We have proposed previously that an increased concentration of growth factor secreted by neurons themselves has a direct effect on survival of the neurons, and thereby cell density serves as a regulator of survival of neurons. In this study, the same idea was used to analyze the aggregation of cerebellar granule neurons in culture experimentally and theoretically. Assuming the transport of growth factor and substrate within an aggregate is by molecular diffusion, the metabolic efficiency of neurons, on the basis of an autocatalytic phenomenon, was increased within an aggregate compared to an identical quantity of dispersed cells. A good agreement in the size of neuronal aggregate between the theoretical prediction and the experimental result was found. This illustrates that growth factors produced by neurons acting in either an autocrine or paracrine manner play an important role during the development of cultured neurons. (C) 2004 Elsevier Ltd. All rights reserved.