The RSM was constructed as a six-factor central composite design. Experiments were performed in 7-day intervals until day 42 to measure cytokine responses during all stages of T-cell development. b Cytokine dose responses for proT, CD4ISP, and early DP (CD3-) between days 0–7 and 7–14. Cells were unresponsive to IL-7 until day 7–14 but respond strongly to SCF, IL-3, and TNFα from day 0–7. c Cytokine dose responses for CD4ISP, early DP, and late DP (CD3+), and CD8SP for each 7-day interval between days 14–42. The positive dose-dependent effect of IL-3 and TNFα early in cultures flattens and TNFα begins to inhibit the generation of DP and CD8SP cells. In (b, c), cytokine concentrations were swept from low to high while holding all other cytokines at their scaled center value (0). Shown are square-root transformed cell counts for each population. Experiments used n = 3 pooled UCB donors. d Objectives for optimization of RSM. Populations were either maximized or not included/present in certain 7-day intervals. e Optimized cytokines per 7-day interval (left) or as a three-stage assay (right). Solid lines are the mean of the top five optimizations while dotted lines represent the standard deviation.