A cell culture substrate including: a substrate; and a layer containing hydrophilic polymer with a layer thickness of 5 to 2,000 nm covering at least a part of a surface of the substrate, in which the hydrophilic polymer contain a phosphorylcholine group or a hydroxyl group, the cell culture substrate has regions (A) below and regions (B) below, and an unevenness height at a boundary between each of the regions (A) and each of the regions (B) is 1 to 500 nm.



(A): Island-shaped region of 0.001 to 5 mm2 in area with cell adhesiveness and cell proliferation properties
(B): Region which is adjacent to region (A) and has no cell adhesiveness or cell proliferation properties