title: Culture and isolation of melanoma-initiating cells.
abstract: 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 grow as 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 (ALDH(high)) are endowed with higher self-renewal and tumorigenic abilities than the population with low activity (ALDH(low)), 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 self-renewal in vitro and tumor-forming abilities in vivo using orthotopic xenograft assay.
authors: Tumor Cell Biology Unit, Core Research Laboratory, Istituto Toscano Tumori, Florence, Italy.
authors: Barbara
authors: B
authors: Stecca
authors: Roberta
authors: R
authors: Santini
authors: Silvia
authors: S
authors: Pandolfi
authors: Junia Y
authors: JY
authors: Penachioni
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D008545:Melanoma / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000473:pathology*; D018874:Spheroids, Cellular / Q000473:pathology; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays