Journal
BIOLOGY OPEN
Volume 10, Issue 2, Pages -Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/bio.056416
Keywords
Neural stem cells; Glioblastoma; Organoid; Neurogenesis; 3D culture; Brain tumor
Categories
Funding
- Rita Levi Montalcini funding from MIUR (Italian Ministry of Education, University and Research)
- Society for Developmental Biology (SDB) Innovation Award
- Royal Scientific Society E.P. Abraham Research Professorship [RP\R1\180165]
- Wellcome Trust [098357/Z/12/Z]
- Cancer Research UK Programme Grant [RG91505]
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The emerging 3D cultures of glioblastoma are powerful models for studying cell behavior and interactions. GSCs can efficiently integrate into brain organoids in co-culture conditions, paving the way for analyzing fate behavior and lineage progression. This establishes the 3D culture system as a viable and versatile GBM model for investigating tumor cell biology and GSC heterogeneity.
Emerging three-dimensional (3D) cultures of glioblastoma are becoming powerful models to study glioblastoma stem cell behavior and the impact of cell-cell and cell-microenvironment interactions on tumor growth and invasion. Here we describe a method for culturing human glioblastoma stem cells (GSCs) in 3D by co-culturing them with pluripotent stem cell-derived brain organoids. This requires multiple coordinated steps, including the generation of cerebral organoids, and the growth and fluorescence tagging of GSCs. We highlight how to recognize optimal organoid generation and how to efficiently mark GSCs, before describing optimized co-culture conditions. We show that GSCs can efficiently integrate into brain organoids and maintain a significant degree of cell fate heterogeneity, paving the way for the analysis of GSC fate behavior and lineage progression. These results establish the 3D culture system as a viable and versatile GBM model for investigating tumor cell biology and GSC heterogeneity.
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