University of Genova 2 articles published in JoVE Neuroscience Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model Mariateresa Tedesco1, Monica Frega1,2, Sergio Martinoia1, Mattia Pesce3, Paolo Massobrio1 1Department of Informatics, Bioengineering, Robotics and System Engineering (DIBRIS), University of Genova, 2Donders Institute for Brain, Cognition and Behaviour, Department of Cognitive Neuroscience, Radboud University Medical Center, 3Fondazione Istituto Italiano di Tecnologia (IIT) In this work, a novel experimental model in which 3D neuronal cultures are coupled to planar Micro-Electrode Arrays (MEAs) is presented. 3D networks are built by seeding neurons in a scaffold made up of glass microbeads on which neurons grow and form interconnected 3D structures. Neuroscience Design, Surface Treatment, Cellular Plating, and Culturing of Modular Neuronal Networks Composed of Functionally Inter-connected Circuits Sivan Kanner*1, Marta Bisio*2, Gilad Cohen3, Miri Goldin4, Marieteresa Tedesco5, Yael Hanein3, Eshel Ben-Jacob4, Ari Barzilai1, Michela Chiappalone2, Paolo Bonifazi1,4 1Department of Neurobiology, George S. Wise Faculty of Life Sciences, Tel-Aviv University, 2Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, 3School of Electrical Engineering, Tel-Aviv University, 4School of Physics and Astronomy, Tel-Aviv University, 5Department of Informatics, Bioengineering, Robotics and System Engineering, University of Genova This manuscript describes a protocol to grow in vitro modular networks consisting of spatially confined, functionally inter-connected neuronal circuits. A polymeric mask is used to pattern a protein layer to promote cellular adhesion over the culturing substrate. Plated neurons grow on coated areas establishing spontaneous connections and exhibiting electrophysiological activity.