A display substrate and display device. The display substrate includes: a base substrate; a plurality of sub-pixel groups, arranged on the base substrate along a row direction and a column direction; and first spacers, second spacers, and third spacers, each of the plurality of sub-pixel groups includes a first sub-pixel, a second sub-pixel, and a third sub-pixel, in one row of sub-pixel groups, each of the first spacers is located between the first sub-pixel and the second sub-pixel which are adjacent to each other, each of the second spacers is located between the second sub-pixel and the third sub-pixel which are adjacent to each other, the third spacer is located between the third sub-pixel and the first sub-pixel which are adjacent to each other, and a number of the first spacers, a number of the second spacers, and a number of the third spacers are approximately equal.