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: Barbara Stecca, Roberta Santini, Silvia Pandolfi, Junia Y Penachioni
• Journal: Current protocols in stem cell biology (Curr Protoc Stem Cell Biol) Vol. Chapter 3 Pg. Unit 3.6 (Feb 2013) ISSN: 1938-8969 [Electronic] United States
• PMID: 23404674 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Topics

• Animals
• Cell Proliferation
• Cell Separation (methods)
• Humans
• Melanoma (pathology)
• Mice
• Mice, Nude
• Neoplastic Stem Cells (pathology)
• Spheroids, Cellular (pathology)
• Tumor Cells, Cultured
• Xenograft Model Antitumor Assays