Electrophysiological development and functional plasticity in dissociated human cerebral organoids across multiple cell lines

Adam Pavlinek; Sara Guerrisi; Kara O'Driscoll; Lucia Dutan Polit; Roland Nagy; APEX consortium; Madeline A Lancaster; Anthony C Vernon; Deepak P Srivastava

doi:10.1016/j.crmeth.2026.101371

Electrophysiological development and functional plasticity in dissociated human cerebral organoids across multiple cell lines

Cell Rep Methods. 2026 Mar 27:101371. doi: 10.1016/j.crmeth.2026.101371. Online ahead of print.

Authors

Adam Pavlinek¹, Sara Guerrisi², Kara O'Driscoll², Lucia Dutan Polit², Roland Nagy²; APEX consortium; Madeline A Lancaster³, Anthony C Vernon², Deepak P Srivastava⁴

Affiliations

¹ Department of Basic and Clinical Neuroscience, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London SE5 9RT, UK; MRC Centre for Neurodevelopmental Disorders, King's College London, London, UK. Electronic address: ap2025@cantab.ac.uk.
² Department of Basic and Clinical Neuroscience, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London SE5 9RT, UK; MRC Centre for Neurodevelopmental Disorders, King's College London, London, UK.
³ MRC Laboratory of Molecular Biology, Cambridge, UK.
⁴ Department of Basic and Clinical Neuroscience, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London SE5 9RT, UK; MRC Centre for Neurodevelopmental Disorders, King's College London, London, UK; Dementia Research Institute, King's College London, London, UK. Electronic address: deepak.srivastava@kcl.ac.uk.

PMID: 41903542
DOI: 10.1016/j.crmeth.2026.101371

Abstract

Microelectrode arrays (MEAs) are increasingly used to profile the development of synchronized activity in neural organoids, yet no organoid study has reported on the consistency of electrophysiological development across cell lines. Here, we used dissociated neural organoids derived from six cell lines on MEAs to characterize functional synapse development using multiple parameters across time. The dissociated organoids demonstrated 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. These results demonstrate that dissociated cerebral organoids can generate functional neurons, akin to primary neuronal cultures from brain tissue, providing a scalable model for studies of neurodevelopment and synaptic function. Consistent with unguided differentiation, we observed variability in activity parameters linked to donor cell line and batch effects, which must be considered in experimental design.

Keywords: CP: stem cell; MEA; cerebral organoids; electrophysiology; estradiol; functional connectivity; human induced pluripotent stem cells; micro-electrode arrays; network dynamics; neuronal cultures; synaptic plasticity.