Endothelial outgrowth cells are not derived from CD133+ cells or CD45+ hematopoietic precursors

Objective - Two types of endothelial progenitor cells ( EPCs), early EPCs and late EPCs ( also called endothelial outgrowth cells [ EOCs]), were described in vitro previously. In this report, we dissect the phenotype of the precursor(s) that generate these cell types with focus on the markers CD34, CD133, and vascular endothelial growth factor receptor- 2 ( VEGFR2) that have been used to identify putative circulating endothelial precursors. We also included CD45 in the analysis to assess the relation between CD34(+) hematopoietic progenitors ( HPC), CD34(+) endothelial precursors, and both in vitro generated EPC types. Addressing this issue might lead to a better understanding of the lineage and phenotype of the precursor( s) that give rise to both cell types in vitro and may contribute to a consensus on their flowcytometric enumeration. Methods and Results - Using cell sorting of human cord blood ( UCB) and bone marrow ( BM) cells, we demonstrate that EOC generating precursors are confined to a small CD34(+) CD45(-) cell fraction, but not to the CD34(+) CD45(-) HPC fraction, nor any other CD45(+) subpopulation. CD34(+) CD45(-) HPC generated monocytic cells that displayed characteristics typical for early EPCs. Phenotypic analysis showed that EOC generating CD34(+) CD45(-) cells express VEGFR2 but not CD133, whereas CD34(+) CD45(-) HPC express CD133 as expected, but not VEGFR2. Conclusion - EOCs are not derived from CD133(+) cells or CD45 hematopoietic precursors.