The present disclosure provides a display substrate and a manufacturing method thereof, and a display device. The display substrate includes a plurality of pixel regions, each of the pixel regions including a display region provided with a pixel electrode and a driving region provided with a pixel circuit. The pixel circuit includes at least one pixel transistor having a first electrode and a second electrode which are coupled to an active layer of the at least one pixel transistor through connection vias, respectively. The driving region is further provided with a first plate of a storage capacitor, the first plate is insulated from and overlapped with the first electrode and the second electrode of the pixel transistor in a direction perpendicular to the display substrate, and the first plate is provided with openings at positions corresponding to at least some of the connection vias.