Cell adhesive biomaterials have been used for various cells in culture, especially for primary cultures of neurons. Here we examined laminin-111 and its active peptides conjugated to chitosan matrices (ChtMs) for primary culture of rat cortical neurons. Laminin-111 on poly-D-lysine substrate promoted neuronal cell attachment and differentiation. The biological activity of six active laminin-111-derived peptides was examined using a peptide-ChtM construct. When the syndecan-binding peptides, AG73 (RKRLQVQLSIRT, mouse laminin α1 chain 2719-2730) and C16 (KAFDITYVRLKF, laminin γ1 chain 139–150), were conjugated to chitosan, AG73-ChtM and C16-ChtM showed potent neuronal cell attachment activity and promoted axon extension by primary cultured rat cortical neurons. However, the remaining peptides, including integrin-binding peptides, did not show activity when conjugated to ChtM. AG73-ChtM and C16-ChtM also supported neuron survival for at least 4 weeks in serum-free medium without a glia feeder layer. These data suggest that AG73-ChtM and C16-ChtM are useful for primary cultures of central nervous system neurons and have a potential for use as functional biomaterials for tissue engineering in the central nervous system.