Synaptic Microcircuit Modeling with 3D Cocultures of Astrocytes and Neurons from Human Pluripotent Stem Cells

This method can help answer key questions in neuroscience regarding how intercellular communication between multiple human neural cell types contributes to synaptic circuit formation and function. This technique rapidly and systematically generates three-dimensional coculture spheres of functionally-mature astrocytes and neurons from human pluripotent stem cells. These can be utilized for rigorous experimental investigation.

The reproducibility and scalability of this protocol is a defined alternative to the emerging organoid technologies. These show promise for novel drug development and cellular-engraftment therapy to promote neuroregeneration after injury or disease. Begin by seating clusters of HPSCs in two milliliters of HPSC medium containing the rho-kinase inhibitor Y-27632 into each well of an ECM-coated six-well plate.