Abstract: Objective To study whether the neuron-like cells derived from bone marrow stromal cells (BMSCs) may excrete amino acids with neurobiological activities and possess the biochemical characteristics of neurons. Method Under sterile condition, BMSCs from New Zealand rabbits were purified by gradient density centrifugation, and were induced to differentiate into neural stem cells and neuronal-like cells in the culture medium for neural stem cells containing retinoic acid (RA, 0.5 μg/ml) and glial-derived neurotrophic factor (GDNF, 20 ng/ml). The differentiated cells were then examined with immunocytochemical method and high-performance liquid chromatograpy (HPLC). Results The round and enlarged BMSCs on day 10 of cell culture were positive for nestin, and on day 20, the cells with RA+GDNF stimulation differentiated into neuron-like cells with long protrusions and presented neuron-specific enolase (NSE) antigen. HPLC identified high levels of amino acids like Asp, Glu, Gly and Ala in the differentiated cells (P<0.01). Conclusions Rabbit BMSCs may proliferate in vitro as from nestin-positive cells and differentiate into NSE-positive cells containing high levels of excitatory and inhibitory amino acid neurotransmitters. RA and GDNF are important promoters for in vitro differentiation of the BMSCs toward neural stem cells.