Three-dimensional cell aggregation enhances growth-promoting activity of NGF in adult DRG

After removal of the connective tissues with collagenase-trypsin treatment, DRG explants without nerve fibres from mature rodents regenerated numerous neurites all around the explants in response to NGF. To observe the response in the absence of any non-neuronal cells in the ganglia, we introduced three-dimensional cell aggregates consisting of variable numbers of highly purified isolated adult DRG neurones. In this culture system, the addition of NGF to the medium increased the maximum neurite length, the number of neurites, and the cell survival rate. Furthermore, the magnitude of this effect increased with cell aggregate size. Our results suggest that the addition of NGF produces extensive neurite outgrowth in three-dimensional aggregates of matured sensory neurones in the absence of any non-neuronal cells.