The present disclosure provides a display substrate, including: a base substrate; and a plurality of spacers on a side of the base substrate and each comprising at least one first sub-pattern and at least one second sub-pattern stacked along a normal direction of the base substrate, the first sub-pattern being made of an organic material and the second sub-pattern being made of an inorganic material. The present disclosure also provides a method for manufacturing the display substrate and a display device.