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Colorectal cancer (CRC) is the 2nd highest cause of cancer death in the United States. Increasing evidence suggests that a small population of cancer-initiating cells (CICs) or cancer stem cells (CSCs) are responsible for tumorigenesis and the exceptional drug resistance to anti-cancer therapies., CR4 is a novel CIC-enriched, highly tumorigenic, clonogenic and 3D spheroid-forming colon cancer cell line derived from the liver metastasis of a male patient with stage 4 colon cancer (1). Primary tumor cells were established on type I collagen coated flasks in serum-free stem cell media (1). Holoclones, small stem-like cells that are densely packed, were selectively subcloned from the heterogeneous culture and propagated. Cells were then passaged as 3D spheroids and in NOD/SCID mice xenografts leading to the eventual establishment of the CR4 human colorectal cancer cell line. Majority of CR4 cells express high but variable levels of common cancer stem cell markers including CD44, CD166, EpCAM, and Lgr51. CR4 cell line has the G12V mutation on KRAS exon 2 (Botchkina GI, communications). Reference:1. Rowehl RA, Burke S, Bialkowska AB, Pettet DW III, Rowehl L, Li E, Antoniou E, Zhang Y, Bergamaschi R, Shroyer KR, Ojima I., Botchkina GI (2014) PLOS 9(6): e99091.