Identification of a massive reserve of hematopoietic progenitors in mice

Previous studies have demonstrated that mice null (-/-) for either CD34 or c-mpl are viable and have greatly decreased numbers of multipotential (CFU-Mix), erythroid (BFU-E), and granulocyte-macrophage (CFU-GM) progenitor cells in the bone marrow (BM), spleen (Spl) and peripheral blood (PB), without noticeable decreases in the nucleated cellularity of these organs. To evaluate the significance of these two proteins further, mice null for both CD34 and c-mpl were assessed for hematopoietic progenitor cells (HPC) and nucleated cellularity and compared with these cells in CD34(-/-) and c-mpf(-/-) mice. The following progenitors were assessed: CFU-GM, BFU-E, CFU-Mix with an erythroid component, CFU-Mix with erythroid and megakaryocyte components, nonerythroid CFU with a megakaryocyte (Meg) component and pure CFU-Meg. Results demonstrated significant decreases in progenitors in the BM of dual CD34/c-mpl(-/-) mice compared to decreases from CD34(-/-) or c-mpl(-/-) mice; progenitor numbers in CD34/c-mpl(-/-) mice were decreased by 83-99.3 % compared to that in wild-type littermate control mice. Decreases in progenitors in spleens of c-mpl(-/-) mice (89-96 %) were more drastic than those of CD34-/- mice (50-78 %) whereas those of dual CD34/c-mpl(-/-) mice were equal to or lower than that of c-mpl(-/-) mice (93-98 %). Decreases in PB progenitors were seen in the c-mpl(-/-) and dual CD34/c-mpl(-/-) mice (75-90 %). Whereas progenitor cells in BM, Spl and PB were drastically reduced in dual CD34/c-mpl(-/-) mice compared to controls, absolute numbers of nucleated cells in these organs were essentially not reduced. These studies demonstrate that CD34 and c-mpl have non-redundant effects on maintenance of steady-state hematopoiesis and highlight how few progenitor cells are required in steady-state conditions to populate and maintain the BM, Spl, and PB with nucleated cells.