Petersilie et al. 2023 - Figure 2
Loading...
Date
2023
Journal Title
Journal ISSN
Volume Title
Publisher
iScience
Abstract
Brain organoids derived from human pluripotent stem cells (hPSCs) are a promising
tool for studying human neurodevelopment and related disorders. Here, we generated
long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid
interphase from regionalized cortical organoids. We demonstrate that cBOS host
mature neurons and astrocytes organized in complex architecture. By leveraging the
fact that cBOS allow direct access to the developing neural cells, we carried out an
array of functional analyses. Whole-cell patch-clamp demonstrated subthreshold
synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium
signals turned into synchronous large-scale calcium oscillations upon combined
disinhibition of NMDA receptors and blocking of GABAA receptors. Lastly, FRET-based
imaging with the genetically-encoded nanosensor ATeam1.03YEMK revealed the high
sensitivity of neurons to acute metabolic inhibition. Altogether, cBOS represent a
powerful platform for assessing the morphological and functional aspects of organized
human neural cells in intact minimal networks.
Description
Keywords
Brain organoids, pluripotent stem cells, neurons, astrocytes, synaptic protein expression, electrophysiology, calcium signaling, glutamate, GABA, ATP