Isolation Of CD133~+ Cell And Stem Cell Property From Hepatocarcinoma Hep3B Cell

AIM:To detect the expression of CD133 in hepatocellular carcinoma Hep3B cell line, and to detect proliferative and self-renew ability of CD133~+ tumor cells in vitro, implantation experiment of CD133~+ 3B cell in vivio, and study its stem-cell property.METHODS:Flow cytometry was used to detect the expression of putative tumor-initiated cell marker CD133 in Hep3B cell line and the isolation technique with immunomagnetic beads was applied to purify CD133~+ cells. The proliferative ability of CD133~+ tumor cells was estimated by MTT assay in vitro, and the proliferative ability of CD133~+, CD133- and control Hep3B cells were statistically compared.CD133~+ tumor cells were cultured in serum-free medium after isolation and observed. Expression of CD133~+ cells in sphere cells of CD133~+ were detected by flow cytometry.Tumor cells implantation experiments were done to test the tumorigenicity of CD133~+, CD133- and control Hep3B cells.RESULTS: 9.13% CD133~+ cells were detected in Hep3B cell line. In serum-free RPMI 1640, their UV optical of the CD133~+ tumor cells was 0.311, 0.504, 0.602 and 0.800 at 1, 3, 5 and 7 days respectively. Compared with CD133~- cells and control Hep3B cells, CD133~+ cells demonstrated increased proliferation capacity.49.48% CD133~+ cells were detected insphere cells of CD133~+ cells.Tumor cell implantation experiments showed that only 1×10~3 CD133~+ cells were sufficient for tumor formation, whereas an injection of 1×10~5 CD133- cells could initiate tumors.CONCLUSION: CD133~+ cells subpopulation has stronger ability to proliferate in vitro and to form tumors in vivo than other subpopulations in Hep3B cell line. In one word, CD133~+ cell subpopulation has stem-cell property.