Identification of neuroblastoma stem cells by characterization of side population cells in the human neuroblastoma SK-N-SH cell line

PurposeThe purpose of the study was to sort side population (SP) cells from the neuroblastoma SK-N-SH cell line and to systematically investigate whether this population has stem cell characteristics.MethodsSide population and non-SP cells were separated from the SK-N-SH cell line by flow cytometry, and their morphologic characteristics were analyzed by light and electron microscopy. We also used Western blotting to analyze marker proteins, Cell Counting Kit-8 assay for proliferative ability, series differentiation studies for differentiation properties, and Matrigel invasion study and tumorigenicity assay for malignant potential.ResultsThe SK-N-SH SP cells expressed high levels of stem cell markers, had high proliferative and malignant abilities, and had the capacity for rapid differentiation. The non-SP cells expressed differentiated cell marker proteins at high levels, had low proliferative and malignant abilities, and exhibited slow differentiation.ConclusionsThe SK-N-SH SP cells have cancer stem cell–like properties.