A method of manufacturing a patterned substrate for culturing cells. The method includes the steps of: (1) preparing a substrate, (2) forming a first plasma polymer layer by integrating a first precursor material on the substrate using a plasma, wherein the first plasma layer inhibits cell adsorption, and wherein the first precursor material is a siloxane-based compound having a siloxane functional group with the Si—O—Si linkage, (3) placing a shadow mask having a predetermined pattern on the first plasma polymer layer thus formed, and (4) forming a second patterned plasma polymer layer by integrating a second precursor material using a plasma, wherein the second patterned plasma layer permits culturing of cells, whereby the patterned substrate is obtained.