To speed up the development of more effective and safer drugs, there is an increasing need for more complex, biologically relevant, and predictive cell-based assays for drug discovery and toxicology screening. Because the human Central Nervous System (CNS) is a structurally and functionally complex organ, creating an accurate and predictive in-vitro model has been challenging. StemoniX has commercialized innovative organoid technology with microBrain® 3D: pre-plated neural spheroids composed of human iPSC-derived cortical neurons and astrocytes of uniform composition and size.
This tutorial will detail how the FLIPR Penta® System and ImageXpress® Micro Confocal System (Molecular Devices) have been used to functionally and structurally characterize microBrain 3D spheroids from both healthy and disease sources. We describe a method for the complex analysis of calcium oscillations that allows detection and multi-parametric characterization of neuronal activity. In addition, cellular and mitochondrial toxicity were assessed by high-content imaging. The complementary approaches have allowed a thorough characterization of microBrain 3D neural spheroids and demonstrates how it can be used across drug discovery for disease modeling, screening, and toxicity testing.