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Updated: May 13, 2026
Isolation, Enrichment, and Maintenance of Medulloblastoma Stem Cells
Published on: September 1, 2010
Transient treatment with epigenetic modifiers yields stable neuroblastoma stem cells resembling aggressive large-cell
Naohiko Ikegaki1, Hiroyuki Shimada, Autumn M Fox
1Department of Anatomy and Cell Biology, College of Medicine, University of Illinois at Chicago, IL 60612, USA.
Abstract:
Cancer stem cells (CSCs) are plastic in nature, a characteristic that hampers cancer therapeutics. Neuroblastoma (NB) is a pediatric tumor of neural crest origin, and half of the cases are highly aggressive. By treating NB cell lines [SKNAS, SKNBE(2)C, CHP134, and SY5Y] with epigenetic modifiers for a short time, followed by sphere-forming culture conditions, we have established stem cell-like NB cells that are phenotypically stable for more than a year. These cells are characterized by their high expression of stemness factors, stem cell markers, and open chromatin structure. We referred to these cells as induced CSCs (iCSCs). SKNAS iCSC and SKNBE(2)C iCSC clones (as few as 100 cells) injected s.c. into SCID/Beige mice formed tumors, and in one case, SKNBE(2)C iCSCs metastasized to the adrenal gland, suggesting their increased metastatic potential. SKNAS iCSC xenografts showed the histologic appearance of totally undifferentiated large-cell NBs (LCNs), the most aggressive and deadly form of NB in humans. Immunohistochemical analyses showed that SKNAS iCSC xenografts expressed high levels of the stem cell marker CXCR4, whereas the SKNAS monolayer cell xenografts did not. The patterns of CXCR4 and MYC expression in SKNAS iCSC xenografts resembled those in the LCNs. The xenografts established from the NB iCSCs shared two common features: the LCN phenotype and high-level MYC/MYCN expression. These observations suggest both that NB cells with large and vesicular nuclei, representing their open chromatin structure, are indicative of stem cell-like tumor cells and that epigenetic changes may have contributed to the development of these most malignant NB cells.
Insights
Induced cancer stem cells (iCSCs) were created from neuroblastoma cells using epigenetic modifiers. These iCSCs formed aggressive tumors in mice, highlighting their potential role in advanced neuroblastoma.
Area of Science:
- Developmental Biology
- Cancer Research
- Epigenetics
Background:
- Cancer stem cells (CSCs) exhibit plasticity, complicating cancer treatment strategies.
- Neuroblastoma (NB), a pediatric cancer of neural crest origin, frequently presents as highly aggressive.
Purpose of the Study:
- To establish stable, stem cell-like neuroblastoma cells (iCSCs) using epigenetic modification.
- To characterize the stemness, tumorigenic, and metastatic potential of these induced CSCs (iCSCs).
Main Methods:
- Neuroblastoma cell lines were treated with epigenetic modifiers and cultured under sphere-forming conditions.
- Induced CSCs (iCSCs) were characterized by stemness factors, stem cell markers, and chromatin structure.
- Tumorigenicity and metastatic potential were assessed by xenotransplantation into SCID/Beige mice.
Main Results:
- Phenotypically stable iCSCs were generated, exhibiting high stemness factor expression and open chromatin.
- iCSC xenografts formed tumors, with one instance showing adrenal gland metastasis, indicating enhanced metastatic potential.
- Xenografts displayed features of aggressive large-cell neuroblastomas (LCNs), with high expression of CXCR4 and MYC/MYCN.
Conclusions:
- Epigenetic modification can induce stable, stem cell-like properties in neuroblastoma cells.
- These induced CSCs (iCSCs) possess tumorigenic and metastatic potential, mirroring aggressive neuroblastoma phenotypes.
- Open chromatin structure and high MYC/MYCN expression in NB cells may indicate malignancy and stem-like properties.
