Extended Data Fig. 3: Monitoring CAR T cell dynamics on-chip.
(a-c) Representative images showing CAR T cell extravasation (a), infiltration (b), and the process of killing a Reh B-ALL cell by a CAR T cell. CAR T cell in red, Reh B-ALL in green, and vessel (HUVECs) in blue. Representative images were from one of three independent experiments with similar results (n = 3). (d,e) ELISA measured CAR T cell secretions of granzyme B (d) and perforin (e) on day 2 from leukaemia chips established with CD19-expreesing leukaemia (CAR + CD19+), non-CD19 expressing leukaemia on-chip (CAR + CD19-), or no leukaemia (CAR+None) or from CAR T cell cultured off-chip (CAR off-chip). Data was collected from eight independent experiments (n = 8). Two-way ANOVA followed by Tukey’s post hoc test, mean and s.e.m. (f,g) Flow cytometry measurement of surface expressions of T cell activation markers CD25 (f) and CD69 (g) on CAR T cell and mock T cell after on-chip infusion for 2 days. Data was collected from five independent experiments (n = 5). Unpaired, two-sided, Student’s t-test, mean and s.e.m. (h) Gating strategy for identification of live CD3 + T cell using CAR T cell sample as an example. (i-k) ELISA measured secretions of INF-γ (i), granzyme B (j), and perforin (k) from leukaemia chips respectively treated with CAR T cell (CAR) and Mock T cell (Mock) at different time points (12 h, 24 h, and 48 h). Data was collected from four independent experiments (n = 4). Unpaired, two-sided, Mann-Whitney test, mean and s.e.m.
