The intermediate and anterior lobes of the pituitary gland are derived from an invagination of oral ectoderm that forms Rathke's pouch. During gestation proliferating cells are enriched around the pouch lumen, and they appear to delaminate as they exit the cell cycle and differentiate. During late mouse gestation and the postnatal period, anterior lobe progenitors re-enter the cell cycle and expand the populations of specialized, hormone-producing cells. At birth, all cell types are present, and their localization appears stratified based on cell type. We conducted a birth dating study of Rathke's pouch derivatives to determine whether the location of specialized cells at birth is correlated with the timing of cell cycle exit. We find that all of the anterior lobe cell types initiate differentiation concurrently with a peak between e11.5 and e13.5. Differentiation of intermediate lobe melanotropes is delayed relative to anterior lobe cell types. We discovered that specialized cell types are not grouped together based on birth date and are dispersed throughout the anterior lobe. Thus, the apparent stratification of specialized cells at birth is not correlated with cell cycle exit. Thus, the currently popular model of cell specification, dependent upon timing of extrinsic, directional gradients of signaling molecules, needs revision. We propose that signals intrinsic to Rathke's pouch are necessary for cell specification between e11.5 and e13.5 and that cell–cell communication likely plays an important role in regulating this process. ► The majority of the anterior lobe cell types in the mouse pituitary gland begin differentiation between e11.5 and e13.5. ► The majority of intermediate lobe melanotropes begin differentiation after the anterior lobe cell types between e12.5 and e15.5. ► Regional enrichment of specialized, anterior lobe cell types at birth is not correlated with the timing of cell cycle exit, and potentially involves formation of homotypic networks. ► Intrinsic signaling factors within Rathke's pouch are likely to drive anterior lobe cell specification instead of proposed extrinsic gradients of BMP and FGF signaling.