Phenotypic assay development with iPSC-derived neurons

Induced pluripotent stem cell (iPSC)-derived neurons have been identified as a promising model system to improve translation of preclinical neuroscience studies to the clinic. The use of patient-derived neurons to study neurodevelopmental disorders enhances confidence in target and pathway identification because the neurons express disease-causing variants and genetic backgrounds that are unique to patients diagnosed with these disorders. There are many phenotypic assays that can be used to understand how genetic or pharmacological manipulation of iPSC-derived neurons affects neuron identity and function, including high content imaging and functional recording of neuronal activity. This chapter discusses some of the technical considerations required to successfully collect and interpret phenotypic data using iPSC-derived neurons. These considerations include the differentiation protocol chosen, coating materials, plating conditions, and equipment options, among others. Although this chapter is not exhaustive, it will provide a framework for any lab starting work using iPSC-derived neurons to model neurological disorders.