Poly-D-lysine, poly-L-lysine, collagen, laminin, fibronectin, and Matrigel were compared with standard tissue grade polystyrene for their impact on the expansion and neuronal differentiation of mesenchymal stem cells (MSCs). Among these substrates, adsorption of Matrigel at 5 microg/cm2 did not enhance cell proliferation but gave rise to the highest percentage of MSC-derived neuron-like cells with the best morphological differentiation. Matrigel at a higher coating density of 50 microg/cm2 not only further enhanced the differentiation but also significantly improved cell expansion. In contrast, poly-D-lysine did not effectively support the growth of MSCs. Hence the expansion and neuronal differentiation of MSCs both depend on surface properties of the culture substrate. These results could lead to a culture process with improved yield of MSC-derived neuron-like cells and to novel biomaterials for tissue engineering.