Brain-wide neuronal circuit connectome of human glioblastoma [bulk RNA-seq]

This study explores the circuit integration of human glioblastoma organoids (GBOs) in vivo in the adult mouse brain. Here, we performed RNA sequencing analysis of GBOs at baseline conditions and various treatment timepoints of 1 mM acetylcholine (ACh). Overall design: 3D GBOs were dissociated and reaggregated into organoids of 2000 cells in 96-well U-bottomed plates. ACh was applied to a final concentration of 1 mM for various durations prior to removal of medium and sample lysis for RNA sequencing. Libraries were prepared via a modified Smart-seq3 protocol adapted for bulk samples with sample barcodes and UMIs. This allowed for pooled samples and identification with a unique barcode per sample. Gene expression could be demultiplexed with a provided sample barcode whitelist.