All-defined NGN2 neuron protocol reveals multifaceted signatures of neuronal maturation [bulk_media]
Direct conversion of iPSCs into neurons by NGN2 over-expression is a popular method for disease modeling. Here we report a standardized method to generate functional excitatory cortical neurons using clonal, targeted engineered iPSC lines and defined reagents. We demonstrate that our protocol generates consistent excitatory cortical neurons at scale. These cultures can be maintained healthily for at least 150 days and expresses the majority of the GWAS genes found in autism, Parkinson's disease, and Alzheimer's disease. Comprehensive temporal omics, functional, and morphological profiling demonstrated continued progression of neuronal maturation. Deep functional characterization through transcriptomic, imaging, and functional assays revealed the coordinated action of multiple pathways in neuronal maturation. Our method and profiling data can serve as reference for developing human in vitro neuronal models for disease modeling. Overall design: Bulk RNA-seq of iPSC-derived NGN2-induced neurons. Neurons were cultured in either home brew media or mouse astrocyte-conditioned media and collected at differentiation day 42. For additional details, please refer to Shan et al.