Objective:To establish a method for enriching,culturing and identifying stem-like subpopulation from human hepatic carcinoma cells,and to explore its biological properties.Methods:HepG2cells were cultured in cancer st...Objective:To establish a method for enriching,culturing and identifying stem-like subpopulation from human hepatic carcinoma cells,and to explore its biological properties.Methods:HepG2cells were cultured in cancer stem cell(CSC)medium to form spheroids.The stem-like HepG2cells obtained from tumor spheroids were then expanded.Flow cytometry was used to detect the expression of CD90 and CD133on the surface of stem-like HepG2 cells.The in vitro colony forming ability and in vivo tumorigenicity were detected by clone formation assay and tumorigenesis assay.Results:HepG2cells could grow in suspension and form spheroids in CSC medium.The stem-like cells had the ability of self-renewal and proliferation.The expression of CD90and CD133on the surface of these stem-like cells were higher than those of parental HepG2cells(P<0.01).The colony formation ability of stem-like cells was higher than that of parental HepG2cells.When injected with 1×106 cells,stemlike cells could form tumors earlier than parental cells in nude mice.The stem-like cells’tumorigenesis rate was higher and the tumor size was larger than those of parental HepG2cells(P<0.01).Conclusion:The suspension sphere culture method could enrich stem-like cells from HepG2cells.The obtained stem-like cells possessed the properties of self-renewal in vitro and tumorigenicity in vivo.展开更多
基金supported by the National Natural Science Foundation of China (No.81360347,No.81560493)Key Project of Natural Science Foundation of Guangxi (No. 2015GXNSFDA 139024)the Research Fund for the College Scientific Program of Educational Department of Guangxi Province(No.ZD2014027)
文摘Objective:To establish a method for enriching,culturing and identifying stem-like subpopulation from human hepatic carcinoma cells,and to explore its biological properties.Methods:HepG2cells were cultured in cancer stem cell(CSC)medium to form spheroids.The stem-like HepG2cells obtained from tumor spheroids were then expanded.Flow cytometry was used to detect the expression of CD90 and CD133on the surface of stem-like HepG2 cells.The in vitro colony forming ability and in vivo tumorigenicity were detected by clone formation assay and tumorigenesis assay.Results:HepG2cells could grow in suspension and form spheroids in CSC medium.The stem-like cells had the ability of self-renewal and proliferation.The expression of CD90and CD133on the surface of these stem-like cells were higher than those of parental HepG2cells(P<0.01).The colony formation ability of stem-like cells was higher than that of parental HepG2cells.When injected with 1×106 cells,stemlike cells could form tumors earlier than parental cells in nude mice.The stem-like cells’tumorigenesis rate was higher and the tumor size was larger than those of parental HepG2cells(P<0.01).Conclusion:The suspension sphere culture method could enrich stem-like cells from HepG2cells.The obtained stem-like cells possessed the properties of self-renewal in vitro and tumorigenicity in vivo.
