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


METHODS
Side 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.


RESULTS
The 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.


CONCLUSIONS
The SK-N-SH SP cells have cancer stem cell-like properties.
Department of Surgery, Children's Hospital of Fudan University, Shanghai 201102, China.
Shiqin
S
Qi
Jicui
J
Zheng
Haitao
H
Zhu
Lin
L
Yang
Xianmin
X
Xiao
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D014408:Biomarkers, Tumor / Q000032:analysis; D045744:Cell Line, Tumor / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure; D002469:Cell Separation / Q000379:methods*; D003094:Collagen; D004338:Drug Combinations; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis; D007797:Laminin; D008562:Membrane Glycoproteins / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008854:Microscopy, Electron; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000032:analysis; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D009419:Nerve Tissue Proteins / Q000032:analysis; D009447:Neuroblastoma / Q000737:chemistry / Q000473:pathology* / Q000648:ultrastructure; D064531:Peripherins; D011509:Proteoglycans; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis; D009418:S100 Proteins / Q000032:analysis; D014410:Tumor Stem Cell Assay
D015415:Biomarkers; D014408:Biomarkers, Tumor; D004338:Drug Combinations; D007381:Intermediate Filament Proteins; D007797:Laminin; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D064531:Peripherins; D011509:Proteoglycans; D009418:S100 Proteins; C059485:matrigel; D003094:Collagen; D019009:Proto-Oncogene Proteins c-kit