Using this approach, we expanded colon CSCs by generating colonospheres from CRC cell line (HCT116) (Notes 4.1 and 4.2), which stably expressed exogenous NANOG (HCT116-GFP/NANOG) [3, 10] (a CSC signature), as well as from parental CRC (HCT116-GFP) cells and used them to isolate potential colorectal CSCs. This evidence concerns the gene NANOG and colorectal carcinoma.