The hematopoietic developmental hierarchy originates with a rare population of lymphohematopoietic stem cells that are capable of extensive self-renewal as well as the continuous generation of more developmentally restricted progeny. The generation of mature blood cells from these pluripotent hematopoietic stem cells involves the highly regulated progression through successive stages involving commitment to a specific cell lineage, terminal differentiation of lineage-restricted progenitors, and growth arrest. Although the differentiation commitment of stem cells may be intrinsically determined, it is apparent that a wide variety of external and internal stimuli can influence and modulate lineage choice and differentiation. These factors cooperate with cellular transcription factors to activate or repress the expression of genes responsible for lineage choice, diverse mature phenotypes, and cell cycle progression. The extrinsic and genetic mechanisms that orchestrate the differentiation commitment and myeloid lineage restriction of pluripotent stem cells are of fundamental importance in the regulation of hematopoiesis. The elucidation of these mechanisms of normal myeloid differentiation has provided instrumental insights into the biology of leukemia and other hematopoietic disorders.