Fig. 5.

(A) Three-dimensional confocal reconstruction of coculture of Ishikawa cells and tHESCs stained for live and dead cells. The transition of flat to folded states after relaxation of the prestretched substrate with ${\lambda }_{ps}=2$ is demonstrated. Note that due to attenuation of fluorescent signal in the hydrogel, two sets of z-stack images are taken at different gains to capture epithelial and stromal components separately before they are integrated into one single 3D reconstruction image. (B) Three-dimensional confocal reconstruction of cocultured artificial mucosa which is fixed and stained. Encapsulated tHESCs are stained with phalloidin for F-actin in GelMA while Ishikawa cells are stained with both CK18 and phalloidin on the surface of GelMA. Nuclei are stained with Hoechst. Note that the staining of phallodin is overexposed to better visualize the encapsulated tHESCs due to signal attenuation in the presence of epithelium.