CD34^dim cells identified as pluripotent stem cell-derived definitive hemogenic endothelium purified using bone...

Hemogenic endothelium (HE) plays a pivotal and inevitable role in haematopoiesis and can generate all blood and endothelial lineage cells in the aorta-gonad-mesonephros of mouse embryos. Whether definitive HE can prospectively isolate pure HE from human pluripotent stem cells that can spontaneously differentiate into heterogeneous cells remains unknown. Here, we identified and validated a CD34^dim subpopulation with hemogenic potential. We also purified CD34 cells with a CXCR4^- CD73^- phenotype as a definitive HE population that generated haematopoietic stem cells and lymphocytes. The frequency of CXCR4^- CD73^- CD34^dim was evidently increased by bone morphogenetic protein 4, and purified HE cells differentiated into haematopoietic cells with myeloid and T lymphoid lineages including Vδ2+ subset of γ/δ T cells. We developed a simple method to purify HE cells that were enriched in CD34^dim cells. We uncovered an initial step in differentiating haematopoietic lineage cells that could be applied to basic and translational investigations into regenerative medicine.