Definitive hematopoietic stem cells (HSCs) lie at the foundation of the adult hematopoietic system and provide an organism throughout its life with all blood cell types. Several tissues demonstrate hematopoietic activity at early stages of embryonic development, but which tissue is the primary source of these important cells and what are the early embryonic ancestors of definitive HSCs? Here, we review recent advances in the field of HSC research that have shed light on such questions, while setting them into a historical context, and discuss key issues currently circulating in this field.

Elucidating the mechanisms underlying hematopoietic stem cell (HSC) specification and expansion in the embryo has been hampered by the lack of analytical cell culture systems that recapitulate in vivo development. Here, we describe an ex vivo model that facilitates a rapid and robust emergence of multipotent long-term repopulating HSCs in the embryonic AGM region. Because this method includes a cell dissociation step prior to reconstruction of a three-dimensional functional tissue and preserves both stromal and hematopoietic elements, it allowed us to identify the direct ancestry of the rapidly expanding HSC pool. We demonstrate that extensive generation of definitive HSCs in the AGM occurs predominantly through the acquisition of stem characteristics by the VE-cadherin+CD45+ population.

A CD45-negative population of pre-HSCs develops into definitive HSCs in the AGM region of the embryo.

The first definitive/adult-type hematopoietic stem cells (HSCs) in the mouse aorta-gonad-mesonephros region emerge between embryonic days 10.5 and 11.5. The discovery of clusters of hematopoietic cells on the ventral luminal surface of the dorsal aorta in various vertebrate species has led to speculation that the floor of the dorsal aorta may play an essential role for the development of the definitive hematopoietic system. Here, we functionally show affiliation of definitive HSCs with the ventral floor of the dorsal aorta, whereas colony-forming hematopoietic activity is associated with both ventral and dorsal domains. We show that a rare population of PECAM1 high CD45 ؉ cells, within which definitive HSCs reside, is predominantly localized to intraaortic clusters. Furthermore, using ex vivo culture analysis, we demonstrate that the ventral domain of the dorsal aorta has an exclusive functional capacity of inducing and expanding definitive HSCs.aorta-gonad-mesonephros region (AGM region) ͉ embryo