Figure 3.

Pericytes from hPSC, brain, and placenta promote the formation of CD4⁺CD25^highFoxP3⁺CD127⁻ T cells. (A): Representative dot plots of CD25 and FoxP3 expression of gated CD4 T cells from cocultures of peripheral blood mononuclear cells or peripheral blood (PB) CD3⁺ T cells with hPSC, brain, and placenta pericytes after 3 days of induction. (B): CD3⁺ T cells were separated into CD3⁺CD25⁺ and CD3⁺CD25⁻ subsets, and then the CD3⁺CD25⁻ subset was used for pericyte induction experiments. The expression of CD4 and CD25 and the formation of CD4⁺CD25^high populations among the allostimulated CD25⁻ T-cell subset are shown in representative dot plots. (C): Percentages of CD4⁺CD25⁺ and CD4⁺CD25^high cell populations generated from the CD3⁺CD25⁻ T-cell subset in hPSC and brain pericyte cocultures. Mean ± SEM of triplicate samples is shown from seven independent experiments with seven different PB CD3⁺CD25⁻ T cells and at least four pericyte cell lines per experiment. ∗, p < .05 compared with pericyte-stimulated T cells of a matched subset. (D): Quantitative polymerase chain reaction gene expression of FoxP3 in the CD3⁺CD25⁻ T-cell subset compared with the CD3⁺CD25⁻ T-cell subset after 3 days of cultivation with hPSC and brain pericytes. Data are mean ± SEM of 3 independent experiments with PB cells from 2 different donors. ∗, p = .05 compared with pericyte-stimulated CD25⁻ T cells. (E): Representative expression of FoxP3 in gated CD4⁺CD25⁺ population, which was induced by pericyte from the CD3⁺CD25⁻ subset within 72 hours. CD127 expression was evaluated on CD4⁺CD25⁺FoxP3⁺ and CD4⁺CD25⁺FoxP3⁻ subpopulations. Abbreviation: MNC, mononuclear cell.