The present disclosure provides a display substrate, a manufacturing method thereof and a display device having display units on connected islands. The display substrate includes a base substrate, including a hollowed-out region, an island and a connection bridge; a plurality of display units; and a source/drain metal pattern, including patterns of a plurality of signal lines. On the island, a plurality of inorganic film layers is arranged between the base substrate and the source/drain metal pattern, and the plurality of inorganic film layers includes a first inorganic film layer arranged at a side furthest away from the base substrate. On the connection bridge, at least one first organic film layer is arranged between the base substrate and the source/drain metal pattern.