Cord blood CD34⁺ cells were cultured for 6 days in the presence of GM-CSF+TNF+ SCF+sAB⁺. Cells were then collected, processed for double staining using anti–CD14-PE and anti–CD1a-FITC, and FACS-sorted into [CD14⁺CD1a⁻] and [CD14⁻CD1a⁺] populations. Sorted cells were seeded in the presence of GM-CSF+TNF at 5 × 10⁴cells/mL for 6 additional days in the presence or absence of RCC CM (10%), with a last medium change being performed at day 10. (A) Phenotype of purified populations at day 12. At day 12, cells were recovered and analyzed for CD1a, CD14, CD86, and HLA-DR expression by double-color fluorescence. (B) Function of the cells generated. At day 12, cells cultured in the presence of GM-CSF+TNF alone (medium; ▪) or in the presence of RCC CM (□) were recovered, numbered, irradiated (30 Gy), and used as stimulator cells (3.3 × 10³ cells/well) in an MLR with naive CD45R_A⁺ T cells (2 × 10⁴ cells/well).