Tuberous sclerosis complex (TSC) is a rare neurodevelopmental disorder resulting from autosomal dominant mutations in the TSC1 or TSC2 genes, leading to a hyperactivated mammalian target of rapamycin (mTOR) pathway, and gray and white matter defects in the brain. To study the involvement of neuron-glia interactions in TSC phenotypes, we generated TSC patient induced pluripotent stem cell (iPSC)-derived cortical neuronal and oligodendrocyte (OL) cultures. TSC neuron mono-cultures showed increased network activity, as measured by calcium transients and action potential firing, and increased dendritic branching. However, in co-cultures with OLs, neuronal defects became more apparent, showing cellular hypertrophy and increased axonal density. In addition, TSC neuron-OL co-cultures showed increased OL cell proliferation and decreased OL maturation. Pharmacological intervention with the mTOR regulator rapamycin suppressed these defects. Our patient iPSC-based model, therefore, shows a complex cellular TSC phenotype arising from the interaction of neuronal and glial cells and provides a platform for TSC disease modeling and drug development. •TSC neuron mono-cultures show an increase in network activity and dendritic branching•TSC co-cultures show hypertrophy and an increase in axonal length and OL proliferation•mTOR regulators normalize TSC neuronal and glial phenotypes Nadadhur et al. generated TSC disease models using patient iPSCs. While neuron mono-cultures showed an increase in network activity and dendritic branching, only when co-cultured with oligodendrocytes (OLs), hypertrophy and axonal abnormalities were observed. Neuron-OL interactions, modulated by mTOR regulators, support use of mixed cultures for TSC disease modeling and drug development.