Display omitted •A Janus film with a ridge-like rough surface and a flat surface was fabricated via cryogenic electrospinning.•The rough and flat surface of the Janu film mimic the natural anatomical surface texture of epithelium and lamina propria, respectively.•The Janus film allows the paracrine action between epithelial cells and fibroblasts while keeping the insulation between them. Substantial mucosa defects in gastrointestinal tracts are difficult to repair, causing high mortality. Existing artificial mucosa studies overlooked the natural surface texture and the paracrine action between epithelial cells and fibroblasts, leading to an incomplete repair of the mucosa. Here, a Janus film with an exterior surface having a ridge-like texture and an interior flat surface to mimic the anatomical pattern of natural mucosa in digestive tracts is obtained through cryogenic electrospinning (e-spinning). Inspired by the natural mucosa surface textures, biopolymer nanofiber assemblies are employed to replicate the micro-ridges of the exterior layer of mucosa, while the flat surface mimics the morphology of the inferior layer. Hence, the exterior surface of Janus film bolsters epithelial cells' proliferation through the macro pores between fiber assemblies and the nanopores on fibers. In addition, the Janus film realizes a bilayer epithelium/lamina propria reconstruction by culturing epithelial cells and fibroblasts on different sides of the Janus film. Moreover, the Janus film assists the paracrine action between epithelial cells and fibroblasts while insulating their direct contact, an essential mimicry of the natural counterpart. In summary, it is proved that the Janus film will be a promising repair solution for mucosa defects in digestive tracts.