Traditional methods to study cell-cell adhesion have been limited by their inability to manipulate cell-cell interactions without simultaneously affecting other microenvironmental factors. Here we describe a novel method that enables the culture of cells with precise simultaneous control of both cell-cell and cell-substratum adhesion. Using microfabricated stamps of poly(dimethylsiloxane), we construct bowtie-shaped agarose microwells into which cells can be cultured. The degree to which cells spread is controlled by the size of the microwell; cell-cell contacts form between neighboring cells within the microwell. This chapter describes the details of stamp fabrication, agarose microwell construction, and cell culture in micropatterned substrata.