Current methods differentiate cerebral cortex organoids from stem cells resulting in a high heterogeneity of cells that yields a high variability in results.

A University of Utah researcher has developed a method for generating three-dimensional cortical organoids from single induced pluripotent stem cell (iPSC)-derived neural rosettes in vitro. Single-rosette-derived cortical organoids grow large in suspension culture, reaching 4-5 mm in diameter by 4 months while maintaining a single internal lumen. They consist of different types of neuronal cells, including cortical neural progenitors, deep and superficial cortical excitatory neurons, inhibitory neurons, and astrocytes, organized around the lumen. Patch-clamp electrophysiology shows that many neurons in slices from single-rosette-derived cortical organoids fire repetitive action potentials, receive excitatory and inhibitory synaptic inputs, exhibit typical pyramidal-like morphologies, and develop dendritic spines. These organoids are useful for defining the organization and functions of human neurons in healthy and diseased developing cortical networks and have potential applications in regenerative medicine and transplantation such as treating certain forms of drug-resistant epilepsy.