A display device, a display panel, and a manufacturing method thereof are proposed. The display panel includes a substrate (1), a first insulation layer (2), a first electrode layer (3), a pixel definition layer (4), a light-emitting function layer (5) and a second electrode (6). The first insulation layer (2) is provided on a side of the substrate (1). The first insulation layer (2) comprises a plurality of pixel regions (202) distributed in an array and a seperation region (201) separating the pixel regions (202). The first electrode layer (3) is arranged on the surface of the first insulation layer (2) away from the substrate (1), and includes a plurality of first electrodes (31). The orthographic projection of each first electrode (31) on the first insulation layer (2) is located within a respective pixel region (202). The pixel definition layer (4) is arranged on the surface of the first insulation layer (2) away from the substrate (1), and exposes each first electrode (31). The pixel definition layer (4) is formed with a pixel definition groove (41) in the area corresponding to the seperation region (201), and the middle part of the pixel definition groove (41) has a first protrusion (42) protruding in a direction away from the substrate (1). A sub-groove (40) is formed between the sidewall of the first protrusion (42) and the sidewall of the pixel definition groove (41). The light-emitting functional layer (5) covers the pixel definition layer (4) and the first electrodes (31) and the first insulation layer (2) exposed by the pixel definition layer (4). The second electrode (6) covers the light-emitting functional layer (5).