Cellular heterogeneity and therapeutic response profiling of human IDH-wildtype glioma stem cell cultures

Glioblastoma stem cell cultures (GSCs) are initiated from glioblastoma (GBM) surgical resection tissue, and can capture and propagate key GBM molecular and cellular features. We have deeply characterized four IDH-wildtype GSCs from unrelated adults as cellular models of a majority of adult primary GBMs. These GSC cultures could be continuously propagated in defined, serum-free media and 5% oxygen without requiring specialized growth substrates. They had well-defined genomic and mtDNA variants and gene and protein expression profiles. All four had highly reproducible dose response curves to the GBM standard-of-care therapies of ionizing radiation (IR) and temozolomide (TMZ). Expressed lentiviral barcodes, mtDNA variants and single cell gene expression profiling were used to define cellular heterogeneity, and to track individual cell trajectories and their gene expression programs over 40 days after IR treatment. These well-characterized GSC cultures can support many high throughput in vitro assay formats, as well as xenograft, organoid and other GBM disease modeling approaches. Their use should help to better understand GBM biology, and to identify new and more effective GBM therapies and treatment regimens.