Brain-wide neuronal circuit connectome of human glioblastoma

This study explores the circuit integration of human glioblastoma organoids (GBOs) in vivo in the adult mouse brain. We performed single cell RNA sequencing (scRNA-seq) to understand the cell state diversity of malignant tumor cells in GBOs at baseline and after stimulation by 1 mM acetylcholine (ACh) for 1 hour. Sliced neocortical organoids (SNOs) were also sequenced to study gene expression properties of neural stem cells (NSCs). 3D GBOs were either not treated or treated with 1 mM ACh for 1 hour, upon which they were dissociated to single cells and sorted into lysis-buffer containing 96-well plates. 3D SNOs were processed the same way without treatment. Whole cell scRNAseq libraries were performed via a modified Smart-seq3 protocol with cell barcodes. Therefore, cells could be pooled together and identified with a unique barcode. Gene expression data could be demultiplexed with a provided barcode whitelist.