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Nano-liposome was used to load Salinomycin(SAL) and Doxorubicin(DOX).Four kinds of liver cell lines were selected and experimental performance including cell culture, passage, cryopreservation and thawing were mastered.HepG2 tumor sphere (HepG2-TS) was screened as liver cancer stem cells by culturing HepG2 cells in serum-free medium.In vitro cytotoxicity-test shows that SAL-Na could sensitize DOX in liver cancer cells_and the ideal proportions for synergic effect of DOX and SAL-Na were defined as 1∶1 and 1∶5.