Figure 3

Identification of different cell types in rabbit duodenal tissue and organoids. (a) Rabbit duodenal tissue sections and (b) rabbit duodenal organoids were immunostained for e-cadherin (red), PAS (magenta), Muc5ac (green), soybean agglutinin (SBA) (brown), sucrase-isomaltase (SI) (green) and chromogranin A (ChgA) (green); nuclei were counterstained with DAPI (blue). Boxed regions are shown at higher magnification in the panels below. Pol. epith., polarised epithelial cells. V, villus. GC, goblet cell. PC, Paneth cell. Scale bars = 100 μm (upper panel) and 200 μm (lower panel). (c) Expression of intestinal epithelial maturation-associated genes (MUC5ac, SI, CHGA and LYZ) in differentiated organoids. Data are presented as fold change (2^−ΔΔCt) in gene expression from undifferentiated spheroids, calculated from three individual cell culture wells with three technical RT-qPCR replicates each. Error bars represent standard errors of the mean. Student’s t-test was performed to assess the statistical significance; only statistically significant differences are shown (*p < 0.05). All experiments were conducted in duodenal organoids from a single laboratory rabbit.