A light-emitting diode (LED) display substrate, a method for manufacturing the same, and a display device are provided and involve the display field. The method for manufacturing the LED display substrate comprises: forming a transparent conductive anode (201) on a substrate (200); forming a pixel region defined by a first PDL (202) and a second PDL (203) on the substrate (200) on which the anode (201) is formed, in which the second PDL (203) made of a hydrophobic material is disposed on the first PDL (201) made of a hydrophilic material; filling a luminescent material into the pixel region to form an emission layer (204) with the luminescent material; and forming a conductive cathode (205) on the substrate (200) on which the emission layer (204) is formed. The manufacturing method allows the luminescent materials to be flatly laid on the LED display substrate so as to improve the luminous quality of the LED display substrate.