The control of neuronal cell adhesion and growth in artificially defined networks in vitro was developed for the study of neuronal signals propagation. Microcontact printing (μCP) with PDMS microstamps was used as the chemical method to control cell adhesion and growth into defined networks. For controlled hippocampal neuronal cell growth, cytophilic poly-d-lysine (PDL) was patterned on glass substrates by μCP and then cytophobic PEG was self-assembled for passivation. The patterns were identified by fluorescent microscopy, atomic force microscopy, and condensation figure. The positively charged surface of PDL patterns interacted electrostatically with the negatively charged moieties of the cell membrane surfaces so that the attachment of the cells to the substrate surface was enhanced. The cultured hippocampus cells grew quite selectively along the printed PDL tracks. The track width suitable for forming single neuronal growth was discovered to be less than about 10 μm.