In vitro meat is an edible muscle tissue produced in a laboratory using bioengineering tools, avoiding the slaughter of farmed animals. In the early stages of in vitro meat production, the muscle cells need to be oriented and aligned to form muscle fibers. Additionally, the adherent muscle cells require an appropriate matrix where they can proliferate and differentiate. In this work, a film (matrix) was formulated with edible biopolymers and then shaped using micropattern mold and cold-casting technique. The mold was fabricated by using a laser cutter, allowing building parallel microchannels of ~ 70 μm wide. The film surface was suitable for laying muscle cells in a fiber-like array. Microstructural characterization of the micropatterned structures was done by using stereoscopic microscope and scanning electron microscopy. The micropatterned films displayed high biocompatibility, allowing muscle cell adhesion (~ 70%) and growth (doubling time ~ 18 h). Fluorescence microscopy was used to study the morphology of the cells cultured onto the micropatterned films, showing fiber-like morphology (~ 80-μm fiber diameters). The shape and size of the obtained cells mimic the muscle fiber. By using immunofluorescence microscopy, the expression of specific myogenic markers was identified. These results confirm that edible films made by casting on micropatterned molds can be used for in vitro meat production.