Extended Data Fig. 2: Piglet scaffold characterization and extended human scaffold characterization.
a, Representative H&E images and DAPI immunostainings of a native piglet intestine and following one and two cycles of DET. Scale bars represent 100µm. b, Quantification of DNA, glycosaminoglycans (GAGs) and collagen per milligram of wet tissue in native piglet intestine and following one and two cycles of DET. Data represent mean ± s.e.m of n = 3 biologically distinct piglet intestine samples (DNA and GAG quantification) and of n = 2 biologically distinct samples (collagen quantification). One-way ANOVA with Dunnett’s multiple comparisons test: DNA, ****p <0.0001 (Native versus cycles I and Native versus cycle II); GAG, p=0.4055 (Native and cycle I) and **p=0.0036 (Native and cycle II). The experiment was performed once. c, Representative histology images of Elastic Van Gieson (EVG) and Alcian blue (AB) stainings confirm preservation of elastin and GAGs respectively following one and two cycles of DET. Masson’s trichrome (MT) and Picro-sirius red (PS) histological stainings confirms maintenance of connective tissue and collagens following one and two cycles of DET. Scale bars represent 200µm. d, Representative immunohistochemical staining for Collagen I (Col-1), Collagen IV (Col-4), Fibronectin, Laminin indicating the preservation of these ECM proteins in a piglet scaffold following two cycles of DET. Scale bars represent 100µm. Images are representative of 3 biological replicates. e, Stress-strain curves of human colon [patients 19, 20, 21] and small intestinal [patients 21, 23, 24] scaffolds and calculated Young’s modulus. Data represent mean ± s.e.m. of n = 3 biologically distinct patient samples. No significant difference identified between human SI and colon samples; unpaired t test, p=0.5871. The experiment was performed once. f, PC1 loading plot associated with the PC1 vs PC2 scores plot shown in Fig. 2f. The spectral features show distinct biochemical differences that facilitate the differentiation of the mucosal region of the SI [patient 1] and colon [patient 18] from the remaining intestinal layers (submucosa and muscularis propria). The intensity of the corresponding peaks found within the loadings plot indicate their influence on the separation in the scores plotted along the associated axis. g,h, Representative immunohistochemical staining of the native pediatric SI [patient 1] and colon [patient 18] tissue using the indicated antibodies, n = 2 biologically distinct patient tissue samples. Scale bars represent 100μm.