Isolation and characterization of cancer stem-like cells from MHCC97H Cell Lines

Abstract Objective To identify and isolate CD133 positive cancer stem-like cells (CD133 + cells) from the highly invasive human hepatocellular carcinoma cell line(MHCC97H), and examine their potential for clonogenicity and tumorigenicity. Methods CD133 + and CD133 − cells were isolated from MHCC97H cell line by magnetic bead cell sorting(MACS), and the potentials of CD133 + cells for colony formation and tumorigenicity were evaluated by soft agar cloning and tumor formation following nude mice inoculation. Results CD133 + cells represent a minority(0.5-2.0%) of the tumor cell population with a greater colony-forming efficiency and greater tumor production ability. The colony-forming efficiency of CD133 + cells in soft agar was significantly higher than CD133 − cells(36.8 ± 1.4 vs 12.9 ± 0.8, P 3 CD133 + cells, 4/5 with 1 × 10 4 CD133 + cells and 5/5 with 1 × 10 5 CD133 + cells developed detectable tumors at the injection site, while only one tumor was found in mice treated with same numbers of CD133 − cells. Conclusion CD133 may be a hallmark of liver cancer stem cells (CSC) in human hepatocellular carcinoma(HCC), because the CD133 + cells identified and isolated with anti-CD133 labeled magnetic beads from MHCC97H cell line exhibit high potentials for clonogenicity and tumorigenicity. These CD133 + cells might contribute to hepatocarcinogenesis, as well as the growth and recurrence of human HCC, and therefore may be a useful target for anti-cancer therapy.