Generation of DCs from 13dpc FL-derived Lin⁻c-kit⁺ HPCs.

13 dpc FL-derived Lin⁻c-kit⁺ HPCs were cocultured with stromal cell lines PA6 or OP9 cells in the presence of GM-CSF + SCF + Flt3L for 12 to 14 days, and restimulated by GM-CSF + mTNFα for an additional 3 to 5 days. (A) A phase contrast microscopic observation was performed on cocultured FL-derived HPCs with PA6 or OP9 cells in the presence of GM-CSF + SCF + Flt3L for 12 to 14 days before and after replanted into new plates and restimulated by GM-CSF + mTNFα for an additional 3 to 5 days. Original magnification: × 200. (B) Immunophenotypic detection was performed on nonadherent cells from FL-derived HPC cultures at the indicated time points and culture conditions. These cells were sequentially stained with biotinylated CD86 MoAb and PE-labeled Ia MoAb, whereas CD86 was revealed by FITC-streptavidin. The quads were set up on the isotype-matched control dot plot. These results are representative of 3 independent experiments.