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Cell Biol Int. 2009 Aug;33(8):807-15. doi: 10.1016/j.cellbi.2009.05.003. Epub 2009 May 27.

Identifying tumor stem-like cells in mouse melanoma cell lines by analyzing the characteristics of side population cells.

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Department of Pathogenic Biology and Immunology, School of Basic Medical Science, Southeast University, 87# Dingjiaqiao Rd., Nanjing 210009, Jiangsu, China.


Increasingly more evidence shows that TSCs possess the characteristics of stem-like cells. However, a link between stem cells and TSCs remains to be shown. We have sorted SP cells and non-SP cells from the B16F10 cell lines by FACS, and then studied their cellular biological characteristics by using a SFCM culture method, proliferative assay in vitro, clone formation assays in soft agar and normal media, tumorigenic assays in C57BL/6 mice, and resistance to chemotherapy assay in vitro, the quantitative detecting expression of ABCG2 and their CD phenotype analysis by a FCM. We detected 0.96% SP cells in the B16F10 cells and found that they had obvious differences in characteristics from non-SP cells. They possessed a marked capacity for self-renewal in soft agar and culture medium, strong tumorigenic potential in C57BL/6 mice, apparent resistance to vinblastin in vitro, upregulated ABCG2 expression, and a high expression of CD44+CD133+CD24+ phenotypes. We conclude that there were a few of SP cells that had the characteristics of tumor stem-like cells which may provide a useful tool and a readily accessible source for further study when specific TSCs markers are unknown.

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