title: Isolation of Circulating Melanoma Cells.
abstract: Circulating melanoma cells (CMCs) represent critical mediators of metastatic melanoma progression. However, isolation and characterization of CMCs has been challenging due to the low frequency of these cells and the paucity of melanoma-specific cell surface markers. Herein, we describe a method for the isolation of CMCs that employs two independent markers, displays high sensitivity for CMC enrichment, and can be readily adapted to include additional molecular melanoma markers of interest. CMCs isolated by this method are enriched for ABCB5-positive melanoma stem cells, are tumorigenic in xenotransplantation assays, and can be used for phenotypical, genetic, and functional investigations of CMC biology.
authors.affiliation: Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.|Transplantation Research Program, Division of Nephrology, Children's Hospital Boston, Harvard Medical School, 300 Longwood Avenue, Boston, MA, 02115, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Ma
authors.affiliation: Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. markus.frank@childrens.harvard.edu.|Transplantation Research Program, Division of Nephrology, Children's Hospital Boston, Harvard Medical School, 300 Longwood Avenue, Boston, MA, 02115, USA. markus.frank@childrens.harvard.edu.
authors.forename: Markus H
authors.initials: MH
authors.lastname: Frank
keywords: ABCB5; Circulating melanoma cells; Circulating tumor cells; Flow cytometry; Melanoma stem cells; Metastasis; Tumorigenicity; Xenotransplantation