Lineage potential of CD45RA^intand CD45RA^hicells. (A) CFU-C generation from 500 plated, day 21–sorted CD45RA^int and CD45RA^hi cells, grown on OP9-DL4^low (first panel) and/or OP9-Ctrl (second panel). CFU-C generation from ex vivo–derived Stem, MPP, and CD45RA^intRho^low/int CD133⁺CD7⁻CD10⁻ (ex vivo CD45RA^int) (third panel). Mean and SEM were calculated from 6 independent experiments. (B) CFU-C generation from 1000 plated ex vivo CD90⁻ (MPP), CD45RA^intRho^int cells or CD45RA^intRho^low cells. (C) Purity reanalysis of CD45RA^int day 21–sorted fractions and ex vivo CD45RA^int is shown as day 21 (+0d) (first column). Generation of CD5⁺, CD7⁺, and CD1a⁺ cells is shown in the middle columns and CD19⁺ cells vs CD33⁺ in the last column for CD45RA^int derived from OP9-Ctrl and /or DL4^low and ex vivo CD45RA^int. (D) Purity reanalysis of CD45RA^hi day 21–sorted fractions is shown as day 21 (+0d) (first column). Generation of T cells in vitro by CD45RA^hi cells derived from OP9-Ctrl or OP9-DL4^low cultures is shown in the middle columns, and generation of CD19⁺ or CD33⁺ cells is shown in the last column. Numbers within each plot indicate the percentage of cells in the indicated gates. This is a representative experiment of at least 10 independent experiments. (E) Ex vivo CD90⁻ (MPP), CD45RA^intRho^int cells or CD45RA^intRho^low cells were cultured on OP9-Ctrl for 26 days and analyzed for the expression of CD34, CD10, and CD19 by flow cytometry. Numbers within each plot indicate the percentage of cells in the indicated gates.