The present invention provides a tissue construct-forming substrate for forming a three-dimensional tissue construct containing proliferating cells, the substrate including a porous film having through-holes, and the porous film having, on the surface of the film, a cell adhesive region capable of retaining cells and a cell non-adhesive region located at a peripheral region of the cell adhesive region, a tissue construct-forming kit comprising the above-mentioned tissue construct-forming substrate and a frame, and a method for forming the above-mentioned tissue construct.