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University of Genova

3 ARTICLES PUBLISHED IN JoVE

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 Bonifazi^1,4

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.

Neuroscience

Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model

Mariateresa Tedesco¹, Monica Frega^1,2, 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.

Developmental Biology

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

Monica Frega^*1,2, Sebastianus H. C. van Gestel^*3, Katrin Linda^2,3, Jori van der Raadt³, Jason Keller^1,2, Jon-Ruben Van Rhijn^1,2, Dirk Schubert^1,2, Cornelis A. Albers^2,3,4, Nael Nadif Kasri^1,2,3

¹Department of Cognitive Neurosciences, Radboudumc, ²Donders Institute for Brain, Cognition and Behaviour, Radboud University, ³Department of Human Genetics, Radboudumc, ⁴Department of Molecular Developmental Biology, Radboud University

We modify and implement a previously published protocol describing the rapid, reproducible, and efficient differentiation of human induced Pluripotent Stem Cells (hiPSCs) into excitatory cortical neurons¹². Specifically, our modification allows for control of neuronal cell density and use on micro-electrode arrays to measure electrophysiological properties at the network level.