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Synaptic Microcircuit Modeling with 3D Cocultures of Astrocytes and Neurons from Human Pluripotent Stem Cells
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Chapters
- 00:04Title
- 00:49Generation and Maintenance of Astrocyte Progenitors (hAstros) from hPSCs
- 02:41Generation and Maintenance of Neurons (hNeurons) and Inducible Neurons (iNeurons) from hPSCs
- 03:27Preparation and Maintenance of 3D Sphere Co-cultures
- 05:42Measurement of Live Synaptic Physiology with Multielectrode Arrays (MEAs)
- 06:56Results: Morphological and Physiological Characteristics of Synaptic Microcircuits
- 08:07Conclusion
In this protocol, we aim to describe a reproducible method for combining dissociated human pluripotent stem cell derived neurons and astrocytes together into 3D sphere cocultures, maintaining these spheres in free floating conditions, and subsequently measuring synaptic circuit activity of the spheres with immunoanalysis and multielectrode array recordings.
Tags
Keywords: Synaptic Microcircuit3D CoculturesAstrocytesNeuronsHuman Pluripotent Stem CellsNeural Cell TypesSynaptic Circuit FormationNeuroscienceOrganoid TechnologiesDrug DevelopmentNeuroregenerationNeural ProgenitorsAstrocyte ProgenitorsAstrocyte SubtypesNeural InductionNeural MediumRho-kinase InhibitorECM-coated PlatesCell AggregationCell DissociationCell Identity ConfirmationCryo-preservation
