A light-emitting substrate, a method of manufacturing a light-emitting substrate, and a display device are provided. The light-emitting substrate includes: a first substrate, wherein the first substrate includes a first base substrate, a light-emitting diode arranged on the first base substrate, and a first conductive pad arranged on the first base substrate; a second substrate arranged opposite to the first substrate, wherein the second substrate includes a second base substrate, and a second conductive pad arranged on the second base substrate; and a bonding wire structure including a bonding wire, wherein the first conductive pad is located on a surface of the first substrate away from the second substrate, the second conductive pad is located on a surface of the second substrate away from the first substrate, and the bonding wire is configured to electrically connect the first conductive pad and the second conductive pad.