Profiling Glioma Stem Cell Dynamics via 3D-Based Cell Cycle Reporter Assays
Recommended Citation
Lubanska D, Roye-Azar A, Alrashed S, Cieslukowski A, Soliman MAR, deCarvalho AC, Shamisa A, Kulkarni S, and Porter LA. Profiling Glioma Stem Cell Dynamics via 3D-Based Cell Cycle Reporter Assays. Methods Mol Biol 2025;2944:119-134.
Document Type
Article
Publication Date
6-24-2025
Publication Title
Methods in molecular biology
Abstract
Successful containment of unwanted cell cycle progression in tumors such as glioblastoma (GBM) requires targeted therapeutic approaches, which rely on understanding cell cycle dynamics in response to microenvironmental stimuli. Glioma Stem Cells (GSCs) can drive tumor initiation, recurrence, therapy resistance, and are often attributed to the heterogeneity and plasticity of GBM. In vitro models, using patient-derived GSCs, provide a life relevant tool for exploration of complex molecular mechanisms underlying the aggressive characteristics of GBM. Introduction of 3D tissue culture systems permits the study of spatial complexity of the tumor mass and enables control over diverse conditions within the surrounding microenvironment. This chapter demonstrates detailed methods to study spatiotemporal changes to the cell cycle dynamics using available fluorescent cell cycle reporter systems in combination with bioinformatics-based signal intensity and localization analysis. We present a successful approach that investigates the 3D cell cycle dynamics of GSC populations. This approach utilizes GBM neurosphere and organoid cultures, which are assessed over time and under therapeutic pressure. These models can be further explored, manipulated, and customized to serve specific experimental designs.
Medical Subject Headings
Neoplastic Stem Cells; Humans; Cell Cycle; Glioma; Brain Neoplasms; Cell Culture Techniques, Three Dimensional; Tumor Microenvironment; Cell Line, Tumor; Genes, Reporter; Glioblastoma; Spheroids, Cellular
PubMed ID
40553278
Volume
2944
First Page
119
Last Page
134
