Mechanisms involved in the developmental organization of the rat striatum were investigated in vitro. The neurons of the patch and matrix compartments were preferentially labeled in vivo with a [3H]thymidine injection on embryonic day (E) 13 or 18, respectively. Two or 7 days later the striatum was removed, dissociated into a single cell suspension and plated on a collagen-coated substrate. After 5 days in culture the neurons had migrated into aggregates. Within an individual aggregate, neurons labeled on E13 tended to clump together, whereas neurons labeled on E18 were randomly dispersed. Comparing between aggregates, [3H]thymidine-labeled E13 cells were located in aggregates containing numerous other labeled E13 cells, whereas [3H]thymidine-labeled E18 cells were dispersed randomly between aggregates. These results suggest that early born striatal neurons (primarily patch cells) selectively associate with each other, and that this process may be crucial to the developmental compartmentalization of the rat striatum.