Various protocols have been published for in vitro expansion and maintenance of adult neural progenitor cells (ANPC). However, there are only few data comparing these protocols regarding their influence on proliferation, migration and differentiation. Freshly isolated ANPC from olfactory bulb (BO) and dentate gyrus (DG) of adult rat brains forming neurospheres and expressing the neural stem cell markers nestin and Sox-2 were used in a comparative analysis of five different medium combinations. Medium containing N2 and fetal calf serum (FCS), but no additional cytokines was unsuitable for an effective long-term expansion of ANPC due to a significantly reduced proliferation rate. Media containing BIT, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), platelet-derived growth factor AB (PDGF-AB) and leukemia inhibitory factor (LIF) or B27, bFGF and EGF are recommendable for the cultivation of DG-derived ANPC as neurospheres only. Unlike, culture media containing BIT, bFGF, EGF and PDGF-AB or N2, bFGF and EGF were suitable for all applications tested as they responded similarly regarding proliferation, migration and expression of differentiation markers. The results of the present study might help to improve the effective in vitro expansion of ANPC derived from rare human tissue samples.