Cholinergic neuron-to-glioblastoma synapses in a human iPSC-derived co-culture model

This study examines a co-culture model of human iPSC-derived cholinergic neurons with glioblastoma (GBM) cells derived from GBM organoids (GBOs; UP-10072). To examine transcriptional regulation of GBM cells by cholinergic neurons, we performed scRNAseq of a set of distinct conditions: 1) cholinergic neurons only; 2) GBM cells only; 3) GBM cells treated with conditioned medium from neurons; 4) direct GBM cell-cholinergic neuron co-cultures. Cells from each of the 4 conditions listed previously were dissociated from coverslips. 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.