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    Isolation of immortalized, INK4a/ARF-deficient cells from the subventricular zone after in utero N-ethyl-N-nitrosourea exposure

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    Authors
    Savarese, Todd M.
    Jang, Taichang
    Low, Hoi Pang
    Salmonsen, Rebecca
    Litofsky, N. Scott
    Matijasevic, Zdenka
    Ross, Alonzo H.
    Recht, Lawrence D.
    UMass Chan Affiliations
    Department of Cell Biology
    Department of Biochemistry and Molecular Pharmacology
    Department of Cancer Biology
    Departments of Neurology
    Document Type
    Journal Article
    Publication Date
    2005-01-22
    Keywords
    ADP-Ribosylation Factors
    Animals
    Brain
    Brain Neoplasms
    Cell Differentiation
    Cells, Cultured
    Cerebral Ventricles
    Corpus Striatum
    Cyclin-Dependent Kinase Inhibitor p16
    DNA Primers
    Epidermal Growth Factor
    Ethylnitrosourea
    Female
    Male
    Membrane Proteins
    Polymerase Chain Reaction
    Pregnancy
    RNA, Messenger
    Rats
    Rats, Sprague-Dawley
    Tumor Suppressor Protein p14ARF
    Cancer Biology
    Molecular and Cellular Neuroscience
    Neurology
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    Link to Full Text
    http://thejns.org/doi/full/10.3171.jns.2005.102.1.0098
    Abstract
    OBJECT: Brain tumors, including gliomas, develop several months after rats are exposed in utero to N-ethyl-N-nitroso-urea (ENU). Although pathological changes cannot be detected until these animals are several weeks old, the process that eventually leads to glioma formation must begin soon after exposure given the rapid clearance of the carcinogen and the observation that transformation of brain cells isolated soon after exposure occasionally occurs. This model can therefore potentially provide useful insights about the early events that precede overt glioma formation. The authors hypothesized that future glioma cells arise from stem/progenitor cells residing in or near the subventricular zone (SVZ) of the brain. METHODS: Cells obtained from the SVZ or corpus striatum in ENU-exposed and control rats were cultured in an epidermal growth factor (EGF)-containing, chemically defined medium. Usually, rat SVZ cells cultured in this manner (neurospheres) are nestin-positive, undifferentiated, and EGF-dependent and undergo cell senescence. Consistent with these prior observations, control SVZ cells undergo senescence by the 12th to 15th doubling (20 of 20 cultures). In contrast, three of 15 cultures of cells derived from the SVZs of individual ENU-treated rats continue to proliferate for more than 60 cell passages. Each of these nestin-expressing immortalized cell lines harbored a common homozygous deletion spanning the INK4a/ARF locus and was unable to differentiate into neural lineages after exposure to specific in vitro stimuli. Nevertheless, unlike the rat C6 glioma cell line, these immortalized cell lines demonstrate EGF dependence and low clonogenicity in soft agar and did not form tumors after intracranial transplantation. CONCLUSIONS: Data in this study indicated that immortalized cells may represent glioma precursors that reside in the area of the SVZ after ENU exposure that may serve as a reservoir for further genetic and epigenetic hits that could eventually result in a full glioma phenotype.
    Source
    J Neurosurg. 2005 Jan;102(1):98-108.
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/37643
    PubMed ID
    15658102
    Related Resources
    Link to Article in PubMed
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