title: Isolation and identification of cancer stem-like cells from murine melanoma cell lines.
abstract: In current study, cancer stem-like cells in the murine melanoma B16F10 cells were investigated. CD phenotypes of the B16F10 cells were analyzed by flow cytometry, and the specific CD phenotype cells from the B16F10 cells were isolated by MACS. Then we used colony formation assay in soft agar media, the cell growth assay in serum-free culture media as well as the tumorigenicity investigation of the specific CD phenotype cells in C57BL/6 mice, respectively, to identify cancer stem-like cells in the B16F10 cells. The results showed that the B16F10 cells could form spherical clones in serum-free culture media, and the rate of clonegenesis of CD133+, CD44+ and CD44+CD133+ cells was higher than that of CD133-, CD44- and CD44+CD133- cells in soft agar media, respectively. The tumorigenic potential of CD133+, CD44+, CD44+CD133+ cells and CD44+CD133+CD24+ cells was stronger than that of CD133-, CD44-, CD44+CD133- cells and CD44+CD133+CD24- cells in mice, respectively. In conclusion, the CD44+CD133+CD24+ cells have some biological properties of cancer stem-like cells or are highly similar to the characteristics of cancer stem cells (CSC). These results provide an important method for identifying cancer stem-like cells in B16F10 cells and for further cancer target therapy.
authors.affiliation: Department of Pathogenic Biology and Immunology, School of Basic Medical Science, Southeast University, Nanjing 210009, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Dou
authors.forename: Meng
authors.initials: M
authors.lastname: Pan
authors.forename: Ping
authors.initials: P
authors.lastname: Wen
authors.forename: Yating
authors.initials: Y
authors.lastname: Li
authors.forename: Quan
authors.initials: Q
authors.lastname: Tang
authors.forename: Lili
authors.initials: L
authors.lastname: Chu
authors.forename: Fengshu
authors.initials: F
authors.lastname: Zhao
authors.forename: Chuilian
authors.initials: C
authors.lastname: Jiang
authors.forename: Weihua
authors.initials: W
authors.lastname: Hu
authors.forename: Kai
authors.initials: K
authors.lastname: Hu
authors.forename: Ning
authors.initials: N
authors.lastname: Gu
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000276:immunology; D051927:CD24 Antigen / Q000276:immunology; D045744:Cell Line, Tumor; D002469:Cell Separation; D002999:Clone Cells; D006023:Glycoproteins / Q000276:immunology; D018960:Hyaluronan Receptors / Q000276:immunology; D016130:Immunophenotyping; D008545:Melanoma / Q000276:immunology / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009374:Neoplasms, Experimental / Q000473:pathology; D014411:Neoplastic Stem Cells / Q000276:immunology / Q000473:pathology*; D010455:Peptides / Q000276:immunology
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D051927:CD24 Antigen; D006023:Glycoproteins; D018960:Hyaluronan Receptors; D010455:Peptides; C000606024:Prom1 protein, mouse