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Front. Neurosci. | doi: 10.3389/fnins.2018.00094

Simple and inexpensive paper-based astrocyte co-cultures to improve survival of low-density neuronal networks

 Mathias J. Aebersold1,  Greta Thompson-Steckel1,  Adriane Joutang1,  Moritz Schneider1, Conrad Burchert1, Csaba Forró1, Serge Weydert1, Hana Han1 and  János Vörös1*
  • 1Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, Switzerland

Bottom-up neuroscience aims to engineer well-defined networks of neurons to investigate the functions of the brain. By reducing the complexity of the brain to achievable target questions, such in vitro bioassays better control experimental variables and can serve as a versatile tool for fundamental and pharmacological research. Astrocytes are a cell type critical to neuronal function, and the addition of astrocytes to neuron cultures can improve the quality of in vitro assays. Here, we present cellulose as an astrocyte culture substrate. Astrocytes cultured on the cellulose fiber matrix thrived and formed a dense 3D network. We devised a novel co-culture platform by suspending the easy-to-handle astrocytic paper cultures above neuronal networks of low densities typically needed for bottom-up neuroscience. There was drastic improvement in neuronal viability after 5 days in vitro at densities ranging from 50,000 cells/cm2 down to isolated cells at 1,000 cells/cm2. Cultures exhibited spontaneous spiking even at the very low densities, with a significantly greater spike frequency per cell compared to control mono-cultures. Applying the co-culture platform to an engineered network of neurons on a patterned substrate resulted in significantly improved viability and almost doubled the density of live cells. Lastly, the shape of the cellulose substrate can easily be customized to a wide range of culture vessels, making the platform versatile for different applications that will further enable research in bottom-up neuroscience and drug development.

Keywords: Neuron, astrocyte, co-culture, Paper-based, Low-density culture, Cell viability, Neurite length, network activity

Received: 06 Nov 2017; Accepted: 05 Feb 2018.

Edited by:

Sergio Martinoia, Università di Genova, Italy

Reviewed by:

Bruce C. Wheeler, University of Florida, United States
Andreas Offenhäusser, Forschungszentrum Jülich, Germany  

Copyright: © 2018 Aebersold, Thompson-Steckel, Joutang, Schneider, Burchert, Forró, Weydert, Han and Vörös. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Prof. János Vörös, ETH Zurich, Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, Zurich, Switzerland,