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Flow Cytometry-based Drug Screening System for the Identification of Small Molecules That Promote Cellular Differentiation of Glioblastoma Stem Cells
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1Department of Neurological Surgery, Case Comprehensive Cancer Center,Case Western Reserve University School of Medicine, 2Department of Pathology,Johns Hopkins University, School of Medicine, 3Department of Neurological Surgery, University Hospital-Case Medical Center, Case Comprehensive Cancer Center,Case Western Reserve University School of Medicine
Chapters
- 00:05Title
- 01:17Generation of Lentiviral Transcriptional Reporter Cell Lines
- 03:47Subclone Selection, Expansion, and Validation
- 04:47Determination of GFAP:GFP Expression by Flow Cytometry
- 05:58ELDA Self-renewal Assay to Assess Clonogenic Capacity
- 07:01Drug Screen
- 08:27Results: Selection and Drug Screening of Glioblastoma Stem Cell Subclones
- 09:46Conclusion
An efficient screening protocol is presented for the identification of small molecules that promote astroglial differentiation in glioblastoma stem cells (GSCs). The assay is based on a stem cell differentiation reporter whereby the expression of the enhanced GFP (eGFP) is driven by the human GFAP promoter.
Tags
Keywords: Flow CytometryDrug ScreeningGlioblastoma Stem CellsCellular DifferentiationLentivirusReporter ConstructHigh-throughput ScreeningSmall MoleculesAstroglial DifferentiationCancer Stem CellsEhlers-Danlos GlioblastomaCell LineageBloodborne PathogensNeural Stem Cell Growth MediumPolybreneDissociation Reagent
