A display substrate and a manufacturing method thereof, and a display device relate to the technical field of displaying. The display substrate includes a base substrate; a pixel defining layer; and a common transport layer arranged at one side of the pixel defining layer away from the base substrate, including a first blocking pattern and first transport patterns surrounding the first blocking pattern. An orthographic projection of the first blocking pattern on the base substrate does not overlap with an orthographic projection of the plurality of opening areas on the base substrate. An area of the orthographic projection of the first transport pattern on the base substrate is larger than an area of the orthographic projection of the first blocking pattern on the base substrate.