Disclosed are a display substrate and a manufacturing method thereof, as well as a display device. The display substrate includes a base substrate, a wire grating polaroid and a light shield layer. The light shield layer is disposed on the base substrate and partly covers a surface of the base substrate. The wire grating polaroid is disposed on the light shield layer at a position corresponding to the light shield layer. The display substrate addresses the problem of reflecting external light by the wire grating polaroid, enhances definition of images viewed by a user and improves user's use experience.