Abstract
Generation of neuronal cultures from induced pluripotent stem cells (hiPSCs) serve the studies of human brain disorders. However we lack neuronal networks with balanced excitatory-inhibitory activities, which are suitable for single cell analysis. We generated low-density networks of hPSC-derived GABAergic and glutamatergic cortical neurons. We used two different co-culture models with astrocytes. We show that these cultures have balanced excitatory-inhibitory synaptic identities using confocal microscopy, electrophysiological recordings, calcium imaging and mRNA analysis. These simple and robust protocols offer the opportunity for single-cell to multi-level analysis of patient hiPSC-derived cortical excitatory-inhibitory networks; thereby creating advanced tools to study disease mechanisms underlying neurodevelopmental disorders.
Authors
Aishwarya G Nadadhur, Javier Emperador Melero, Marieke Meijer, Desiree Schut, Gerbren Jacobs, Ka Wan Li, J J Johannes Hjorth, Rhiannon M Meredith, Ruud F Toonen, Ronald E Van Kesteren, August B Smit, Matthijs Verhage, Vivi M Heine