A display substrate, a display device having the same and a method of manufacturing the display substrate are provided. The display substrate includes a base substrate having a pixels-populated area (PA) and a surrounding area (SA) outside the PA, a first contact pad portion formed in the surrounding area, a second contact pad portion formed in the surrounding area formed to be spaced apart from the first contact pad portion with a spacing region provided therebetween, an insulating layer formed in the spacing region between the first and second contact pad portions and having a thickness smaller than or equal to a thickness of each of the first and second contact pad portions, and a first conductive film formed on the first and second pad portions.