A display substrate, a manufacturing method and a display device are provided. The display substrate includes a scan driving circuit; the scan driving circuit includes a plurality of shift register units, at least one shift register unit includes a signal output line and an output circuit, the output circuit includes an output transistor and an output reset transistor; the signal output line includes a first output line portion extending along the first direction; the first output line portion is coupled to the second electrode of the output transistor or the output reset transistor through a plurality of first or second signal line via holes arranged in a signal line overlap area, and the plurality of first or second signal line via holes are arranged along the first direction.