During embryogenesis there is a sequential, temporal appearance of increasingly more-complex hematopoietic cells beginning with unipotential progenitors, proceeding to multipotential (myeloid, erythroid and lymphoid) progenitors and culminating with adult-repopulating hematopoietic stem cells. Current research has established an important role for the aorta-gonads-mesonephros region of the mouse embryo in the generation of multipotential progenitors and hematopoietic stem cells. Comparisons of normal and hematopoietic-cell-mutant mouse embryos have revealed several genes pivotal in hematopoietic stem cell generation/function. Other genes have been implicated in the critical generation of lymphoid lineage potential. Thus, an understanding of the cellular and molecular interactions within the midgestation aorta-gonads-mesonephros region offers insight into the mechanisms of hematopoietic lineage specification during ontogeny and perhaps will lead to a more complete knowledge of the adult hematopoietic system.