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We found the expression of neurofilament was very low in undifferentiated human adult bone marrow mesenchymal stem cells (hMSCs), but its expression could be significantly induced after treatment with combination of growth factors. Approximately 16% of hMSCs differentiated into cells expressing neurofilament after treatment with a combination of FGF and RA, retinoic acid. We also examined the effect of five different cell culture substrates on the expression of neurofilament. One specific combination that was particularly effective in provoking pre-neuronal differentiation was culturing hMSCs on fibronectin-coated dishes and stimulating them with FGF and RA; 40% of cells expressed neurofilament. These results suggest that growth factors and substrates, in combination, can effectively initiate differentiation of hMSCs along a neuronal pathway.