Abstract
The midgestation mouse head harbors hematopoietic stem cells (HSCs) at the same time HSCs are detected in the aorta, gonads, mesonephros region (AGM). Within the complex structure of the embryonic head, the localization and phenotype of HSCs and hematopoietic progenitor cells (HPCs) remains unknown. Since the Ly6a(Sca1)GFP transgene is a reporter for all AGM HSCs and some hemogenic endothelial cells (HEC), we examined these embryos for head HSC, HEC and HPC. Wholemount staining reveals some GFP+ cells in E10.5 head vasculature. By FACS analysis, 0.6% of E11.5 head cells are GFP+. To examine whether head HPC/HSC are enriched in the Ly6aGFP+ population, we sorted E11.5-E12.5 head cells and performed functional assays. Preliminary data show that head HSCs are GFP+. When we subdissected the head into forebrain (FB), midbrain (MB), and hindbrain with brachial arches (HBA) to localize HPCs, we found erythro-myeloid progenitor cells in GFP+ and GFP- fractions of all three brain regions at E10-11.5. Interestingly, in OP9 co-cutures, B cell potential was mostly observed in CD31+GFP+ of the three subregions. These results suggest that head HSCs (Ly6aGFP) are phenotypically and functionally similar to AGM HSCs. We are continuing to identify the precise head subregion(s) in which HSCs occur, examine the hemogenic potential of the head vasculature, and also the pathways affecting their development.
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