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Firenze, Italy

Stecca B.,Tumor Cell Biology Unit | Santini R.,Tumor Cell Biology Unit | Pandolfi S.,Tumor Cell Biology Unit | Penachioni J.Y.,Tumor Cell Biology Unit
Current Protocols in Stem Cell Biology | Year: 2013

Melanoma is the most aggressive skin cancer. This unit illustrates protocols for culture and isolation of human melanoma cancer stem cells/tumor-initiating cells (CSC/TIC). We describe two complementary methods to enrich for melanoma CSC/TIC. The first approach exploits the ability of CSC/TIC to growas tumor spheres in low-adherent culture conditions, as previously shown for neural stem cells and human embryonic stem cells. As a second approach, melanoma CSC/TIC are enriched by fluorescence-activated cell sorting for the aldehyde dehydrogenase (ALDH) enzyme activity.We previously showed that melanoma cells with high ALDH activity (ALDHhigh) are endowed with higher self-renewal and tumorigenic abilities than the population with low activity (ALDHlow), suggesting that ALDH might be a good marker to select for melanoma CSC/TIC. This unit will also describe how to functionally test melanoma CSC/TIC by determining selfrenewal in vitro and tumor-forming abilities in vivo using orthotopic xenograft assay. © 2013 by John Wiley & Sons, Inc. Source

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