Fig. 6 H&E staining of in vitro intestine models grown in static and perfusion culture for 7 and 14 days. HDFn

fibroblasts were cultured in Alvetex Scaffold to create a lamina propria-like compartment. Caco-2 cells were

then cultured on the Scaffold for 21 days until a mature epithelial layer formed. These mature intestinal

models were cultured for a further 7–14 days in static and perfused conditions. Perfused intestine models

exhibit crypt–villus morphogenesis. Caco-2 cells form a polarized layer on the surface of the villus structures.

Larger villus-like structures form following 14 days of perfusion compared to 7 days. Scale bars: 100 μm

Fig. 7 Immunofluorescence staining of static and perfused 3D intestine models. Static intestine models show

no Ki67 staining in the epithelial layer. Perfusion of intestine models results in Ki67 positive staining restricted

to the basal layer of the crypt structures, suggesting proliferation is occurring. E-cadherin staining is evident

throughout the epithelial layer in both static and perfused models, indicating mature junctional complexes

form between epithelial cells. Alpha smooth muscle actin staining in fibroblasts increases within the lamina-

propria-like compartment as a result of perfusion. Scale bars: 50 μm