Cells were indicated with 10 ng/mL of human recombinant basic fibroblast growth factor (R&D Systems, Minneapolis, MN) and 20 ng/mL of epidermal growth factor (R&D Systems) in serum-free Dulbecco's Modified Eagle's Medium-F12 (Gibco) medium. Spheres were observed using a microscope (Olympus, Tokyo, Japan).

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