A photosensitive resin composition for cell culture substrates that enables the low-cost manufacture of a cell culture substrate, that can easily form patterns of various shapes when providing a pattern on the surface of a cell culture substrate, has low cytotoxicity, and that can form a cell culture substrate; a cell culture substrate that is formed using the photosensitive resin composition; and a cell culture substrate manufacturing method that uses the photosensitive resin composition. The photosensitive resin composition includes a photopolymerizable monomer and a photopolymerization initiator. The photopolymerizable monomer contains a defined amount of a polyfunctional monomer that is at least trifunctional, and the content of the photopolymerization initiator is within a prescribed range.