In order to establish an in vitro attachment culture method for obtaining highly purified neural progenitor cells (HPCs) from the hippocampus of adult Wistar rats and to identify functional L-type calcium channels in those cells, the adult rat hippocampal tissue was dispersed into a single cell suspension, and the dissociated cells were cultured in serum-free DMEM/F12 medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), N2 and B27 supplement. After six passages, the proportion of nestin-positive cells reached to 99.9%. Following fourteen-days culture in differentiation medium, neuron-like and astrocyte-like cells were observed, which expressed β-tubulin Ⅲ (Tuj1) and glial fibrillary acidic protein (GFAP), respectively. Immunofluorescent double labeling and Western blot showed an expression of Cav1.2α1C and Cav1.3α1D subunits in HPCs, and functional L-type calcium channels were confirmed by confocal microscopic Ca2+ imaging. Moreover, L-type calcium channel currents were recorded in those cells by using whole-cell patch clamp techniques. The results indicated that adult rat HPCs express functional L-type calcium channels.