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Structuring a multi-nodal neural network in vitro within a novel design microfluidic chip

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Abstract

Neural network formation is a complex process involving axon outgrowth and guidance. Axon guidance is facilitated by structural and molecular cues from the surrounding microenvironment. Micro-fabrication techniques can be employed to produce microfluidic chips with a highly controlled microenvironment for neural cells enabling longitudinal studies of complex processes associated with network formation. In this work, we demonstrate a novel open microfluidic chip design that encompasses a freely variable number of nodes interconnected by axon-permissible tunnels, enabling structuring of multi-nodal neural networks in vitro. The chip employs a partially open design to allow high level of control and reproducibility of cell seeding, while reducing shear stress on the cells. We show that by culturing dorsal root ganglion cells (DRGs) in our microfluidic chip, we were able to structure a neural network in vitro. These neurons were compartmentalized within six nodes interconnected through axon growth tunnels. Furthermore, we demonstrate the additional benefit of open top design by establishing a 3D neural culture in matrigel and a neuronal aggregate 3D culture within the chips. In conclusion, our results demonstrate a novel microfluidic chip design applicable to structuring complex neural networks in vitro, thus providing a versatile, highly relevant platform for the study of neural network dynamics applicable to developmental and regenerative neuroscience.

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Acknowledgements

This work was supported by the Research Council of Norway, Norwegian Micro- and Nano-Fabrication Facility, NorFab, project number 245963/F50, NTNU program for Enabling Technologies, (Nanotechnology), and the Liaison Committee between the Central Norway Health Authority and NTNU (Samarbeidsorganet HMN-NTNU).

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Authors and Affiliations

  1. Department of Clinical and Molecular Medicine, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, PO Box 8905 MTFS, NO-7491, Trondheim, Norway

    Rosanne van de Wijdeven & Øyvind Halaas

  2. Department of Neuromedicine and Movement Science, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, N-7491, Trondheim, Norway

    Ola Huse Ramstad, Ulrich Stefan Bauer, Axel Sandvig & Ioanna Sandvig

  3. Department of Pharmacology and Clinical Neurosciences, Division of Neuro, Head and Neck, Umeå University Hospital, SE-90187, Umeå, Sweden

    Axel Sandvig

  4. Department of Neurology and Clinical Neurophysiology, St Olavs Hospital, 7030, Trondheim, Norway

    Axel Sandvig

Authors
  1. Rosanne van de Wijdeven
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  2. Ola Huse Ramstad
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  3. Ulrich Stefan Bauer
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  4. Øyvind Halaas
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  5. Axel Sandvig
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  6. Ioanna Sandvig
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Corresponding author

Correspondence to Rosanne van de Wijdeven.

Ethics declarations

All animal procedures were in accordance with the EU Directive 86/609/EEC and the Norwegian laws and regulations controlling procedures on experimental animals.

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There are no conflicts to declare.

Additional information

Rosanne van de Wijdeven and Ola Huse Ramstad share first authorship

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van de Wijdeven, R., Ramstad, O.H., Bauer, U.S. et al. Structuring a multi-nodal neural network in vitro within a novel design microfluidic chip. Biomed Microdevices 20, 9 (2018). https://doi.org/10.1007/s10544-017-0254-4

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  • DOI: https://doi.org/10.1007/s10544-017-0254-4

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