Early haemopoietic stem cells in the avian embryo

Using 'yolk sac chimaeras', we have previously demonstrated that stem cells, destined to colonize haemopoietic organs other than the yolk sac, arise in the embryo proper. We have now investigated the emergence and potentialities of these cells in vivo and in vitro. The in vivo approach consisted of interspecies grafting between quail and chick embryos. The cell progeny from the grafts was detected by means of QH1, a monoclonal antibody specific for the quail haemangioblastic lineage. When grafted into the dorsal mesentery of the chick embryo, which is a haemopoietic microenvironment, the region of the aorta from E3-E4 quail embryos generated large haemopoietic foci. When associated with a chick attractive thymic rudiment, cells left the quail aorta, entered this rudiment and underwent lymphopoiesis. Cell suspensions prepared from 40-50 chick aortae, seeded in appropriate semi-solid media, yielded macrophage, granulocyte or erythrocyte clones. These colony forming cells were two to eight times more frequent than in cell preparations from hatchling bone marrow. By contrast, cells prepared from the whole embryonic body deprived of the aorta were not clonogenic. By interspecies grafting of somatopleural (ectoderm + mesoderm, e.g. limb bud) or splanchnopleural rudiments (endoderm + mesoderm, e.g. lung, pancreas, intestine), the endothelial lining of blood vessels was shown to arise by two entirely different processes according to the rudiment considered: angiogenesis, i.e. invasion by extrinsic endothelial cells, in the limb bud, and vasculogenesis, i.e. in situ emergence of endothelial cells, in internal organs. The spleen, which first develops as a continuum to the pancreatic mesoderm, acquires its endothelial network by vasculogenesis, and is colonized by extrinsic haemopoietic stem cells. Granulopoietic cells in the pancreas and accessory cells in the lung are also extrinsic. Thus, in the case of endomesodermal rudiments, interspecies grafting reveals separate origins of endothelial and haemopoietic cells.