Culture and Isolation of Melanoma‐Initiating Cells. (1st February 2013)

• Record Type: Journal Article
• Authors: Stecca, Barbara; Santini, Roberta; Pandolfi, Silvia; Penachioni, Junia Y.
• Editors: Bhatia, Mick; Elefanty, Andrew G.; Fisher, Susan J.; Patient, Roger; Schlaeger, Thorsten; Snyder, Evan Y.
• Is Part Of: Current protocols in stem cell biology. Volume 24(2013)
• Page Start: 3.6.1
• Page End: 3.6.12
• Publication Date: 2013-02-01
• Subjects: melanoma -- cancer stem cells -- in vitro sphere assay -- in vivo xenograft assay -- Aldefluor assay -- tumor spheres; Stem cells -- Laboratory manuals; Stem cells -- Physiology; Stem Cells; Stem cells; Laboratory Manual; Laboratory manuals; 571.5
• DOI: 10.1002/9780470151808.sc0306s24
• Languages: English
• ISSNs: 1941-7322

Abstract

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. Curr. Protoc. Stem Cell Biol . 24:3.6.1‐3.6.12. © 2013 by John Wiley & Sons, Inc.