Cell migration plays a crucial role in a variety of physiological and pathological processes. In this study a method of capillary force lithography was used to treat poly(sodium 4-styrenesulfonate) (PSS)/poly(diallyldimethylammonium) chloride (PDADMAC) multilayers with a PDMS stamp before or after etching by NaCl solution, yielding physical patterns with various features such as double thin lines, double strips, meniscus-shaped ridges, and high ridges. The ridge height is controllable in the range of 25 and 1100 nm. Migration of smooth muscle cells (SMCs) was restrained by the double-line patterns in a ridge height-dependent manner. By contrast, the mobility of SMCs was controlled by both the hydration ratio of the multilayers and the pattern features.