This study presented facile method of cell patterning using fabricated PDMS patterns on polyelectrolyte coated surface. This basic principle is the fabrication of functional surface presenting two orthogonal surfaces such as cell adhesive and repellent properties. Cell adhesive surface was firstly fabricated with simple coating of polyelectrolyte multilayer. And then, the desired patterns of PDMS for the prevention of nonspecific binding of cells were transferred onto the previously formed thin film of polyelectrolyte multilayer. Thus, we could prepare novel functional surface simultaneously containing PDMS and polyelectrolyte region. As expected, the PDMS regions showed effective prevention of nonspecific binding of cell and the other region, exposed polyelectrolyte area, provided cell adhesive environment. The height of formed PDMS structure was about 100 nm. Based on this method, cell patterning can be successfully obtained with various pattern shapes and sizes. Therefore, we expect that this simple method will be useful platform technology for the development of cell chip, cell based assay system, and biochip.