Isolation And Identification Of Tumor Stem Cells From Human Ovarian Carcinoma Cell Line A2780

Objective:The tumor stem cells have the ability of unlimited proliferation and multiple proliferation, have characteristics such as high tumorigenicity and radiochemotherapy resistance, which are responsible for the relapse and metastasis of the carcinoma. The features of the ovarian cancer are drug-resistant and relapsing, which is highly indicative of existence of tumor stem cell in it. In this study, we prepare to isolate and cultivate ovarian tumor stem cells from human ovarian tumor cell line A2780 and to identify their biological properties.Methods:Cells A2780 were dissociated into single tumor cells, suspended in specific serum-free medium(SFM), in order to acquire ovarian tumor stem cell spheres. The expression of stem 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. Statistic analysis:T-test andχ2-test were performed to analyse the measurement data and enumeration data by SPSS software(version 17.0).Results:The ovarian cell line A2780 can survive and proliferate in SFM, nonadherent and serial-propagating tumor stem cells could be obtained, they highly expressed stem cell markers CD44 and CD133 (P<0.05),own stronger invasiveness(t=9.354),had stronger propagating capability(t=13.682, P<0.05) and shorter doubling time(t=3.773,P<0.05). 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 enriched this kind of stem cells.