Dissociated human cerebral organoids on micro-electrode arrays develop neuronal networks

Microelectrode arrays (MEAs) are increasingly used to profile the development of synchronised network activity in neural organoids, yet no organoid study has investigated the consistency of electrophysiological development across multiple cell lines. Here, we used dissociated neural organoids derived from six cell lines on MEAs to characterise functional synapse development using multiple parameters across time. The dissociated organoids had increasing functional connectivity and network activity over time across all cell lines and plasticity in response to synaptic-like stimulation. Like the organoids they were derived from, dissociated organoid cultures contained a diverse mixture of cell types, including neurons and glia. Variability in activity parameters was associated with differences in cell type composition and regional identity, which in turn were affected by donor cell line and batch effects. Cell type composition and regional identity in dissociated organoid cultures grown on MEAs was investigated using bulk RNA-seq. Overall design: Bulk RNA-seq profiling of control dissociated cerebral organoid cultures at 37 days post dissociation (82-84 days in vitro including culture time as 3D organoids) across six donor cell lines: KOLF2.1J, SCTi003-A, H1, H9, 59XY and 272XX. Two independent batches are included for the H9, KOLF2.1J and SCTi003-A lines. Three batches are included for 59XY and 272XX. Each sample represents RNA pooled from six wells.