Figure 3

Ex vivo generation of functional DCs. (A) Schematic illustration of expansion of human and mouse DCPs in culture. The human (CD34⁺) or mouse (Sca1⁺/Lin^-) HPCs were cultured on LSC-KFT63b for 10 days, and then transferred to LSC-KFT-GM15 (LSC-KFT-mGM15 for mouse cells) to expand for 30 days. The resulting DCPs were further cultured in medium supplemented with GM-CSF, IL-15 and growth factors to induce immature and mature DCs. The average fold of expansion in each stage is indicated. (B) Kinetic analysis of myeloid cell differentiation markers. The expression kinetics of molecular markers for myeloid cells including PU.1, Langerin, Id2, hIL7R-α, CCL17, hCCR6, and E-cadherin (E-CAD) in the developing human DCPs were examined by RT-PCR. (C) Expression kinetics of DC surface markers of the DCPs based on flow cytometry analysis. (D) Surface phenotype of day 37 DCPs from the LSC-KFT-GM15 culture. The number inside each of the flow graph represents percentage of positive cells.