Provided are a display substrate, a display device and a method for manufacturing a display substrate. The display substrate (10) comprises a base substrate (4) and a plurality of spacers arranged on the base substrate (4), wherein an end, away from the base substrate (4), of each of the plurality of the spacers is provided with an inclined surface; and the directions of inclination of the inclined surfaces of the plurality of the spacers are consistent. When rubbing alignment is performed on an alignment layer on the display substrate, poor display caused by relatively significant deformation of fibers on a surface of a rubbing cloth when passing through the spacers can be avoided.