Flow cytometric analysis of day-9 cultured cord blood erythroid cells. Cord blood CD34+ cells were cultured for 9 days under erythroid conditions as described. These cells were labeled with MoAbs to the erythroid-specific markers, glycophorin C (A through C), glycophorin A (D through F), or band III (G through I) or with the CD164(103B2/9E10) MoAb (J through L), followed by FITC anti-mouse IgG. Cells were analyzed on a FACSCalibur on the basis of forward and side light scatter parameters. Either the whole cell population was analyzed (dark histograms A, D, G, J) or the cells were gated into two subsets based, both with low side scatter and with low to medium forward scatter (R1) or medium to high forward scatter (R2) before fluorescence analysis (dark histograms B, E, H, K for R1 and C, F, I, L for R2). Values adjacent to each histogram represent median fluorescence values. Isotype-matched negative controls for A through I were an irrelevant IgG1 mouse MoAb (light histograms in A through C) or an irrelevant mouse IgG3 MoAb (light histograms in J through L). Median fluorescence values for negative controls were A = 9; B = 9; C = 6; J = 9; K = 9; L = 8.
