Differentiation potential of murine CDP and MDP compared with MODP and MOP and redefinition of the human CDP. (A-D) Bulk cultures. Sorted MODP, MOP, CDP, and MDP populations were cultured at 2000 cells per well under OC differentiation conditions for 5-6 days (A-B) and under MΦ (C) or DC (D) differentiation conditions for 7-8 days. (A) Light microscopic images showing TRAP+ multinuclear OCs. (B) Total TRAP+ OC yield per well, with OCs subdivided in small (3-10 nuclei), big (10-20 nuclei), giant (20-30 nuclei), and giant (30-70 nuclei). (C) MΦ yield in numbers of live MHCII+ progeny expressing CD11b and F4/80. (D) DC yield in numbers of total live cDC (MHCII+B220−CD11c+) and pDC (MHCII+B220+CD11c+) output. Data are representative of 3 experiments. Error bars indicate standard deviations. (E-G) Clonality assays. Sorted MODP, MOP, CDP, and MDP were cultured at 5 cells per well under OC differentiation conditions (E) or at 1 cell per well under MΦ (F) or DC (G) differentiation conditions. After 5 to 12 days, cultures were analyzed. Histograms depict percentages of wells with a colony out of 48 wells seeded. Data are derived from 2 experiments. Error bars indicate standard deviations. (H-I) Redefinition of the human (h)CDP. (H) Representation of the differentiation potential and cell surface markers of the hMODP15 and the hCDP.28 (I) Flow cytometric detection of indicated markers on hMODP (red) versus hCDP (black) within the same sample. Data are representative of 3 different donors. (J) Frequency of hMODPs and hCDPs among live BM cells (mean ± SD; n = 3). (K-L) Sorted hCDPs were cultured at 1000 cells per well under OC differentiation conditions for 8-10 days and under MΦ differentiation conditions for 14 days. (K) Light microscopic images showing TRAP+ multinuclear OCs. (L) Light microscopic images showing bead phagocytosing MΦs. Data are representative of 3 experiments. *P < .05; **P < .01; ****P < .0001.
