Sorting of CD34⁻c-kit^−/lowCD15⁺ cells grown by SCF + TPO from CD34⁺cord blood cells. The cultured cells grown by SCF + TPO at 3 weeks were stained with APC-conjugated anti-CD34 MoAb, PE-conjugated anti–c-kit MoAb and FITC-conjugated anti-CD15 MoAb. As negative controls, APC-, PE-, and FITC-conjugated mouse isotype-matched Ig were used. (A) The viable cell region (R1) was gated on the basis of FSC and SSC. (B) CD34 and CD15 expressions of the cells in the R1 region. (C) The gate (R2) was set on CD34⁻ cells. (D) The expressions of c-kit and CD15 on these cells were then examined. The cells in the R3 region were sorted as CD34⁻c-kit^−/low CD15⁺cells.