The avian thymus is colonized by three waves of hemopoietic progenitors during embryogenesis. An in vivo thymus reconstitution assay based on intrathymic injection of irradiated chicks showed that cells of para-aortic foci were able to differentiate into T lymphocytes, confirming their putative role in the first wave of thymus colonization. This assay was also used to detect and to characterize T cell progenitors from the bone marrow which are involved in the second and third wave of thymus colonization. In the bone marrow, progenitors that differentiated into T cells were found in a subpopulation that expressed the molecules HEMCAM, c-kit and c128. Engraftment of thymus lobes into thymectomized young chick recipients showed that T cell progenitors are replaced in the thymus by subsequent waves of progenitors after hatching. Finally, analysis of thymocyte differentiation suggested that gamma delta and alpha beta T cells migrate from the thymus to the periphery in alternating waves.