EGF and hepatocellular carcinoma: Following the generation of LCSC spheroids, we modified their environment from the stem cell-permissive medium (DMEM/F12 supplemented 1× B27, 20 ng/ml bFGF, 20 ng/ml EGF, 25ug/ml insulin) to a general HCC culture medium (DMEM-high glucose supplemented with 10% FBS) to make CD133-negative HCC populations, which were differentiated from LCSC spheroids.