Fig. 6: Assembly and testing of a T cell containing BM-MPS for TCB safety assessment.

Mean values +/− SD of two to five circuits (n = 2–5) of one CD34+ cell donor (N = 1) are shown if not specified otherwise. A Number of sampled T cells from the chip at day 21 and day 24 in the presence of absence of IL-2 relative to the amount of seeded naive or preactivated T cells at day 17. B Percentage of naive or preactivated CD4+ and CD8 + T cells that express activation markers CD69, PD-1, CD25, and HLA-DR before chip culture (day 17) and after 7 days cultivation on chip without additional stimulation (d24) in the presence of absence of IL-2. C–E Sampled HSPC (C), erythroid cell counts (D) and granulocytes (E) at day 24 in circuits without T cells and in circuits with naive or preactivated T cells in the presence or absence of IL-2. F Time schedule of 7 day treatment with ESK-1-like TCB and an isotype control TCB at a concentration of 1 µg/mL in the Chip2 system with cell sampling for flow cytometry analysis after four and seven days after treatment start. G Sampled CD3 negative non-T cell counts from circulation and the ceramic scaffold at day 24 in circuits treatedwith the ITC-TCB or ESK1-TCB. Counts are normalized to the mean of untreated control circuits. Circuitsthat were supplemented with IL-2 are marked by squares, circuits without IL-2 supplementation with circles. H Expression of activation markers CD69, PD-1, CD25, and HLA-DR on naive CD4+ and CD8 + T cells at day 21 and day 24 treated with the ITC-TCB or ESK1-TCB.