The present disclosure provides a display substrate, a manufacturing method thereof, and a display device. The display substrate includes a base substrate and a plurality of subpixels arranged on the base substrate in an array form. The plurality of subpixels includes a plurality of display subpixels at a display region of the display substrate and a plurality of virtual subpixels, and at least a part of the virtual subpixels are arranged adjacent to the display subpixels. The virtual subpixel includes a first potential signal line pattern, a virtual subpixel driving circuit including a virtual driving transistor and a first conductive connection member coupled to a gate electrode of the virtual driving transistor, and a second conductive connection member coupled to the first conductive connection member and the first potential signal line pattern.