During early postnatal development in the rat, the tissue architecture of the pituitary gland shows changes, revealing an intense migration process of the cells. The aim of this work was to examine anterior pituitary cell migration over type I and III collagen as well as type IV collagen, of cultured pituitary cells from infantile rats and adult rats, and the participation of the epidermal growth factor in this process. Differences in cell migration rate over these two types of collagen substrates were observed at both ages, and all in all, three times more cells migrated over type I/III collagen than over type IV collagen. These data show the migration-promoting role of type I/III collagen for pituitary cells. Furthermore, when infantile cells were challenged to migrate over bovine serum albumin, the migration rate diminished, and, on the contrary, adult cell migration was higher. However, over collagen, infantile cells increased their migration rate with epidermal growth factor stimulation and adult cells showed a decrease in migration when the growth factor was in the medium. During migration, pituitary cells associated and arranged in clusters. This behavior increased in the presence of epidermal growth factor in the infantile cultures. Moreover, epidermal growth-factor-stimulated infantile cells formed larger aggregates. Adult cells also showed associative behavior, but more cells were observed isolated than in cluster arrangements and the growth factor did not induce changes in this behavior. Results showed a difference in the response of cell migration and cell association capacity to epidermal growth factor after migration of infantile and adult pituitary cells. With these observations we propose that epidermal growth factor is a cell regulator of the pituitary tissue re-arrangement process during the infantile period.