Fig. 1.

Production of cells in stromal co-cultures. Co-cultures were established by overlay of Lin⁻ bone marrow over 5G3 stroma. (a) Hematopoietic cells adhered to stroma were observed under phase contrast microscopy at day 21. Bar represents 100 μm. (b) Gating procedure to detect CD11b⁺CD11c⁺ cells is shown for cells produced in co-cultures at 28-days. (c) Cell production was assessed at 14, 21 and 28 days using antibody staining and multicolor flow cytometry. Production of the CD11b⁻CD11c⁻, CD11b⁺CD11c⁻, CD11b⁺CD11c⁺, CD11b⁺CD11c⁺MHC-II⁻(L-DC) and CD11b⁺CD11c⁺MHC-II⁺ (cDC-like) subsets was calculated in terms of proportion of each subset amongst the total CD11b⁺ and/or CD11c⁺ population. Graphs show mean ± S.E. for triplicate co-cultures. Cell production in terms of subset size is statistically different (p ≤ 0.05) for days 21 and 28 compared with day 14 in terms of increased dendritic cell and reduced myeloid cell production