Titlebook: Glioblastoma; Methods and Protocol Dimitris G.‘Placantonakis Book 2018 Springer Science+Business Media, LLC 2018 xenografts.GBM cancer stem

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Whole Genome Sequencing-Based Discovery of Structural Variants in Glioblastoma,ent of disease. The goal is to use high-throughput sequencing to identify specific variants that drive tumorigenesis within each individual’s tumor genomic profile. The significance of copy number and structural variants in glioblastoma makes it essential to broaden the search beyond oncogenic singl

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Whole Genome DNA Methylation Analysis of Human Glioblastoma Using Illumina BeadArrays, glioblastoma molecular profiles. The procedure spans four days, and can be performed by a single laboratory technician. Starting with as little as 250 ng of DNA input, this method allows the flexibility to begin with DNA derived from either formalin-fixed, paraffin-embedded (FFPE) or fresh tissue a

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Establishing Primary Human Glioblastoma Tumorsphere Cultures from Operative Specimens, or by tumorsphere culture in suspension. Here, we provide a detailed protocol for establishing patient-derived tumorsphere cultures. Such cultures are enriched for GBM stem cells (GSCs) and can be used to generate orthotopic tumor xenografts in the brain of immunocompromised mice. We also point out

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Isolation of Glioblastoma Stem Cells with Flow Cytometry,cell sorting (FACS) using CD133 as a surface marker. The use of a directly conjugated antibody to an APC fluorophore against the CD133 molecule provides sufficient and clear detection of positive cells from the rest of the population. This strategy avoids an unnecessary secondary antibody incubation

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Lentiviral Transduction of Primary Human Glioblastoma Cultures, cultures. Lentiviruses stably transduce both dividing and non-dividing cells, such as quiescent cancer stem cell populations. The viral envelope is pseudotyped with the vesicular stomatitis virus envelope glycoprotein G (VSV-G), which renders the lentiviral particles pantropic, so that they can inf

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Selective Targeting of CD133-Expressing Glioblastoma Stem Cells Using Lentiviral Vectors,or progression and recurrence, by virtue of their robust tumor-propagating potential and resistance to conventional chemoradiotherapy. Therefore, targeting GSCs holds significant therapeutic promise. Expression of CD133 (PROM1), a cell surface glycoprotein, has been associated with the GSC phenotype