A display substrate is provided, including: a base (1); and a plurality of pixel units (10) located on the base (1) and arranged in an array, where the display substrate further includes a plurality of data lines (25), and for each of the plurality of data lines (25), the data line (25) extends along a column direction and is located between adjacent first pixel unit (10A) and second pixel unit (10B) in a row direction, the data line (25) is arranged in a different layer from a first pixel electrode (40A) of the first pixel unit (10A) and a second pixel electrode (40B) of the second pixel unit (10B), and the data line (25) includes a first branch line (252) and a second branch line (253) that are connected in parallel and extend in the column direction; an orthographic projection of the first branch line (252) on the base (1) at least partially overlaps with an orthographic projection of the first pixel electrode (40A) on the base (1); an orthographic projection of the second branch line (253) on the base (1) at least partially overlaps with an orthographic projection of the second pixel electrode (40B) on the base (1). A display panel and a method of manufacturing a display substrate are further provided.