Fig. 2: Intestinal villus atrophy and barrier compromise.
a, DIC imaging of intestine chips, top-down view (scale bar, 50 µm). b, Immunofluorescence cross-section micrographs showing villus-like structures in the intestine chips. Yellow, phalloidin; Blue, Hoechst; Dashed white line, upper surface of chip membrane. Scale bar, 50 µm. c, Villus-like structures height differences between control and −N/−T, healthy (mean: 211 µm for control and 60 µm for −N/−T) and EED (mean: 288 µm for control and 48 µm for −N/−T) intestine chips. Healthy, P < 0.000001, EED, P = 0.000685, multiple Student’s t-test. Each datapoint on the graph represents the average of 4 to 5 measurement points (membrane to top of the villi) measured in 2 to 3 cross-sectional images of the chip in at least 2 chips per condition. Two different healthy donors and one EED donor were used. Each chip corresponds to one biological replicate. d, Immunofluorescence microscopic views of longitudinal section through the two-channel intestine chips showing the mucus layer overlying the epithelium in the upper channel stained with Alexa 488-conjugated lectin. Scale bar, 250 µm. e, Graph showing differences in mucus thickness between control and −N/−T, healthy (mean: 448 µm for control and 243 µm for −N/−T) and EED (mean: 533 µm for control and 221 µm for −N/−T) intestine chips. Each datapoint on the graph represents the average of 2 measurement points measured in views similar to those shown in d; 2 chips per condition with chips created from cells from two different healthy donors and one EED donor. Healthy, P = 0.004092; EED, P < 0.000001, multiple Student’s t-test. Each chip corresponds to one biological replicate. f, Graph showing differences in Papp between control and −N/−T, healthy (mean: 3.82 × 10−7 for control and 3.41 × 10−6 for −N/−T) and EED (mean: 5.71 × 10−7 for control and 1.41 × 10−6 for −N/−T) intestine chips (9 for healthy chips and 8 for EED intestine chips). Healthy, P = 0.000005; EED, P = 0.001382, multiple Student’s t-test. Each chip corresponds to one biological replicate. Graphed data in c, e and f are mean ± s.e.m.
