Abstract: Glioblastoma (GBM) stem-like cells (GSCs) represent the most undifferentiated state of malignant cells with distinct biological characteristics. The fraction of these cells within a glial brain tumor, ranging from 2–30%, is correlating with the increasingWHOstage and poor prognosis of patients' survival. GSCsrepresent the least vulnerable, thus most preferential target cell population to be exposed to various therapeutic modalities, although the underlying mechanisms of this resistance are not yet fully understood. For the development of GSC-targeting therapies, further in depth studies are needed using enriched and stable GSCcell populations. Here, we discuss the current approaches ofGSCisolation and validation based on expression of stemness and oncogenic markers as well as on functional assays. The enrichment of GSC phenotypes in established cell lines and/or primary tumor cultures, achieved by different strategies, is reviewed, providing a comprehensive comparison of selected studies and contemplating the characterization of the plethora of variants of reported GBM population exhibiting the GSC phenotype.
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