CD109 is expressed on a subpopulation of CD34(+) cells enriched in hematopoietic stem and progenitor cells

CD109 is a monomeric cell surface glycoprotein of 170 kD that is expressed on endothelial cells, activated but not resting T-lymphocytes, activated bu t not resting platelets, leukemic megakaryoblasts, and a subpopulation of b one marrow CD34(+) cells. Observing an apparent association between CD109 e xpression and the megakaryocyte lineage (MK), we sought to determine whethe r CD109 was expressed on MK progenitors. In fetal bone marrow (FBM), a rich source of MK progenitors, CD109 is expressed on a mean of 11% of CD34(+) c ells. Fluorescence activated cell sorting (FACS) of FBM CD34(+) cells into CD109(+) and CD109(-) fractions revealed that the CD34(+)CD109(+) subset co ntained virtually all assayable MK progenitors, including the colony-formin g unit-MK (CFU-MK) and the more primitive burst-forming unit-MK (BFU-MK). T he CD34(+)CD109(+) subset also contained all the assayable burst-forming un its-erythroid (BFU-E), 90% of the colony-forming units-granulocyte/macropha ge (CFU-GM), and all of the more primitive mixed lineage colony-forming uni ts (CFU-mix). In contrast, phenotypic analysis of the CD34(+)CD109(-) cells in FBM, adult bone marrow (ABM) and cytokine-mobilized peripheral blood (M PB) demonstrated that this subset comprises lymphoid-committed progenitors, predominantly of the B-cell lineage. CD109 was expressed on the brightest CD34(+) cells identifiable not only in FBM, but also in ABM and MPB indicat ing that the most primitive, candidate hematopoietic stem cells (HSC) might also be contained in the CD109(+) subset. In long-term marrow cultures of FBM CD34(+) cells, all assayable cobblestone area forming cell (CAFC) activ ity was contained within the CD109(+) cell subset. Further phenotypic analy sis of the CD34(+)CD109(+) fraction in ABM indicated that this subset inclu ded candidate HSCs that stain poorly with CD38, but express Thy-1 (CD90) an d AC133 antigens, and efflux the mitochondrial dye Rhodamine 123 (Rho123). When selected CD34(+) cells were sorted for CD109 expression and Rho123 sta ining, virtually all CAFC activity was found in the CD109(+) fraction that stained most poorly with Rho123. CD34(+) cells were also sorted into Thy-1( +)CD109(+) and Thy-1(-)CD109(+) fractions and virtually all the CAFC activi ty was found in the Thy1(+)CD109(+) subset. In contrast, the Thy-1(-)CD109( +) fraction contained most of the short-term colony-forming cell (CFC) acti vity. CD109, therefore, is an antigen expressed on a subset of CD34(+) cell s that includes pluripotent HSCs as well as all classes of MK and myeloeryt hroid progenitors. In combination with Thy-1, CD109 can be used to identify and separate myelo-erythroid and all classes of MR progenitors from candid ate HSCs. (C) 1999 International Society for Experimental Hematology. Publi shed by Elsevier Science Inc.