Three-dimensional imaging of whole midgestation murine embryos shows an intravascular localization for all hematopoietic clusters.

Hematopoietic cell clusters associated with the midgestation mouse aorta, umbilical and vitelline arteries play a pivotal role in the formation of the adult blood system. Both genetic and live-imaging data indicate that definitive hematopoietic progenitor/stem cells (visualized as clusters) are generated from hemogenic endothelium. A 3-dimensional (3-D) whole embryo immunostaining and imaging technique has allowed quantitation and cartographic mapping of intravascular hematopoietic clusters. During this period the vitelline artery is most extensively remodeled, and several reports have suggested that vitelline remodeling leads to extravascular hematopoietic cluster emergence. Whether the earliest definitive progenitors/stem cells are intra or extra vascular could influence the process by which these cells migrate to the next hematopoietic territory, the fetal liver. Hence, by 3-D imaging we more closely examined hematopoietic clusters in the vitelline and associated connected small vessels and show here that hematopoietic clusters (particularly large clusters) are intravascular during the period of vascular remodeling.