Recently, microfabrication tools have been utilized to quantify the role of the cellular microenvironment on cell activity and function. Improving tissue regeneration by cell culture on scaffold material will also require tools to control cellular organization in 3-dimentional (3-D) condition. Our objective was to improve cartilage tissue engineering using 3-D cell organization technology. In this study, we developed an anisotropic cartilaginous tissue by cell patterning within hydrogel slabs using dielectrophoretic (DEP) forces. Our data indicate that the embedded chondrocytes remained viable and reconstructed cartilaginous tissue along the patterned cell array. DEP cell patterning may become a useful approach for reconstructing anisotropic structure in cartilage regeneration.