Derivation and lineage potential of ivGMPs. (A) GMPs or preGMPs isolated from WT BM were cultured in presence of 100 ng/mL SCF and Flt3L for 3 days, and then analyzed for CD16/32 and CD11b expression via flow cytometry. Representative FACS plots are shown. (B) Freshly isolated GMPs and ivGMPs were cultured in either SCF, IL-3, TPO, EPO, 10% FCS or CSF-1, G-CSF, and 10% FCS, and analyzed for M (CD11b^pos F4/80^pos Ly6G^neg) and G (CD11b^pos F4/80^neg Ly6G^pos) output via flow cytometry after 4 to 5 days (n = 3 to 6). (C) Freshly isolated GMPs and ivGMPs were cultured in methylcellulose containing SCF, IL-3, IL-6, and EPO (M3434). After 5 to 7 days, colonies were enumerated and identified according to morphology (clonogenicity WT GMP = 48% ± 7.5% SD; ivGMPs = 45% ± 6% SD; n = 3 experiments). No significant difference was detected. EPO, erythropoietin; SD, standard deviation; TPO, thrombopoietin.