Isolation and identification of tumor stem cells from human ovarian carcinoma cell line A2780

OBJECTIVE: To isolate and cultivate ovarian tumor stem cells from human ovarian tumor cell line A2780,and identify their biological properties.METHODS: Cells A2780 were dissociated into single tumor cells,suspended in specific serum-free medium(SFM) to acquire ovarian tumor stem cell spheres.The expression of cell surface markers CD44 and CD133 was detected with flow-cytometry.Transwell invasive method was utilized to analyze the invasive ability in vitro and CCK-8 assays were performed to detect their proliferating potentiality.To compare their doubling time,and observe the inducing differentiation and cytologic and morphologic characteristics of the stem cell spheres.RESULTS: Nonadherent and serial-propagating tumor stem cells could be obtained from cell line A2780,which highly expressed stem cell markers CD44 and CD133(P0.05),with stronger invasive ability in vitro(t=9.354,P0.05),stronger propagating capability(t=13.682,P0.05)and shorter doubling time(t=3.773,P0.05).They could differentiate normal tumor cells in serum-supplemented medium condition.CONCLUSION: Ovarian tumor stem cell spheres can be obtained by serum-free suspended culture,which is enriched with this kind of stem cells.